Impact of redox agents on the extractability of gluten proteins during bread making

The gluten proteins gliadin and glutenin are important for dough and bread characteristics. In the present work, redox agents were used to impact gluten properties and to study gliadin-glutenin interactions in bread making. In control bread making, mixing increased the extractability of glutenin. The level of SDS-extractable glutenin decreased during fermentation and then further in the oven. The levels of extractable alpha- and gamma-gliadin also decreased during bread baking due to gliadin-glutenin polymerization. Neither oxidizing nor reducing agents had an impact on glutenin extractabilities after mixing. The redox additives did not affect omega-gliadin extractabilities during bread making due to their lack of cysteine residues. Potassium iodate (0.82-2.47 micromol/g of protein) and potassium bromate (1.07-3.17 micromol/g of protein) increased both alpha- and gamma-gliadin extractabilities during baking. Increasing concentrations of glutathione (1.15-3.45 micromol/g of protein) decreased levels of extractable alpha- and gamma-gliadins during baking. The work not only demonstrated that, during baking, glutenin and gliadin polymerize through heat-induced sulfhydryl-disulfide exchange reactions, but also demonstrated for the first time that oxidizing agents, besides their effect on dough rheology and hence bread volume, hinder gliadin-glutenin linking during baking, while glutathione increases the degree of covalent gliadin to glutenin linking.     

Determination of total folate in plant material by chemical conversion into para-aminobenzoic acid followed by high performance liquid chromatography combined with on-line postcolumn derivatization and fluorescence detection

A procedure involving chemical conversion of all forms of folate present in plant material into para-aminobenzoic acid (PABA) and a liquid chromatographic-fluorimetric determination with on-line postcolumn derivatization is reported. All folates are cleaved with liberation of PABA by hydrogen peroxide followed by acid hydrolysis using concentrated hydrochloric acid (37%) at 110 degrees C for 6 h. The reaction yield for individual folates conversion to PABA ranged from 44.4 to 97.3%. PABA could be determined sensitively by on-line postcolumn derivatization with fluorescamine, the detection limit for PABA being 3.02 nM. On the basis of this principle, a method for the determination of total folate in plant material, including a purification step on an affinity column, is presented, which offers a sufficient sensitivity and selectivity for routine analysis of total folate in natural samples. The total folate contents of tomatoes, carrots, white cabbage, and spinach were determined, and the results were quite comparable to the data reported. The recovery of PABA and the comparison of total folate analysis in spinach on different occasions (over 6 months) are also reported. The method is reliable, universal for all folates, including polyglutamate and monoglutamate forms, and eliminates the need for a deconjugation step and multiple conversion reactions.     

Impact of browning reactions and bran pigments on color of parboiled rice

Rice color changes from white to amber during parboiling (soaking and steaming). Color parameters indicated that, during soaking, yellow bran pigments leached out in the water. The levels of the Maillard precursors (i.e., reducing sugars (RS) and free alpha-amino nitrogen (FAN)) depended on soaking temperature and time: leaching of RS was compensated by enzymic formation for long soaking times (>60 min), while proteolytic activity was too low to compensate for FAN leaching. Rice soaking under nitrogen, oxygen, or ambient conditions and determination of polyphenol oxidase activity allowed us to conclude that the effect of enzymic color changes on the soaked rice color was rather small. Color measurements of brown and milled mildly, intermediately, and severely parboiled rice samples showed that both brown and milled rice samples were darker and more red and yellow after parboiling and that the effect depended on the severity of parboiling conditions. Furthermore, steaming affected the rice color more and in a way opposite to that observed in soaking. The changes in RS and the loss of FAN during parboiling suggested that Maillard type reactions occur during brown rice steaming. Analyses of furosine levels confirmed Maillard browning of outer bran layers and endosperm during steaming. The level of this Maillard indicator increased with the severity of parboiling conditions in both brown and milled parboiled rice. Measurements of the levels of bran pigments indicated that bran pigments diffuse into the endosperm during parboiling and contribute to the parboiled rice color.     

Baking Gradients Cause Heterogeneity in Starch and Proteins in Pound Cake

We investigated the impact of temperature and moisture gradients on starch gelatinization and egg denaturation, and on protein extractabilities during cake baking. Differences in crumb structure in the center, top, and bottom zones of cake as measured with X‐ray microfocus‐computed tomography were successfully related to the moment at which starch gelatinized and protein aggregated during baking, which stiffened the cell walls. The temperature in the top and bottom zones of cake increased faster than in the center of the cake due to facilitated heat transfer. This resulted in lower water availability in top and bottom zones, leading to incomplete gelatinization of starch after baking in these zones. In the top zone, extended starch gelatinization and protein polymerization led to later cell wall formation, resulting in a broader cell size distribution. The bottom zone of cake reached the highest temperatures during baking with more substantial starch gelatinization and egg denaturation within the first 25 min of baking. During the final 20 min of baking, little if any change in gelatinization enthalpy and protein extractability was found due to the very low water availability in this region. The bottom zone of the crumb showed a broader cell wall size distribution, which was associated with more collapse. All in all, the results illustrate that cake crumb is not a homogeneous material.     

Reaction kinetics of gliadin-glutenin cross-linking in model systems and in bread making

The gluten proteins gliadin and glutenin are important for wheat flour functionality in bread making, where, during baking, they polymerize through a heat-induced sulfhydryl-disulfide exchange mechanism. A model system was used to study the kinetics of this reaction. Thus, gluten was subjected to hydrothermal treatment with the rapid visco analyzer (RVA) with holding temperatures of 80, 90, and 95 degrees C. At these temperatures, omega-gliadin solubility did not change, but the solubilities of alpha- and gamma-gliadin in 60% ethanol decreased according to first-order reaction kinetics. All reaction rate constants increased with temperature. The activation energies for the heat-induced exchange reaction were 110 and 147 kJ/mol for alpha- and gamma-gliadin, respectively. Starch did not influence the reaction rates of the association of alpha- and gamma-gliadin with glutenin. During gluten-starch model bread baking, glutenin oxidized first, and when the internal crumb temperature reached 100 degrees C, alpha- and gamma-gliadin cross-linked to glutenin, again following first-order reaction kinetics. The experimental findings and similarities in temperature conditions and reaction kinetics suggest that the RVA system can be instrumental in understanding gluten behavior in concentrated food systems, such as bread making.     

Combined Effects of Endoxylanases and Reduced Water Levels in Pasta Production

The combined effects on pasta properties of 1) varying dosages of endoxylanases (EC 3.2.1.8) from Aspergillus aculeatus and Bacillus subtilis and 2) lower levels of water during pasta dough processing were studied. The A. aculeatus endoxylanase has high selectivity toward water‐extractable arabinoxylan (WE‐AX), whereas B. subtilis endoxylanase preferentially hydrolyzes water‐unextractable arabinoxylan (WU‐AX). Pasta was produced on a microscale (50.0 g) from the semolinas of both a strong (AC Navigator) and a moderately strong (AC Avonlea) durum wheat cultivar. The levels of added water in endoxylanase‐treated pastas were adjusted to obtain the same maximal farinograph consistencies as for the control pastas. The extruded pastas were dried with drying cycles at 40, 70, or 90°C. Apart from increasing levels of solubilized arabinoxylans, these treatments had little effect on the color, optimal cooking time, and firmness of the resulting pasta. High enzyme concentrations and low (40°C) drying temperature resulted in clearly or much less checked final products for the B. subtilis and A. aculeatus enzyme, respectively. Upon cooking, the enzymically formed low molecular weight arabinoxylans were retained better in the pasta strands than their equally low molecular weight arabinogalactan counterparts.     

Proteolytic activities in dormant rye (Secale cereale L.) grain.

Endoproteolytic, exoproteolytic, carboxypeptidase, aminopeptidase, and N-alpha-benzoyl-arginine-p-nitroanilide hydrolyzing activities were detected in 0.05 M sodium acetate buffer (pH 5.0) extracts of whole meal of the rye (Secale cereale L.) varieties Amando, Halo, and Humbolt. The proteolytic enzymes of Humbolt, the variety with the highest proteolytic activity, optimally hydrolyzed hemoglobin around pH 3.5 and 40-45 degrees C. In the different milling fractions of Humbolt, azocasein and hemoglobin hydrolytic activities were especially found in the bran and shorts. Proteolytic enzymes in the bran extract were concentrated in the 35-60% ammonium sulfate precipitate. Pepstatin A, an inhibitor of aspartic proteases, reduced approximately 88 and approximately 75% of the hemoglobin and azocasein hydrolyzing activities of this precipitate, respectively. Phenylmethanesulfonyl fluoride, an inhibitor of serine proteases, inhibited approximately 33% of both cited activities. Both rye and wheat storage proteins were degraded by Humbolt rye whole meal enzyme extract and the above-mentioned ammonium sulfate rye bran fraction in vitro. With the latter fraction digestion was more pronounced.