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Long-fermentation doughs: role of gluten structure and differences between strong flours and einkorn flours

by luciano

Highlights

Long fermentations do not depend exclusively on flour “strength”
The idea that only strong flours are suitable is an operational simplification that does not capture the complexity of the gluten system.

Gluten is a dynamic system, not a static one
The gluten network forms and evolves over time through continuous processes of bond breaking and reorganization.

Dough stability depends on the continuity of the protein network
It is not only about “how much gluten,” but how it is organized into a connected three-dimensional structure.

There is a critical threshold of structural collapse
When network continuity is lost, the dough rapidly shifts from stable to unstable, with non-linear behavior.

Long fermentations modify the gluten network
Through:

  • proteolysis

  • thiol–disulfide exchange

  • changes in redox state

Strong and weak flours differ in their distance from the critical threshold

  • strong flours → more extended and stable network

  • weak flours → more fragile network, closer to collapse

Einkorn represents a limiting model

  • less organized and less elastic network

  • higher sensitivity to degradation

  • more plastic behavior

Collapse can be reversible or irreversible

  • elastic → recoverable

  • plastic → permanent loss of structure

Dough recovery is a reorganization, not a “reactivation”
Proteins do not regenerate: they restructure, temporarily increasing connectivity.

Key practical implication
Managing long fermentations requires control of network structural continuity, not just flour selection.

1️⃣ Introduction

In baking practice, it is widely believed that long fermentations necessarily require strong flours. Although this indication is often useful operationally, it does not account for the structural and dynamic nature of gluten.

“The quality of a dough does not depend exclusively on the quantity of proteins, but on their organization into a three-dimensional viscoelastic network, whose stability evolves over time under the influence of enzymatic and physicochemical phenomena (Wieser, 2007).”

2️⃣ Gluten as a dynamic system

Gluten is not a pre-formed structure, but a system that emerges during hydration and mixing. It is mainly composed of:

gliadins → provide extensibility
glutenins → provide elasticity
a very high molecular weight fraction called GMP (Glutenin Macropolymer) → the elastic backbone of the dough

GMP represents the fundamental structural component for the formation of a continuous network capable of retaining gas (Don et al., 2005).

The behavior of gluten is intrinsically dynamic: the protein network is subject to continuous processes of bond breaking and reformation, particularly disulfide bonds and non-covalent interactions (Wieser, 2007; Belton, 1999).

The gluten network progressively organizes into a three-dimensional matrix capable of retaining gas and water during mixing and fermentation. In this context, non-protein components such as arabinoxylans can also physically interact with the matrix, creating a secondary network that may strengthen the structure or, in some cases, hinder protein aggregation (Courtin & Delcour, 2002).

3️⃣ Evolution of the network during long fermentations

During long fermentations, three main phenomena are observed:

  1. Proteolysis: endogenous and microbial enzymes reduce the length of protein chains (Thiele et al., 2002)

  2. Thiol–disulfide exchange: covalent bonds between proteins are continuously reorganized

  3. Changes in redox state: metabolites produced by microorganisms influence the oxidation–reduction balance (Grosch & Wieser, 1999)

These processes lead to a progressive modification of the connectivity of the gluten network.

4️⃣ The critical threshold of structural collapse

Dough stability can be interpreted in terms of continuity of the protein network. As long as a connected structure spans the entire system, the dough maintains its mechanical properties.

Below a certain critical threshold, this continuity is lost and the system collapses. This behavior is consistent with percolation models of polymer networks, in which emergent properties depend on the global connectivity of the system (Stauffer & Aharony, 1994).

As a result, the transition from a stable to an unstable state can occur suddenly and non-linearly.

5️⃣ Elastic collapse vs plastic collapse

From a rheological point of view, it is useful to distinguish between:

Elastic collapse (reversible)
soft but cohesive dough
ability to recover through mechanical handling
network still continuous but relaxed

Plastic collapse (irreversible)
incoherent and sticky dough
loss of gas retention capacity
absence of response to deformation

This distinction is consistent with rheological models of dough, which highlight the transition from viscoelastic to plastic behavior (Dobraszczyk & Morgenstern, 2003).

6️⃣ Strong flour vs weak flour: it’s not just “how much gluten”

The difference between flours does not lie solely in total protein content, but in structural parameters such as:

distribution of glutenin molecular weights
content of high molecular weight subunits
initial GMP density
stability of disulfide bonds
gliadin/glutenin ratio

Strong flours exhibit a more extended and stable initial network, placing them further from the critical collapse threshold. Weak flours, on the contrary, operate closer to this threshold and are therefore more sensitive to proteolysis and environmental variations (MacRitchie, 1999; Payne, 1987).

7️⃣ The case of einkorn (Triticum monococcum)

Einkorn represents a particularly useful system for analyzing dough behavior under conditions close to the critical threshold of structural collapse.

Compared to modern wheats, it is characterized by:

lower capacity to form Glutenin Macropolymer (GMP)
reduced presence of high molecular weight glutenin subunits
less extended and less elastic protein network
more plastic rheological behavior

These characteristics result in an intrinsically less stable structure, placing the dough near the critical threshold of continuity (Hidalgo & Brandolini, 2014).

Under long fermentation conditions, this configuration makes the system particularly sensitive to proteolytic phenomena and changes in redox state. As a result, the dough may show a marked loss of consistency, appearing macroscopically collapsed.

However, this state does not necessarily imply that the critical threshold has been exceeded.

If a continuous network is still present, even if highly weakened, moderate mechanical interventions can induce a reorganization of the structure, with partial recovery of rheological properties.

This behavior can be interpreted as a reorganization of the protein network, made possible by:

realignment of protein chains
reorganization of thiol–disulfide bonds
increase in local connectivity
partial restructuring of GMP

The observed recovery does not correspond to an increase in the intrinsic “strength” of the flour, but to a temporary restoration of structural continuity.

In this sense, einkorn constitutes an effective experimental model for making visible structural transition phenomena that are less evident in stronger flours.

Experimental case – structural recovery after long fermentation

In a test conducted on einkorn dough:

total maturation: 36 hours
12 hours at 18 °C
24 hours at 5 °C
subsequent rest at room temperature: 1–2 hours

The dough initially appeared highly degraded, with apparent loss of structure and behavior comparable to collapse.

However, the application of two light folds led to a significant recovery of shape and cohesion.

This result suggests that the system had not exceeded the critical threshold of structural continuity.

Mechanical handling likely promoted a reorganization of the protein network, temporarily increasing internal connectivity.

The described test is taken from: “Advanced methodology for producing bread doughs with flours of limited gluten development capacity”, available at www.glutenlight.eu

Scientific reference
Hidalgo, A., Brandolini, A. (2014). Nutritional properties of einkorn wheat. DOI: 10.1016/j.jcs.2014.04.005
Summary: einkorn exhibits a less organized protein structure and lower baking quality compared to modern wheats.

Complementary reference
Brandolini, A., Hidalgo, A. (2011). Einkorn wheat: a review
Summary: less elastic doughs, greater structural fragility, more plastic behavior.

8️⃣ Reorganization vs “reactivation”

It is important to distinguish between biological and physical phenomena. Gluten proteins do not regenerate or become “reactivated.”

The recovery observed in some doughs is attributable to a reorganization of the protein network, in which fractions initially not integrated into the GMP can progressively become involved (Belton, 1999).

A portion of proteins initially not fully integrated into the GMP may progressively be incorporated during:

maturation
handling
rest

This can temporarily increase network continuity. It is a physical phenomenon, not a biological one.


9️⃣ Practical implications

From a practical standpoint, evaluating dough condition can be reduced to its structural continuity:

cohesive but relaxed dough → potentially recoverable
incoherent dough → likely beyond the critical threshold

Mechanical operations (e.g., folds) are effective only if a continuous network is still present.

Conclusions

Long fermentations do not necessarily require strong flours, but rather careful management of the parameters that regulate gluten network stability:

control of proteolysis
temperature management
redox state regulation
respect of the critical continuity threshold

The difference between strong and weak flours is mainly quantitative: the former operate far from the collapse threshold, the latter close to it. Under these conditions, system sensitivity becomes the determining factor.

Further insights

1 – Arabinoxylans
Non-starch polysaccharides that interact with the gluten matrix, influencing hydration, viscosity, and continuity of the protein network.
→ Further reading: Arabinoxylans

2 – Dough redox state
The oxidation–reduction balance regulates the formation and reorganization of disulfide bonds, directly affecting gluten network stability during fermentation.
→ Further reading: Redox

Essential references

  • Belton, P.S. (1999). On the elasticity of wheat gluten. DOI: 10.1098/rstb.1999.0414

  • Courtin, C.M., Delcour, J.A. (2002). Arabinoxylans and endoxylanases in wheat flour bread-making. DOI: 10.1016/S0733-5210(02)00011-1

  • Dobraszczyk, B.J., Morgenstern, M.P. (2003). Rheology and the breadmaking process. DOI: 10.1016/S0268-005X(03)00059-6

  • Don, C. et al. (2005). Glutenin macropolymer and dough properties. DOI: 10.1016/j.jcs.2004.07.001

  • Hidalgo, A., Brandolini, A. (2014). Nutritional properties of einkorn wheat. DOI: 10.1016/j.jcs.2014.04.005

  • MacRitchie, F. (1999). Wheat proteins and functionality

  • Payne, P.I. (1987). Genetics of wheat storage proteins

  • Shewry, P.R., Halford, N.G. (2002). DOI: 10.1093/jxb/53.370.947

  • Stauffer, D., Aharony, A. (1994). Introduction to Percolation Theory

  • Thiele, C. et al. (2002). DOI: 10.1128/AEM.68.3.1206-1213.2002

  • Wieser, H. (2007). DOI: 10.1016/j.foodmicro.2006.07.004

Scientific Evidence and Application Limits Chapter IV

by luciano

1. Scope and operational definitions

In technical language it is essential to separate three concepts that are often confused:

1. Gluten hydrolysis/proteolysis
→ fragmentation of proteins (gliadins and glutenins) into smaller peptides.

  1. Reduction of immunogenic peptides/epitopes for celiac disease→ degradation of specific sequences rich in proline and glutamine (e.g. “Pro-rich” peptides) that resist digestion and activate immune responses in celiac patients.

  1. “Elimination” of gluten→ a much more ambitious objective, achievable only under controlled technological conditions (selected strains, often enzymatic co-adjuvants, long fermentation times), and not equivalent to normal baking with traditional sourdough.

2. Evidence: what studies show

2.1 Fermentation with selected lactic acid bacteria: targeted degradation of immunogenic peptides

Di Cagno et al., 2004 (Applied and Environmental Microbiology) demonstrate that the use of selected lactobacilli with specialized peptidases is able to hydrolyze proline-rich peptides, including peptides with high immunogenicity (the work explicitly discusses the hydrolysis of “Pro-rich” peptides and the application to an experimental baked product).
The study also includes an acute clinical challenge test in subjects with celiac disease within the described experimental protocol. (PubMed)

Key technical points (what is “demonstrated”)

  • The ability to degrade prolamin fractions critically depends on strain selection (it is not an automatic effect of any sourdough). (PubMed)

  • Degradation involves peptides known to resist gastrointestinal digestion thanks to enzymatic systems (peptidases) not typical of baker’s yeast alone. (PubMed)

Immediate applicative limit

The protocol is not “generic sourdough”: it is a biotechnology using selected strains and defined conditions; it is not automatically transferable to any artisanal process. (PubMed)

2.2 “Enhanced” fermentation: selected lactobacilli + fungal proteases (extensive detoxification)

Rizzello et al., 2007 (Applied and Environmental Microbiology) show an even more “engineered” approach: a mixture of selected lactobacilli + fungal proteases during prolonged fermentation.

The study uses several analytical techniques (immunological and instrumental) to estimate residual gluten and the persistence of different protein fractions. (PubMed)

Key technical points

  • Complete hydrolysis of gliadins and other soluble fractions reported in the experimental process; partial persistence of a fraction of glutenins (not all structural fractions are necessarily “eliminated”). (PubMed)

  • Measurement of residual gluten through immunological tests (R5-ELISA) and confirmation through proteomic/spectrometric analyses in the protocol. (PubMed)

  • Biological evaluation of immunoreactivity (tests on immune cell lines) to estimate the “toxicity” of the pepsin-trypsin digest of the fermented product. (PubMed)

Applicative limit

This scenario requires enzymatic co-adjuvants (fungal proteases) and a controlled setup: it is an industrial/biotechnological process, not the equivalent of standard sourdough management in a bakery. (PubMed)

2.3 Selected lactic fermentation on different cereals: role of pH and endogenous enzymes

De Angelis et al., 2006 (Journal of Cereal Science) study the fermentation of rye flours with selected lactic acid bacteria, showing extensive hydrolysis of ethanol-soluble polypeptides and a reduction of immunochemical detectability (R5-Western), also discussing the role of pH in activating hydrolysis through endogenous flour enzymes. (ScienceDirect)

Key technical points

The observed degradation results from a combination of:

  • microbial proteolytic activity (selected strains)

  • pH-dependent hydrolysis (activation of endogenous cereal enzyme systems) (ScienceDirect)

The work supports the “biotechnological” logic of controlled fermentation as a tool to reduce contamination/reactivity risk in specific contexts (in experimental terms). (ScienceDirect)

Applicative limit

Again: selected strains + defined process conditions; this is not an automatic generalization for “any sourdough.” (ScienceDirect)

3. Where baker’s yeast and “traditional” sourdough fit

3.1 Baker’s yeast (Saccharomyces cerevisiae)

Within the framework of the above studies, the effect of baker’s yeast is mainly:

  • fermentative kinetics (CO₂, volumetric development)

  • indirect influence on maturation (time/temperature)

but not a proteolytic activity comparable to that of selected lactic bacteria and/or added proteases.

In other words: with baker’s yeast the “improved digestive management” (when observed) is more related to maturation time and transformations of the starch-protein matrix, not to extensive degradation of immunogenic gluten sequences (in the terms used in the cited studies).

(This is a conclusion derived by comparing the mechanisms reported in studies on selected LAB and proteases.) (PubMed)

3.2 “Non-selected” sourdough (spontaneous sourdough starter)

Spontaneous sourdough can determine:

  • acidification

  • partial proteolysis

  • rheological modifications

However, the literature showing “almost total” degradation or marked reduction of immunogenic epitopes typically uses:

  • selected lactic strains with specific peptidases (PubMed)

and/or

  • fungal proteases in combination (PubMed)

Therefore, at a manualistic level, the correct formulation is:

Fermentation with sourdough can increase gluten proteolysis; extensive degradation of immunogenic sequences requires controlled biotechnological protocols (selected strains and, in some cases, enzymatic co-adjuvants).

4. Applicative limits

Protocols aiming to drastically reduce the immunogenic fraction of gluten do not coincide with the standard production of sourdough bread/pizza. (PubMed)

The result depends on:

  • microbial species/strains used (selection) (PubMed)

  • fermentation time

  • acidity/pH (and relative enzymatic activation) (ScienceDirect)

  • possible use of technological proteases (PubMed)

Even when very extensive degradation is observed, some studies report possible persistence of certain fractions (e.g. part of the glutenins) depending on the protocol. (PubMed)

5. Cited studies

Di Cagno, R. et al. (2004). Sourdough bread made from wheat and nontoxic flours and started with selected lactobacilli is tolerated in celiac sprue patients. Applied and Environmental Microbiology, 70(2), 1088–1096. DOI: 10.1128/AEM.70.2.1088-1096.2004 (PubMed)

Rizzello, C.G. et al. (2007). Highly efficient gluten degradation by lactobacilli and fungal proteases during food processing: new perspectives for celiac disease. Applied and Environmental Microbiology, 73(14), 4499–4507. DOI: 10.1128/AEM.00260-07 (PubMed)

De Angelis, M. et al. (2006). Fermentation by selected sourdough lactic acid bacteria to decrease coeliac intolerance to rye flour. Journal of Cereal Science, 43(3), 301–314. DOI: 10.1016/j.jcs.2005.12.008 (ScienceDirect)

In-depth analysis

Effects of sourdough and/or yeast use in gluten fermentation: scientific evidence

Primary studies (main evidence)

1. Effects of LAB + yeast co-fermentation on gluten degradation

Title: Effects of Co-Fermentation with Lactic Acid Bacteria and Yeast on Gliadin Degradation in Whole-Wheat Sourdough

Summary: The study evaluates how selected strains of Lactic Acid Bacteria (LAB) and baker’s yeast (Saccharomyces cerevisiae) co-ferment gluten in whole-wheat sourdough. The combined fermentation leads to significant degradation of gliadin and glutenin fractions, with reduction of gluten content. Strains such as Lactobacillus brevis and Pediococcus pentosaceus show high proteolytic activity. (MDPI)

2. Reduction of gluten allergenicity in fermented products

Title: From gluten structure to immunogenicity: Investigating the effects of lactic acid bacteria and yeast co-fermentation on wheat allergenicity in steamed buns

Summary: LAB + baker’s yeast co-fermentation induces depolymerization of gluten macromolecules and reduces total immunoreactivity compared with non-fermented controls. Significant decreases in α/γ-gliadins and glutenins associated with celiac disease are observed. (PubMed)

3. Immunogenic peptides and sourdough

Title: A Case Study of the Response of Immunogenic Gluten Peptides to Sourdough Proteolysis

Summary: Fermentation with sourdough modifies gluten structure and the release profile of immunogenic peptides during in vitro digestion, without necessarily eliminating them completely. Comparative study between sourdough bread and rapid-leavened bread. (PubMed)

4. Bacillus spp. isolated from sourdough and gluten hydrolysis

Title: Gluten hydrolyzing activity of Bacillus spp isolated from sourdough

Summary: Bacillus strains isolated from sourdough degrade the immunogenic 33-mer peptide and gliadin sequences, reducing gluten below 110 mg/kg. Potential application in reduced-gluten products. (SpringerLink)

5. Pilot clinical study on fermented products

Title: Gluten-free sourdough wheat baked goods appear safe for young celiac patients: a pilot study

Summary: Fermentation with selected lactobacilli and fungal proteases reduces gluten below 10 ppm. Products tested on children with celiac disease in remission show good clinical tolerability. (PubMed)

6. Recent review on the role of fermentation (2025)

Title: Sourdough Fermentation and Gluten Reduction: A Biotechnological Approach for Gluten-Related Disorders

Summary: LAB fermentation contributes to the reduction of gluten peptides but is not sufficient alone to eliminate all immunogenic sequences. Combined processes with exogenous proteases are more effective. (MDPI)

Previously cited studies, with greater detail

A. Bacillus spp isolated from sourdough
DOI: 10.1186/s12934-020-01388-z

Further detail: The study demonstrates the high proteolytic activity of Bacillus strains against gliadin substrates and the 33-mer peptide. Extensive hydrolysis leads to gluten levels <110 mg/kg in fermented sourdough.

B. Label-free quantitative proteomics and sourdough fermentation
DOI: 10.1016/j.foodchem.2023.137037

Further detail: Proteomic analysis identifies 85 allergenic proteins modulated by fermentation. Some microbial combinations show reduction of gliadins containing immunogenic sequences, suggesting a selective effect of fermentation on the wheat protein fraction.

C. Yeast–bacteria interactions and immunogenicity
DOI: 10.1016/j.ifset.2023.103281

Further detail: Co-cultures of yeasts (Saccharomyces, Torulaspora) with Pediococcus acidilactici show greater gluten depolymerization and reduced immunogenicity compared with single-yeast fermentations.

General conclusions

Sourdough fermentation can partially degrade gluten and reduce specific immunogenic peptides. The reduction does not equal complete elimination: without exogenous proteases, residual gluten often remains. Effectiveness strongly depends on microbial strains and fermentation conditions.

What does all this mean for those seeking gluten-light products?

Products made with sourdough (sourdough fermentation) generally present technological and biochemical characteristics superior to products obtained with rapid leavening, especially regarding tolerability and overall quality.

In particular:

Partial gluten degradation

Prolonged fermentation promotes hydrolysis of some gliadin and glutenin fractions, reducing protein complexity compared with non-fermented doughs.

Modified peptide profile

Even when gluten is not eliminated, its structure changes, with a potential reduction of specific immunogenic peptides.

Perceived improved digestibility

Many non-celiac consumers report better gastrointestinal tolerance compared with industrial baked products produced with rapid fermentation.

Reduction of other critical factors

Sourdough fermentation also contributes to decreasing FODMAPs and some antinutritional compounds.

⚠️ Important note: gluten-light products are not automatically safe for people with celiac disease. Traditional fermentation improves quality and tolerability, but only controlled and validated processes can lead to gluten levels compatible with a gluten-free diet.

For those who are not celiac but seek products that are more digestible, less stressful for the intestine and based on natural fermentation processes, sourdough currently represents one of the most interesting solutions supported by scientific literature.

The Science Behind Bread and Pizza

Chapter I – Gliadins and Glutenins: the essential building blocks
Chapter II – Fermentation in professional baking and pizzeria production
Chapter III – Gluten degradation during fermentation
Chapter IV – Scientific evidence and application limits

Gluten Degradation During Fermentation (Chapter III)

by luciano

Fermentation, Proteolysis and Potential Modulation of Mucosal Stimuli

1. Technical premise

Physiological evidence shows that some protein peptides resistant to digestion can:

  • modulate paracellular permeability

  • activate innate immunity pathways

  • interact with the intestinal microbial ecosystem

In the context of professional baking, the technological interest is not clinical but biochemical and structural: reducing the fraction of peptides relatively resistant to enzymatic digestion and modifying digestive kinetics through appropriately designed fermentation.

It should be emphasized that the primary function of protein digestion is the hydrolysis of dietary proteins — including gluten — into free amino acids and small peptides (mainly di- and tripeptides), which can cross the intestinal epithelium via specific transport systems and be used as metabolic substrates for the various metabolic functions of the body.

The peptide fraction that is not completely hydrolyzed, consisting of larger peptides, nor absorbed at the level of the small intestine, reaches the colon where it is partially metabolized by the intestinal microbiota through fermentative processes; the unused portion is eliminated with feces.

Enzymatic hydrolysis therefore represents the key step for making proteins nutritionally available and limiting the presence of peptide fractions relatively resistant to digestion.

2. How fermentation can act on resistant peptides

2.1 Acidification and enzymatic activation

Sourdough fermentation leads to:

  • pH reduction (≈ 3.8–4.8 depending on the system)

  • activation/modulation of endogenous flour proteases

  • production of microbial peptidases

Resulting effect:

  • reduction of the average molecular weight of protein fractions

  • increase in the pool of short peptides and free amino acids

  • remodeling of the peptide profile

This does not correspond to “elimination of gluten,” but to modification of the distribution of protein fragments (greater quantity of short peptides).

2.2 Depolymerization of the gluten network

Prolonged fermentation can:

  • reduce the gluten macropolymer

  • modify the secondary structure of proteins

  • make the network less compact and more accessible to digestive enzymes

Potential physiological consequence:

  • improved accessibility to gastric/pancreatic proteolysis

  • reduction of the fraction of persistent long peptides

2.3 Time as a critical variable

The maturation time is determinant:

Short time

Prolonged time

Prevalence of gas development

Greater proteolysis

Network still compact

Greater protein reorganization

Peptide profile little modified

Distribution toward shorter peptides

In professional practice, fermentations of 24–72 h at controlled temperature increase the probability of significant but structurally controlled proteolysis.

3. Baker’s yeast vs sourdough

Baker’s yeast (Saccharomyces cerevisiae)

  • limited proteolytic activity

  • mainly indirect effect (time, hydration, activation of flour enzymes)

  • reduction of resistant peptides mainly dependent on maturation time

Sourdough (LAB + yeasts)

  • direct peptidase activity

  • structuring acidification

  • greater protein remodeling at equal time

4. Interaction with microbiota and intestinal barrier

In light of physiological and experimental evidence, there is in vitro and murine model evidence suggesting a possible systemic impact of gluten on intestinal permeability and inflammatory balance, particularly in subjects with genetic predisposition, immunological vulnerability or pre-existing clinical conditions.

In this context, long and resistant peptides derived from gluten may interact with the intestinal barrier and innate immunity, influencing their functionality. Such interaction may translate into modifications of intestinal permeability, variations in microbiota composition and modulation of immune responses.

Therefore, a criterion of nutritional prudence does not represent excessive caution but rather an act of preventive responsibility.

In healthy individuals there are currently no solid and conclusive clinical data demonstrating a significant systemic impact of gluten on intestinal permeability or inflammatory balance.

The real effect also depends on:

  • the state of the intestinal mucosa

  • the composition of the microbiota

  • the overall composition of the meal

  • stress level and lifestyle

  • exposure to environmental contaminants

* By “possible impact” it is meant that the interaction between gluten and the organism is closely related to the state of the subject and to their overall biological context. Numerous factors — diet, stress, lifestyle habits and environment — may influence the outcome.

** Finally, it must be specified that by “healthy subject” we do not simply mean an individual without clinically manifest diseases, but a person without ongoing pathologies and without a state of chronic low-grade inflammation. This distinction is fundamental, since in clinical practice the term “healthy” is often used in a limited sense, coinciding only with the absence of formal diagnoses.

5. Digestibility as a property of the food matrix

It is essential to reiterate:

Digestibility is not a property of the protein or starch fraction alone, but of the entire food matrix.

Factors influencing the real digestion of the finished product include:

  • fibers (bran, arabinoxylans)

  • lipids

  • final hydration

  • alveolar structure

  • protein–starch interaction

  • baking method

The presence of fibers, for example, modifies the digestive kinetics of starch and proteins much more than a simple variation in protein content would.

6. Practical implications for the professional

If the goal is to obtain a product with:

  • high biochemical maturation

  • more evolved protein profile

  • lower fraction of peptides relatively resistant to digestion

the design levers are:

  1. reduction of yeast dosage

  2. controlled extension of fermentation

  3. use of well-managed sourdough

  4. control of temperature and pH

  5. balance between proteolysis and structural stability

7. Technical conclusion

In traditional baking:

Prolonged fermentation and controlled acidification can remodel the peptide profile of gluten. This remodeling may reduce the fraction of protein fragments relatively resistant to digestion. Physiological evidence shows that such fragments, in experimental models, can modulate barrier function and innate immunity. Direct transfer of these results to healthy humans requires interpretative caution.

Chapter IV – Scientific Evidence and Applicative Limits

1. Scope and operational definitions

In technical language it is essential to separate three concepts that are often confused:

1. Gluten hydrolysis/proteolysis
→ fragmentation of proteins (gliadins and glutenins) into smaller peptides.

  1. Reduction of immunogenic peptides/epitopes for celiac disease→ degradation of specific sequences rich in proline and glutamine (e.g. “Pro-rich” peptides) that resist digestion and activate immune responses in celiac patients.

  1. “Elimination” of gluten→ a much more ambitious objective, achievable only under controlled technological conditions (selected strains, often enzymatic co-adjuvants, long fermentation times), and not equivalent to normal baking with traditional sourdough.

2. Evidence: what studies show

2.1 Fermentation with selected lactic acid bacteria: targeted degradation of immunogenic peptides

Di Cagno et al., 2004 (Applied and Environmental Microbiology) demonstrate that the use of selected lactobacilli with specialized peptidases is able to hydrolyze proline-rich peptides, including peptides with high immunogenicity (the work explicitly discusses the hydrolysis of “Pro-rich” peptides and the application to an experimental baked product).
The study also includes an acute clinical challenge test in subjects with celiac disease within the described experimental protocol. (PubMed)

Key technical points (what is “demonstrated”)

  • The ability to degrade prolamin fractions critically depends on strain selection (it is not an automatic effect of any sourdough). (PubMed)

  • Degradation involves peptides known to resist gastrointestinal digestion thanks to enzymatic systems (peptidases) not typical of baker’s yeast alone. (PubMed)

Immediate applicative limit

The protocol is not “generic sourdough”: it is a biotechnology using selected strains and defined conditions; it is not automatically transferable to any artisanal process. (PubMed)

2.2 “Enhanced” fermentation: selected lactobacilli + fungal proteases (extensive detoxification)

Rizzello et al., 2007 (Applied and Environmental Microbiology) show an even more “engineered” approach: a mixture of selected lactobacilli + fungal proteases during prolonged fermentation.

The study uses several analytical techniques (immunological and instrumental) to estimate residual gluten and the persistence of different protein fractions. (PubMed)

Key technical points

  • Complete hydrolysis of gliadins and other soluble fractions reported in the experimental process; partial persistence of a fraction of glutenins (not all structural fractions are necessarily “eliminated”). (PubMed)

  • Measurement of residual gluten through immunological tests (R5-ELISA) and confirmation through proteomic/spectrometric analyses in the protocol. (PubMed)

  • Biological evaluation of immunoreactivity (tests on immune cell lines) to estimate the “toxicity” of the pepsin-trypsin digest of the fermented product. (PubMed)

Applicative limit

This scenario requires enzymatic co-adjuvants (fungal proteases) and a controlled setup: it is an industrial/biotechnological process, not the equivalent of standard sourdough management in a bakery. (PubMed)

2.3 Selected lactic fermentation on different cereals: role of pH and endogenous enzymes

De Angelis et al., 2006 (Journal of Cereal Science) study the fermentation of rye flours with selected lactic acid bacteria, showing extensive hydrolysis of ethanol-soluble polypeptides and a reduction of immunochemical detectability (R5-Western), also discussing the role of pH in activating hydrolysis through endogenous flour enzymes. (ScienceDirect)

Key technical points

The observed degradation results from a combination of:

  • microbial proteolytic activity (selected strains)

  • pH-dependent hydrolysis (activation of endogenous cereal enzyme systems) (ScienceDirect)

The work supports the “biotechnological” logic of controlled fermentation as a tool to reduce contamination/reactivity risk in specific contexts (in experimental terms). (ScienceDirect)

Applicative limit

Again: selected strains + defined process conditions; this is not an automatic generalization for “any sourdough.” (ScienceDirect)

3. Where baker’s yeast and “traditional” sourdough fit

3.1 Baker’s yeast (Saccharomyces cerevisiae)

Within the framework of the above studies, the effect of baker’s yeast is mainly:

  • fermentative kinetics (CO₂, volumetric development)

  • indirect influence on maturation (time/temperature)

but not a proteolytic activity comparable to that of selected lactic bacteria and/or added proteases.

In other words: with baker’s yeast the “improved digestive management” (when observed) is more related to maturation time and transformations of the starch-protein matrix, not to extensive degradation of immunogenic gluten sequences (in the terms used in the cited studies).

(This is a conclusion derived by comparing the mechanisms reported in studies on selected LAB and proteases.) (PubMed)

3.2 “Non-selected” sourdough (spontaneous sourdough starter)

Spontaneous sourdough can determine:

  • acidification

  • partial proteolysis

  • rheological modifications

However, the literature showing “almost total” degradation or marked reduction of immunogenic epitopes typically uses:

  • selected lactic strains with specific peptidases (PubMed)

and/or

  • fungal proteases in combination (PubMed)

Therefore, at a manualistic level, the correct formulation is:

Fermentation with sourdough can increase gluten proteolysis; extensive degradation of immunogenic sequences requires controlled biotechnological protocols (selected strains and, in some cases, enzymatic co-adjuvants).

4. Applicative limits

Protocols aiming to drastically reduce the immunogenic fraction of gluten do not coincide with the standard production of sourdough bread/pizza. (PubMed)

The result depends on:

  • microbial species/strains used (selection) (PubMed)

  • fermentation time

  • acidity/pH (and relative enzymatic activation) (ScienceDirect)

  • possible use of technological proteases (PubMed)

Even when very extensive degradation is observed, some studies report possible persistence of certain fractions (e.g. part of the glutenins) depending on the protocol. (PubMed)

5. Cited studies

Di Cagno, R. et al. (2004). Sourdough bread made from wheat and nontoxic flours and started with selected lactobacilli is tolerated in celiac sprue patients. Applied and Environmental Microbiology, 70(2), 1088–1096. DOI: 10.1128/AEM.70.2.1088-1096.2004 (PubMed)

Rizzello, C.G. et al. (2007). Highly efficient gluten degradation by lactobacilli and fungal proteases during food processing: new perspectives for celiac disease. Applied and Environmental Microbiology, 73(14), 4499–4507. DOI: 10.1128/AEM.00260-07 (PubMed)

De Angelis, M. et al. (2006). Fermentation by selected sourdough lactic acid bacteria to decrease coeliac intolerance to rye flour. Journal of Cereal Science, 43(3), 301–314. DOI: 10.1016/j.jcs.2005.12.008 (ScienceDirect)

In-depth analysis

Effects of sourdough and/or yeast use in gluten fermentation: scientific evidence

Primary studies (main evidence)

1. Effects of LAB + yeast co-fermentation on gluten degradation

Title: Effects of Co-Fermentation with Lactic Acid Bacteria and Yeast on Gliadin Degradation in Whole-Wheat Sourdough

Summary: The study evaluates how selected strains of Lactic Acid Bacteria (LAB) and baker’s yeast (Saccharomyces cerevisiae) co-ferment gluten in whole-wheat sourdough. The combined fermentation leads to significant degradation of gliadin and glutenin fractions, with reduction of gluten content. Strains such as Lactobacillus brevis and Pediococcus pentosaceus show high proteolytic activity. (MDPI)

2. Reduction of gluten allergenicity in fermented products

Title: From gluten structure to immunogenicity: Investigating the effects of lactic acid bacteria and yeast co-fermentation on wheat allergenicity in steamed buns

Summary: LAB + baker’s yeast co-fermentation induces depolymerization of gluten macromolecules and reduces total immunoreactivity compared with non-fermented controls. Significant decreases in α/γ-gliadins and glutenins associated with celiac disease are observed. (PubMed)

3. Immunogenic peptides and sourdough

Title: A Case Study of the Response of Immunogenic Gluten Peptides to Sourdough Proteolysis

Summary: Fermentation with sourdough modifies gluten structure and the release profile of immunogenic peptides during in vitro digestion, without necessarily eliminating them completely. Comparative study between sourdough bread and rapid-leavened bread. (PubMed)

4. Bacillus spp. isolated from sourdough and gluten hydrolysis

Title: Gluten hydrolyzing activity of Bacillus spp isolated from sourdough

Summary: Bacillus strains isolated from sourdough degrade the immunogenic 33-mer peptide and gliadin sequences, reducing gluten below 110 mg/kg. Potential application in reduced-gluten products. (SpringerLink)

5. Pilot clinical study on fermented products

Title: Gluten-free sourdough wheat baked goods appear safe for young celiac patients: a pilot study

Summary: Fermentation with selected lactobacilli and fungal proteases reduces gluten below 10 ppm. Products tested on children with celiac disease in remission show good clinical tolerability. (PubMed)

6. Recent review on the role of fermentation (2025)

Title: Sourdough Fermentation and Gluten Reduction: A Biotechnological Approach for Gluten-Related Disorders

Summary: LAB fermentation contributes to the reduction of gluten peptides but is not sufficient alone to eliminate all immunogenic sequences. Combined processes with exogenous proteases are more effective. (MDPI)

Previously cited studies, with greater detail

A. Bacillus spp isolated from sourdough
DOI: 10.1186/s12934-020-01388-z

Further detail: The study demonstrates the high proteolytic activity of Bacillus strains against gliadin substrates and the 33-mer peptide. Extensive hydrolysis leads to gluten levels <110 mg/kg in fermented sourdough.

B. Label-free quantitative proteomics and sourdough fermentation
DOI: 10.1016/j.foodchem.2023.137037

Further detail: Proteomic analysis identifies 85 allergenic proteins modulated by fermentation. Some microbial combinations show reduction of gliadins containing immunogenic sequences, suggesting a selective effect of fermentation on the wheat protein fraction.

C. Yeast–bacteria interactions and immunogenicity
DOI: 10.1016/j.ifset.2023.103281

Further detail: Co-cultures of yeasts (Saccharomyces, Torulaspora) with Pediococcus acidilactici show greater gluten depolymerization and reduced immunogenicity compared with single-yeast fermentations.

General conclusions

Sourdough fermentation can partially degrade gluten and reduce specific immunogenic peptides. The reduction does not equal complete elimination: without exogenous proteases, residual gluten often remains. Effectiveness strongly depends on microbial strains and fermentation conditions.

What does all this mean for those seeking gluten-light products?

Products made with sourdough (sourdough fermentation) generally present technological and biochemical characteristics superior to products obtained with rapid leavening, especially regarding tolerability and overall quality.

In particular:

Partial gluten degradation

Prolonged fermentation promotes hydrolysis of some gliadin and glutenin fractions, reducing protein complexity compared with non-fermented doughs.

Modified peptide profile

Even when gluten is not eliminated, its structure changes, with a potential reduction of specific immunogenic peptides.

Perceived improved digestibility

Many non-celiac consumers report better gastrointestinal tolerance compared with industrial baked products produced with rapid fermentation.

Reduction of other critical factors

Sourdough fermentation also contributes to decreasing FODMAPs and some antinutritional compounds.

⚠️ Important note: gluten-light products are not automatically safe for people with celiac disease. Traditional fermentation improves quality and tolerability, but only controlled and validated processes can lead to gluten levels compatible with a gluten-free diet.

For those who are not celiac but seek products that are more digestible, less stressful for the intestine and based on natural fermentation processes, sourdough currently represents one of the most interesting solutions supported by scientific literature.

The Science Behind Bread and Pizza

Chapter I – Gliadins and Glutenins: the essential building blocks
Chapter II – Fermentation in professional baking and pizzeria production

Chapter III – Gluten degradation during fermentation
Chapter IV – Scientific evidence and application limits

The Science Behind Bread and Pizza (Chapter I and II)

by luciano

sangiorgio.l@libero.it

Biochemistry, Rheology and Microbiology of Fermentation and the Starch–Protein Matrix

The present text analyzes the biochemical, rheological and microbiological foundations underlying the production of bread and pizza. The role of gluten proteins (gliadins and glutenins), fermentative systems (baker’s yeast and sourdough), dosage and time variables, and direct and indirect dough-making methods are examined. The approach adopted is technological-functional, with particular attention to the structural, aromatic, digestive and shelf-life implications of the finished product.

Chapter I – Protein Architecture of Dough: Gliadins, Glutenins and the Gluten Network

When we mix flour and water, we are not simply combining ingredients: we are activating a complex protein system that determines structure, consistency and the final result.
At the base of everything is gluten, a three-dimensional network created by the interaction between two families of wheat proteins: gliadins and glutenins. Understanding their balance means understanding why a pizza dough stretches easily while bread dough must sustain a tall, aerated structure.

1️⃣ The Gluten Network: A Dynamic Balance

Gluten does not exist “already formed” in flour. It is created when:

Glutenin + Gliadin + Water + Mixing = Gluten network

Water hydrates the proteins, the mechanical energy of mixing makes them interact, and an elastic network capable of trapping fermentation gases is formed. But the two proteins perform different and complementary roles.

2️⃣ The Role of Glutenins: Strength and Elasticity

Glutenins – Structural Effects

Glutenins provide:

Elasticity (ability to return to the original shape)
Tenacity (resistance to deformation)
Structure

A dough rich in glutenins:

Is more resistant
Retains gases better
Develops vertical volume

If excessive:

Too tenacious
Difficult to stretch
“Spring-back” effect

3️⃣ The Role of Gliadins: Extensibility and Viscosity

Gliadins are responsible for:

Extensibility (ability to stretch without tearing)
Malleability
Viscosity

Thanks to gliadins, the dough:

Stretches easily
Does not tear during handling
Maintains good workability

If they dominate excessively, however, the dough:

Becomes soft
“Spreads”
Struggles to maintain shape

4️⃣ Pizza: Extensibility Is Required

In the case of pizza, the goal is to obtain a thin disk that:

Stretches easily
Does not tear
Does not retract during shaping

Extensibility is therefore fundamental. A dough too rich in glutenins would be “rubbery” and difficult to open.

For this reason, pizza flours (often soft wheat) are designed to have:

A good balance between strength and extensibility
A P/L ratio (tenacity/extensibility) balanced or slightly shifted toward extensibility

If the dough is too tenacious, it is possible to intervene with:

Longer maturation (longer resting time)
Increased hydration
Choosing a flour with a lower P/L ratio

In summary: more extensibility = easy stretching and good alveolation.

5️⃣ Bread: Structural Strength Is Required

Why does bread need more glutenins?

Bread has a different goal: to develop vertically and sustain an internal structure rich in alveoli.

Here the following come into play:

Elasticity
Structural strength
Capacity to retain fermentation gases

Bread dough therefore requires a stronger gluten network, with a higher glutenin component.

If gliadins dominate too much:

The dough becomes weak
It spreads instead of rising
The bread results low and poorly structured

In summary: more glutenins = more strength and vertical development.

6️⃣ Balance Is the Key

The fundamental point is not “which protein is better”, but their ratio.

Too much glutenin → tenacious dough, hard to stretch
Too much gliadin → soft and unstable dough
Correct balance → elastic and extensible structure

The difference between pizza and bread lies precisely in this balance:

Product

Dominant characteristic

Protein ratio

Pizza

Extensibility

Good presence of gliadins

Bread

Strength and elasticity

Greater glutenin component

7️⃣ Conclusion

Pizza → more extensibility (gliadins)
Bread → more strength and elasticity (glutenins)

The quality of a dough does not depend only on the quantity of proteins, but on their interaction, processing, hydration and maturation time. Every time we stretch a pizza or shape a loaf of bread, we are working with a delicate molecular balance: a true protein architecture that transforms flour and water into a living, elastic and extensible structure.

Chapter II – Fermentation in Professional Baking and Pizzeria Production

Role of baker’s yeast and sourdough, quantities, time and dough-making methods

1. Introduction

Fermentation represents the biological and technological core of professional baking and pizza production. It is not limited to the production of gas for dough volume increase, but profoundly determines:

Mechanical structure
Extensibility
Alveolation
Aromatic profile
Digestibility
Shelf life

The professional does not simply manage a “leavening”, but a complex biochemical process in which interact:

Microorganisms
Endogenous flour enzymes
Gluten proteins
Starches
Time
Temperature

This chapter systematically analyzes the role of baker’s yeast and sourdough, the influence of dosage and fermentation time, and the impact of processing methods (direct dough and indirect dough with biga) on the finished product.

2. The Role of Baker’s Yeast

2.1 Microbiological Nature

Baker’s yeast consists predominantly of Saccharomyces cerevisiae, a unicellular microorganism capable of metabolizing simple sugars present in dough.

Alcoholic fermentation produces:

Carbon dioxide (CO₂)
Ethanol
Secondary metabolites (esters, higher alcohols, aldehydes)

CO₂ is retained by the gluten network and generates the increase in volume.

2.2 Technological Effects

Baker’s yeast:

Provides gas for structural development
Indirectly stimulates enzymatic activity
Influences fermentation rate
Determines part of the aromatic profile

It does not significantly modify dough pH (limited acidity), therefore the effect on the protein structure is mainly mechanical and fermentative, not acidifying.

3. The Role of Sourdough

3.1 Microbiological Nature

Sourdough is an ecosystem composed of:

Wild yeasts
Lactic acid bacteria (homo- and hetero-fermentative)

These microorganisms produce:

CO₂
Lactic acid
Acetic acid
Proteolytic enzymes
Complex aromatic compounds

3.2 Technological Effects

The combined activity of yeasts and bacteria

The combined activity of yeasts and bacteria determines

The controlled acidity of yeasts and bacteria directly influences

Progressive acidification (pH reduction)
Elasticity
Modification of gluten structure
Extensibility
Activation of proteases
Shelf life
Improved microbiological stability
Aromatic depth

4. Quantity and Time: General Principles

4.1 Relationship Between Dosage and Speed

The quantity of fermenting agent regulates:

CO₂ production speed
Metabolic intensity
Process duration

Fundamental principle:

More yeast → rapid fermentation
Less yeast → slow fermentation

However, speed does not coincide with maturation.

4.2 Time as a Key Variable

Time allows:

Enzymatic degradation of starches (amylases)
Partial protein hydrolysis
Reorganization of the gluten network
Formation of aromatic metabolites

A short fermentation may produce volume, but not necessarily structural and biochemical maturation.

5. Effects on Digestibility

5.1 Technical Definition

Digestibility refers to:

Reduction of intestinal fermentable load
Partial pre-digestion of starches and proteins
Better structural organization of the crumb

It does not imply absence of gluten, but a more advanced biochemical transformation.

5.2 Baker’s Yeast

Baker’s Yeast Dosage

High dosage + short time
Low dosage + long time

Limited maturation
Greater maturation

Lower enzymatic activity
Better enzymatic degradation

Higher presence of residual sugars
Biochemically evolved dough

Possible sensation of heaviness
Greater sensation of lightness

5.3 Sourdough

Fermentation with sourdough determines:

Progressive reduction of pH (controlled acidification)
Increase of proteolytic activity (endogenous enzymes + microbial activity)
Partial hydrolysis of gluten proteins
Greater degradation of fermentable sugars
Modification of the rheological properties of the gluten network

Measurable technological and physiological effects

Prolonged sourdough fermentations involve:

Reduction of residual fermentable carbohydrates
Partial protein pre-digestion
Better structural organization of the crumb
Slower glycemic response compared to short fermentations
Greater microbiological stability of the product

The combination of these factors may determine:

Reduction of intestinal fermentative load
Lower intestinal gas production compared to rapidly fermented doughs

Individual physiological response may vary depending on personal conditions, but the biochemical mechanisms described above are objectively measurable.

6. Effects on Pizza and Bread

6.1 Pizza

Structural objectives

High extensibility
Absence of “spring-back” effect
Aerated cornicione
Melt-in-the-mouth texture

Typical strategy

Very low yeast dosage
Long maturation (24–72 hours)
Temperature control

Results

Greater extensibility
More complex aroma
Lower sensation of bloating

6.2 Bread

Structural objectives

Vertical development
Crumb stability
Shelf life

Objectives with baker’s yeast

Regular structure
Delicate aroma

Objectives with sourdough

Irregular alveolation
Thick crust
Deep aroma
Longer shelf life

7. Dough-Making Methods

7.1 Direct Dough

7.1.1 Definition

All ingredients are mixed in a single phase.

7.1.2 Fermentation Dynamics

Immediate complete hydration
Single fermentation
Structure progressively built

7.1.3 Effects on the Product

Effects on the product

Texture
Homogeneous crumb
Regular alveolation

Aroma
Linear profile
Lower complexity

Digestibility
Good if accompanied by long fermentation times
Lower if fermentation is short

Shelf life
Faster staling
Lower protective acidification

7.2 Indirect Dough with Biga

7.2.1 Definition

Solid preferment (45–50% hydration) with:

Flour
Water
Small quantity of yeast

Fermentation 16–24 hours before the final dough.

7.2.2 Biochemical Dynamics

During biga maturation:

Early enzymatic activation
Production of light organic acids
Pre-maturation of gluten
Development of aromatic precursors

7.2.3 Effects on the Product

Effects on the product

Texture
Large and irregular alveolation
Greater lightness
Crispier crust

Aroma
Greater complexity
Light lactic aromas
Intensification of toasted notes

Digestibility
Dough already partially matured
Lower residual fermentable load

Shelf life
Better moisture retention
Slower staling
Greater aromatic persistence

8. Systemic Comparison

Variable

Direct

Biga

Structure

Regular

Airy and light

Aroma

Linear

Complex

Digestibility

Depends on time

Generally higher

Shelf life

Medium

Higher

Management complexity

Low

Medium/High

9. Integrated Design Principle

In professional contexts, dough design simultaneously considers:

Type of fermenting agent
Dosage
Time
Temperature
Method (direct or indirect)

There is no universally superior solution, but rather a balance consistent with:

Product identity
Sensory objectives
Desired structure
Production organization

10. Conclusion

Fermentation is not an accessory step, but a process of structural and biochemical transformation. The quantity of yeast, the choice between baker’s yeast and sourdough, the maturation time and the adopted method (direct or biga) constitute tools of applied food engineering. The professional does not simply “let a dough rise”: they design the behavior of matter over time in order to obtain a structural, aromatic and functional result consistent with the identity of the final product.

Essential Bibliography

Gluten, Protein Structure and Rheology

  1. Wieser, H. (2007).
    Chemistry of gluten proteins.
    Food Microbiology, 24(2), 115–119.
    DOI: 10.1016/j.fm.2006.07.004

  2. Shewry, P. R., & Tatham, A. S. (1997).
    Disulphide bonds in wheat gluten proteins.
    Journal of Cereal Science, 25(3), 207–227.
    DOI: 10.1006/jcrs.1996.0100

  3. Belton, P. S. (1999).
    On the elasticity of wheat gluten.
    Journal of Cereal Science, 29(2), 103–107.
    DOI: 10.1006/jcrs.1998.0233

  4. Dobraszczyk, B. J., & Morgenstern, M. P. (2003).
    Rheology and the breadmaking process.
    Journal of Cereal Science, 38(3), 229–245.
    DOI: 10.1016/S0733-5210(03)00059-6

Baker’s Yeast and Alcoholic Fermentation

  1. Fleet, G. H. (2007).
    Yeasts in foods and beverages: impact on product quality and safety.
    Food Microbiology, 24(2), 103–112.
    DOI: 10.1016/j.fm.2006.07.002

  2. Walker, G. M. (1998).
    Yeast Physiology and Biotechnology.
    John Wiley & Sons.
    ISBN: 978-0471964467

Sourdough and Sourdough Microbiology

  1. De Vuyst, L., & Neysens, P. (2005).
    The sourdough microflora: biodiversity and metabolic interactions.
    Trends in Food Science & Technology, 16(1–3), 43–56.
    DOI: 10.1016/j.tifs.2004.02.012

  2. Gobbetti, M., De Angelis, M., Di Cagno, R., Calasso, M., & Archetti, G. (2019).
    Novel insights on the functional/nutritional features of sourdough fermentation.
    International Journal of Food Microbiology, 302, 103–113.
    DOI: 10.1016/j.ijfoodmicro.2018.05.018

  3. Poutanen, K., Flander, L., & Katina, K. (2009).
    Sourdough and cereal fermentation in a nutritional perspective.
    Food Microbiology, 26(7), 693–699.
    DOI: 10.1016/j.fm.2009.07.011

  4. Hammes, W. P., & Gänzle, M. G. (1998).
    Sourdough breads and related products.
    Food Microbiology, 15(5), 487–495.
    DOI: 10.1006/fmic.1998.0191

Focus on Digestibility

1️⃣ Arendt et al., 2007

Impact of sourdough on the texture of bread
Food Microbiology, 24(2), 165–174.

Study Objective

Analyze the effect of sourdough fermentation on:

Crumb structure
Texture
Starch retrogradation
Shelf life

Key Points Relevant to Digestibility

  1. Controlled acidification

Reduction of pH
Influence on starch gelatinization and retrogradation

  1. Modification of starch structure

Acidity slows retrogradation
Better water retention
Greater crumb stability

  1. Gluten–starch interaction

Acid fermentation modifies the protein matrix
Better starch distribution within the gluten network

Implications for Digestibility

Digestibility is influenced through:

Greater enzymatic accessibility to starch
Less compact and less collapsed structure
More modulated carbohydrate release

In technical terms: acid fermentation modifies the microstructure of the starch–protein matrix, influencing digestive kinetics.

2️⃣ Liljeberg & Björck, 1998

Delayed gastric emptying rate may explain improved glycaemia…
European Journal of Clinical Nutrition, 52(5), 368–371.

Study Objective

Evaluate the effect of food acidity on post-prandial glycemic response.

Key Points

  1. Reduction of meal pH

Slows gastric emptying

  1. More gradual glycemic response

Lower glycemic peak
Better control of glucose absorption

  1. Physiological mechanism

A more acidic environment modifies digestion and absorption rate.

Implications for Bread and Pizza

In sourdough bread:

Lower pH
Presence of organic acids

may contribute to:

Slowing digestive kinetics
Reducing the rate of glucose release

Digestibility does not mean “fewer calories”, but a more modulated metabolic release.

3️⃣ Katina et al., 2006

Effects of sourdough and enzymes on staling of high-fibre wheat bread
LWT – Food Science and Technology, 39(5), 479–491.

Study Objective

Analyze the effect of:

Sourdough
Enzymatic activity
Fiber structure

on:

Staling
Retrogradation
Texture

Key Points

  1. Prolonged enzymatic activity

Greater degradation of starches
Partial hydrolysis of polysaccharide structures

  1. Slower retrogradation

Better crumb stability over time

  1. Enzyme–structure interaction

Greater modification of the structural matrix

Implications for Digestibility

Partially modified starch → different digestive enzymatic response
Greater enzymatic availability
Reduction of residual fermentable substrates

Long fermentation alters carbohydrate structure before baking.

What Is Scientifically Meant by “Digestibility” in Long-Fermented Bread and Pizza

Based on the three studies, it can be defined as:

1️⃣ Structural modification of the matrix

Gluten–starch reorganization
Greater accessibility to digestive enzymes

2️⃣ Reduction of residual fermentable load

Lower presence of rapidly fermentable sugars

3️⃣ Modulation of glycemic response

Slower glucose release
Buffering effect of acidity

4️⃣ Influence on gastric emptying

Lower pH → slower emptying
More gradual absorption

Final Technical Synthesis

Digestibility in long-fermented bread and pizza is obtained through:

Time (enzymatic maturation)
Acidification (sourdough)
Structural modification of starch and proteins
Reduction of residual fermentable load
Modulation of glycemic response

It is a structural biochemical effect, not a merely “perceived” property.

Enzymatic and microbial transformations do not concern only starch and aromas: under specific conditions, they also involve the protein fraction of gluten, reshaping its peptide profile. This topic is addressed in Chapter III.

The Science Behind Bread and Pizza

Chapter I – Protein Architecture of Dough: Gliadins, Glutenins and the Gluten Network
Chapter II – Fermentation in professional baking and pizzeria production

Chapter III – Gluten degradation during fermentation
Chapter IV – Scientific evidence and application limits

Experimental application of the advanced method for the production of bread doughs with flours with limited gluten development capacity: analysis of the results. (Analysis carried out by ChatGPT)

by luciano

sangiorgio.l@libero.it

Related article: Advanced method for making bread dough with flours with limited gluten development capacity

(Dynamics of the protein network in Triticum monococcum bread subjected to prolonged cold maturation and controlled thermal reactivation. Physical-dynamic model, qualitative rheological interpretation, and experimental validation through photographic documentation and evaluation of the finished product).

Technical-scientific synthesis. The present study analyzes the structural and rheological behavior of an einkorn dough (Triticum monococcum L.) subjected to a process structured into biga, dispersion in the liquid phase, prolonged cold maturation, thermal reactivation on a warm surface, intermediate manipulations, final proofing, and baking. The objective is to verify whether the protein network of einkorn follows a linear development dynamic or a non-linear dynamic, characterized by a phase of transient instability followed by possible reorganization. The experiment compared two dough series identical in their general scheme but differing in maturation duration and, above all, in temperature control during the post-retard phase. Series I, conducted with more coherent thermal control, showed recovery of surface continuity, satisfactory vertical development, guided opening of the score, and a structurally functional crumb. Series II, subjected to longer maturation and compromised thermal control, showed a more fragile surface, greater morphological irregularity, less controlled expansion, and a more heterogeneous crumb, though still fully functional from a food standpoint. Based on the images and process data, a six-stage model is proposed: 1)initial aggregation of the preferment; 2)controlled dispersion; 3)cold maturation with biochemical relaxation; 4)critical post-retard instability window; 5)protein reorganization assisted by rest and manipulation; 6)stabilization of the functional network. The qualitative rheological interpretation suggests that the decisive parameter in einkorn is not merely the intrinsic strength of the flour, but the synchronism between reorganization of the protein matrix and gaseous development [1][2]. The theoretical framework is consistent with the literature on wheat gluten, which assigns a central role in dough viscoelasticity to glutenins, gliadins, and glutenin macropolymer, and with the literature on einkorn, which generally describes softer, less elastic, and more extensible doughs than common wheat, albeit with strong dependence on genotype and process [1][3][4]

1. Introduction

Einkorn is one of the oldest domesticated wheat species and is today being rediscovered for its nutritional, agronomic, and sensory interest. However, its breadmaking remains problematic, because its high protein content does not automatically translate into a strong gluten network in the technological sense typical of modern bread wheat. The literature indicates, in fact, that the breadmaking quality of wheat depends not only on total protein quantity, but also on protein composition, the gliadin/glutenin ratio, the presence and functionality of glutenin subunits, and the ability to form high-molecular-weight aggregates capable of conferring elasticity, cohesion, viscosity, and gas retention. Wieser recalls that gliadins are predominantly monomeric and associated mainly with viscosity and extensibility, whereas glutenins are polymeric, aggregated through interchain disulfide bonds, and constitute the component most directly associated with elasticity and the structure of the dough network [1].

In the case of einkorn, the situation is more delicate. Recent literature reports that, despite its high protein content, einkorn flours tend on average to produce softer, less elastic, and more extensible doughs than modern wheats, due to a generally lower gluten quality and a different balance between protein fractions. Brandolini and co-workers observe that einkorn “generally has poor breadmaking value,” although some elite lines show very interesting technological performance; Mefleh and co-workers, in a holistic approach on Italian genotypes, show marked differences in breadmaking behavior between ancient and improved varieties [3][4]. The overall picture suggests that einkorn should not be judged as a species intrinsically incapable of making bread, but as a system highly sensitive to genotype and process.

The present study is situated in this area of interest. It does not aim to generically demonstrate whether einkorn “does” or “does not” make bread, but rather to describe the structural dynamics of its dough during a complex protocol of cold maturation and thermal reactivation. The starting hypothesis is that the protein network of einkorn does not evolve in a linear and monotonic way, but rather passes through a phase of transient instability in which the apparent rupture of the surface may precede a useful reorganization of the matrix. The literature on glutenin macropolymer offers theoretical support for this possibility: Feng and co-workers show that mixing reduces the apparent content of GMP, whereas resting can restore part of it; at the same time, excessive resting can worsen the structural framework again [2].

This scientific framework makes particularly significant the empirical observation that guides the entire work: bread must be judged not only as a form to admire, but as a food material to be eaten. From a technological point of view, the distinction is crucial, because a bread with irregular morphology may still possess a continuous, non-sticky, sufficiently elastic, and sensorily valid crumb. This dual perspective, morphological and functional, guides the entire following analysis.

2. Objectives of the study

The main objective is to verify whether, in einkorn, the protein network during a breadmaking process with cold maturation follows a stage-based dynamic, in which a phase of transient fragility may be followed by a phase of structural reorganization.
The secondary objective is to identify the role of thermal control in the post-retard phase as a possible bifurcation variable between two outcomes: 1)a relatively stable functional network; 2)a still edible but more irregular network.

The third objective is to propose a qualitative rheological interpretation of the images and the behavior of the finished bread, without resorting to direct instrumental tests, while remaining consistent with the literature on the relationships between GMP, viscoelasticity, gas retention, and bread quality [1][2].

3. Materials, experimental setup, and data sources

The study is based on experimental documentation consisting of a written protocol, temperature and pH measurements, pre- and post-baking weights, and a photographic sequence of the main processing stages and the final product.
The raw material is stone-ground whole einkorn flour.

The process includes: 1)initial biga; 2)dissolution/dispersion; 3)cold maturation; 4)reactivation on a warm surface; 5)intermediate manipulations; 6)division; 7)shaping; 8)proofing in a basket; 9)scoring; 10)baking.

Two series were compared.

Series I: 24-hour maturation, ambient temperature approximately 22 °C, surface temperature on removal approximately 6 °C, more coherent management of the warm surface.
Series II: 28-hour maturation, ambient temperature approximately 23 °C, surface temperature on removal approximately 7.8 °C, lower initial pH, and compromised thermal control of the warm surface.
The temperature, pH, time, weight, surface description, and crust and crumb evaluations are those experimentally recorded in the protocol; the scientific interpretations of their meaning are related to the literature [2][3][4]. The images were read as morphological evidence of the mechanical and structural behavior of the dough. This is therefore not a study with standardized instrumental tests of alveography, farinography, dynamic oscillation, or texture profile analysis, but rather a high-resolution observational qualitative technical-scientific analysis.

4. Theoretical framework of reference

The breadmaking quality of wheat derives from a complex protein system in which the gliadin fraction and the glutenin fraction play complementary roles.
1)Gliadins, monomeric, contribute mainly to viscosity and extensibility.
2)Glutenins, aggregated through interchain disulfide bonds, constitute the high-molecular-weight polymers most involved in the elastic response of dough.
Glutenin macropolymer is considered a key component of this system, and several sources link it directly to viscoelastic properties and bread volume. Feng and co-workers report that GMP plays a prominent role in dough properties and breadmaking quality, that mixing reduces its apparent content through depolymerization, and that resting may favor a partial recovery through reformation of disulfide bonds and growth in the fraction of large particles [1][2].
In einkorn this balance proves on average less favorable to mechanical resistance. Brandolini and co-workers describe protein-rich einkorn flours with breadmaking value generally lower than that of the control soft wheat; Mefleh and co-workers highlight that the dough and bread performance of ancient and improved wheats depends on a set of compositional, rheological, and fermentative parameters [3][4]. The conclusion that emerges is therefore not a technological condemnation of einkorn, but the recognition of a strong dependence on genotype and process. This approach fits the present experiment well: two series very close in formulation, but diverging in structural outcome especially in relation to time, pH, and temperature of the post-retard phase.

The distinction between the two series is essential to correctly interpret the results.

Main process parameters distinguishing Series I and Series II during post-retard dough handling.

Parameter

Series I

Series II

Impact

Retard maturation

24 h

28 h

Greater enzymatic degradation in Series II

Temperature on removal from retard

6 °C

7.8 °C

More active dough network in Series II

Initial pH

5.2

4.9

Higher acidity in Series II

Warm surface control

Stable

Unstable

Less controlled dough development in Series II

Ambient temperature

22 °C

23 °C

Faster fermentation in Series II

Series II therefore started from a more degraded structural condition, characterized by higher acidity, increased enzymatic activity, and reduced thermal stability.

Series II therefore starts from a more degraded structural condition, characterized by greater acidity, greater enzymatic activity, and greater thermal instability. This explains the presence of a more degraded surface, widespread ruptures, and more irregular alveolation. The most important experimental finding is that the difference between the two series does not seem to depend primarily on the dough, formula, or fermentative agent, but on the thermal management of the post-retard network reactivation phase.

6. Exerimental results: phase-by-phase readin

6.1.Mature pre-dough

The In the initial images the pre-dough appears compact, fragmented, with sharp fractures and a load-bearing mass (Photo 1; Photo 2). This morphology is compatible with a mature pre-dough that has not collapsed. It is not yet a final bread network, but a pre-organized matrix. From the material point of view, the initial state is neither liquid nor completely plastic: it is a cohesive, locally rigid system, rich in structural potential. The visual characteristics are those of a correctly matured pre-dough (biga): hydration distributed, proteolysis present but not excessive, structure still load-bearing. This is important because the quality of the final network originates here.

6.2. Dissolution or dispersion phase

The dissolution phase shows a viscous, aerated system, with microbubbles and fluid continuity (Photo 3). In structural terms this step is equivalent to a controlled dispersion of the original matrix. It is here that the system loses the macroscopic continuity of the biga but gains mobility. The literature on GMP interprets mixing as a phase capable of partially depolymerizing glutenin aggregates and reducing the average particle size, with an increase in the proportion of smaller and more extractable forms [2]. Visually, medium viscosity, the presence of microbubbles, and a fluid but non-liquid structure are observed. This phase produces disaggregation of the gluten network, distribution of the proteins, and dispersion of GMP. This is exactly the principle of the proposed model.

6.3. Dough after cold maturation

After 24 hours at about 5 °C, in the images of the first series the surface appears relatively uniform and the bottom shows structural continuity (Photo 5; Photo 6). This phase does not suggest collapse, but relaxation. From a biochemical point of view it is reasonable to infer that hydration, acidification, and slow protein modifications continued in the retard. Cold slows but does not cancel such processes. In this phase, according to the model that will emerge later, the dough is not yet “ready” in a mechanical sense: it is a relaxed, biochemically modified system awaiting reactivation. Visual observations show a smooth surface, uniform structure, and signs of relaxation; the bottom presents slight adhesiveness but continuous structure. The most coherent interpretation is that of controlled enzymatic maturation, with active amylases, moderate proteases, and a weakened but not destroyed network.

6.4. Critical window after removal from retard

The images of Series I after about two hours on a warm surface show multiple surface ruptures, discontinuities, and loss of homogeneity of the skin (Photo 7, Series I). This is the most important phase of the entire work, because it suggests that einkorn enters a zone of temporary instability. The surface seems to worsen before improving. In a linear model this would be a sign of failure; in the protocol examined here, however, this fragility appears as a transient stage.
Here the most interesting datum is precisely the behavior of the gluten network. Photo 7 represents the phase of network rupture. The observed sequence is the following: 1)removal from retard with a fragile network; 2)manipulation with partial rupture; 3)warm rest with subsequent reaggregation; 4)more elastic structure. You have therefore demonstrated experimentally that the network can reorganize after rupture.

Series II shows in this same phase a more advanced and more vulnerable picture: greater surface porosity, more marked yielding lines, a more open texture already present, and faster fermentative activation (Photo 2, Series II). The whole is consistent with the process data: 1)4 more hours of maturation; 2)lower pH; 3)higher initial dough temperature; 4)less effective control of the warm surface. The literature does not provide a direct photographic relationship, but it makes the general mechanism plausible: in an einkorn dough, already more extensible and less elastic on average, an excessively rapid reactivation can unbalance the relationship between protein reorganization and gas development [3][4].

It is important to note that surface ruptures do not necessarily constitute an absolute defect. In einkorn they may indicate a more rigid surface network in the presence of active internal fermentation. If the internal network holds, the bread can still expand, as happens in this case.

6.5. Manipulation and reorganization

In Series I, after 2 hours and 30 minutes and manipulations, the surface becomes more continuous, smoother, more legible as a structured skin (Photo 9, Series I). The images indicate that the dough recovers cohesion instead of worsening further. The subsequent division also confirms greater stability of the mass (Photo 10, Series I).

Here lies the strongest theoretical point of the entire experiment. Photo 9 documents the reorganization of GMP. The protein system realigns the chains, reforms disulfide bonds, and builds a new, more elastic network. This is exactly the behavior predicted by the model of dispersion and reaggregation of the gluten network.

In Series II, after manipulation, only a partial improvement of the surface is observed, which nevertheless remains more fragile and porous than in Series I (Photo 3, Series II). This suggests that manipulation had a corrective effect, but not a fully refounding one. The connection with Feng et al. is particularly useful here: the resting following mixing can restore part of GMP, but the response depends strongly on the duration of the phase and on the conditions of the system [2].
When fermentation is already too advanced, manipulation does not completely regenerate the network but also tends to break already developed gas chambers. This leads to more irregular alveoli and a less stable structure.

6.6. Division, shaping, and final proofing

In Series I the divided and shaped doughs maintain shape, volume, and a fairly homogeneous surface (Photo 10, Series I). In the basket and at the end of proofing they show growth, the presence of small discontinuities, but good geometric legibility (Photo 13; Photo 14, Series I). The mass holds well, does not collapse, and maintains structure. This is a clear sign that the network has recovered elasticity.

The images of the proofing baskets show an increase in volume, surface microfractures, and still stable structure. The network appears extensible but not excessively weak. In einkorn this is a very good result.

In Series II, by contrast, already at the beginning of proofing in the basket, widespread points of weakness are observed (Photo 5, Series II); before baking the surface appears further compromised and much closer to the fracture threshold (Photo 6; Photo 7, Series II). This visual datum agrees with the hypothesis that the control of temperatures and times during proofing was compromised. In essence, Series II reaches baking with a surface already close to its own fracture threshold. The surface is already very tense, with small widespread rupture points and less elastic structure: a sign that the network is at the limit of its extension capacity.

6.7. Baking, crust, and final shape

In Series I the bread shows good vertical development, wide but legible opening, and orderly growth (Photo 16; Photo 17; Photo 18; Photo 19, Series I). The fracture follows the score coherently and no marked lateral spread is observed. Here we see good expansion, controlled opening, and uniform crust. The fissure follows the score: this indicates functional oven spring.
Vertical development is one of the most important signals of the entire test. It means that the network was extensible but also resistant and that fermentation did not destroy the structure. If the network had been too degraded, broad bread, flat bread, or collapse would have been observed. Instead, the experimental data show final heights between 7 and 7.5 cm for loaves of about 780 g: a very good result.

In Series II the bread instead shows more violent openings and more widespread fractures, with less control of expansion (Photo 8, Series II). However, and this is fundamental to emphasize, neither series shows the catastrophic collapse typical of a completely failed network. No flattened bread, fully slumped bread, or bread lacking internal crumb is observed. The difference is therefore one of control and uniformity, not of the existence or non-existence of structure.
In Series II no true progressive oven spring is observed, but rather a more explosive structural rupture. This is consistent with a situation in which the internal gas exceeds the resistance of the network.

6.8. Crumb

The crumb of Series I appears fine-medium, sufficiently distributed, with some irregularity also attributable to incorporated air, but without large anomalous cavities or massively compact zones (Photo 20; Photo 21, Series I). The bottom of the bread also confirms a well-supported structure (Photo 22, Series I). The bottom shows complete baking, the absence of compressed zones, and the absence of collapse, indicating a good balance between internal structure and hydration/baking ratio.
The description of the crumb is very clear: fine-medium alveoli, fairly homogeneous distribution, elastic crumb, slightly moist, absence of sticky crumb. This means that starch degradation was controlled, the malt is not excessive, and gelatinization occurred correctly. This is a very important finding for einkorn.

Series II presents a more irregular but still valid crumb (Photo 9, Series II). Medium and small alveoli remain predominant, the walls are in most cases continuous, and no sticky crumb or widespread collapse is observed. This implies that the network, although more disordered, retained gas sufficiently to generate a functional food structure. This point is extremely important: Series II is not aesthetically optimal, but it is not technologically null. It is an edible bread, endowed with crumb, structure, and sufficient integrity for food use.

The crumb of Series II deserves a specific evaluation. Despite the degradation of the network, the bread remains functional and edible. The alveolar structure shows predominantly medium-small alveoli, some isolated larger alveoli, non-uniform distribution, but absence of large empty cavities. This indicates that gas retention occurred, that the gluten network did not collapse, and that fermentation produced gas while still maintaining a certain structural cohesion. The crumb is granular but continuous; it does not appear gummy or sticky. The alveolar walls, although thin, are continuous and show good moisture distribution. This suggests an elastic and non-crumbly crumb.
The irregularity of the structure is the sign of the fermentative problem: less uniform alveoli, some isolated larger ones, less ordered structure compared with Series I. The probable causes are: 1)faster fermentation; 2)manipulation on already gassed dough; 3)partial rupture of gas chambers with random recombination of the bubbles.

From a technological point of view, Series II is therefore a valid but less controlled bread. In synthetic terms: good structure, good gas retention, elastic texture, medium homogeneity, irregular aesthetics. Series II demonstrates that even with excessively high temperature, accelerated fermentation, and a more fragile network, einkorn can still produce a stable and edible crumb. The protein system does not collapse completely, but loses part of its structural control.

7. Final quantitative parameters

The The quantitative parameters confirm the morphological and structural interpretation.
For Series I, the pre-baking weight of the loaves was approximately 780 g. The post-baking cold weight was around 652 g. Weight loss is therefore approximately 16-17%, a value fully consistent with a well-baked bread, with balanced internal moisture and a correctly formed crust. For rustic breads and ancient flours, a range of 15-18% can be considered very good.
The recorded final heights, between 7 and 7.5 cm, are also very positive for loaves of this mass.
The recorded final internal temperature was 93.6°C. This value falls within the ideal range of 92-96°C for breads with medium-high hydration and ancient flours, confirming the correctness of the baking.

8. Proposed physical-dynamic model

In light of the images, the data, and the theoretical framework, the behavior of einkorn in the protocol under examination is better described by a six-stage model than by a simple scheme of progressive development.

8.1. State A: initial aggregation of the pre-dought

The mature biga represents a fragmented but load-bearing protein pre-matrix (Photo 1; Photo 2). Water has not yet been redistributed in the way typical of the final dough, but a useful organization already exists. This state is locally stable, even if not yet suitable for final gas retention.

8.2. State B: controlled dispersion

The liquid or dissolution phase reduces macroscopic rigidity and increases the mobility of the components (Photo 3). From the GMP point of view, it is compatible with a partial depolymerization induced by shear and hydration. This does not constitute a definitive loss of function, but a temporary lowering of continuity [2].

8.3. State C: cold maturation and biochemical relaxation

During the stay in the retard, the matrix relaxes, acidifies, and hydrates more finely (Photo 5; Photo 6). The system does not strengthen mechanically in a simple sense, but predisposes itself to a new organization. The low temperature slows the processes but does not cancel them.

8.4. State D: critical post-retard instability window

This phase coincides with the emergence of surface ruptures and discontinuities (Photo 7, Series I; Photo 2, Series II). It is the point at which the system passes from a cold, relaxed, and biochemically modified network to a network once again mobile, fermentatively active, and mechanically stressed. In this window einkorn manifests its typical fragility: if reactivation is well synchronized, the instability is transient; if it is too rapid or too prolonged, the instability is amplified [3][4].

8.5. State E: assisted protein reorganization

Manipulation and rest, within the correct window, allow a recomposition of surface continuity in Series I (Photo 9; Photo 10, Series I), whereas in Series II they produce only a partial reorganization (Photo 3, Series II). The term “reaggregation of GMP” must be used as a structural inference consistent with the literature on resting, not as a direct measurement. The images of Series I and the work of Feng et al. make this interpretation highly plausible [2].

8.6. State F: stabilization of the functional network

The final result bifurcates into two outcomes:

1)relatively stable functional network, with orderly growth and a sufficiently homogeneous crumb (Photo 17; Photo 18; Photo 19; Photo 20; Photo 21, Series I);

2)still functional but less uniform network, with less controlled growth and a heterogeneous crumb (Photo 8; Photo 9, Series II).

This second outcome is precisely that of Series II. The crucial point is that the model does not distinguish only between “success” and “failure,” but between optimal network, functional network, and collapse. In the present experiment the first two levels were observed, not the third.
In synthetic form, the behavior of the dough follows the cycle: dispersion → rupture →

When the temperature is correct: network → dispersion → reaggregation → stable structure.
When the temperature is too high: network → dispersion → degradation → rupture.

9. Qualitative rheological interpretation.

9.1. Methodological premise

Here the term “rheology” is used in a qualitative sense. We do not have instrumental curves of G’, G’’, tan delta, farinograms, or alveograms relating to the doughs. However, the rheology of a dough can also be inferred from its response to manipulation, shape retention, surface quality, the dynamics of oven expansion, and crumb morphology. The literature on gluten, GMP, and dough properties justifies this kind of interpretive reading, provided that it is declared qualitative and not as a direct measurement [1][2].

9.2. Four key rheological properties

In the protocol under examination, four properties matter above all:

1)Elasticity: the ability to recover at least part of the shape after deformation.
2)Extensibility: the ability to stretch without breaking.

3)Resistance to deformation: the ability to oppose gas pressure and manipulation without yielding too early.

4)Gas retention capacity: the most important integrated property, requiring continuity of the matrix and coordination of the surface skin.

This distinction is consistent with Wieser’s classical framework on the complementary function of gliadins and glutenins [1].

9.3. Rheological reading of Series I