Effect of legume flours on baking characteristics of gluten-free bread

The objective of this work was to study the characteristics of four gluten-free bread formulations and the possibility of substituting soya protein with other legume proteins. Four bread recipes were prepared with chickpea flour, pea isolate, carob germ flour or soya flour. Carob germ flour batter structure was thicker compared with the other batters, probably due to the different protein behaviour and the residual gums present in carob germ flour. However, carob germ flour bread obtained the lowest specific volume values (2.51 cm³/g), while chickpea bread obtained the highest (3.26 cm³/g). Chickpea bread also showed the softest crumb. Confocal scanning-laser microscopy results showed a more compact microstructure in carob germ flour bread compared with soya and chickpea formulations. Chickpea bread exhibited the best physico-chemical characteristics and, in general, good sensory behaviour, indicating that it could be a promising alternative to soya protein.     

The relationship between rheological characteristics of gluten-free dough and the quality of biologically leavened bread

The rheological characteristics of gluten-free doughs and their effect on the quality of biologically leavened bread were studied in amaranth, chickpea, corn, millet, quinoa and rice flour. The rheological characteristics (resistance to extension R, extensibility E, R/E modulus, extension area, stress at the moment of dough rupture) were obtained by uniaxial dough deformation. Specific loaf volume of laboratory prepared gluten-free breads was in significant positive correlation with dough resistance (r = 0.86), dough extensibility (r = 0.98) and peak stress at the moment of dough rupture (r = 0.96). Even if the correlation between R/E modulus and the characteristics of loaf quality were not significant, the breads with the highest specific loaf volume were prepared from flours with R/E closer to the wheat check sample (18 N?mm-1). The results showed, in general, good baking flours exhibited stronger resistance to extension and greater extensibility, but differences found were not directly related to the results of baking tests.     

Improved viscoelastic zein–starch doughs for leavened gluten-free breads: Their rheology and microstructure

Gluten-free bread was prepared from commercial zein (20 g), maize starch (80 g), water (75 g), saccharose, NaCl and dry yeast by mixing above zein's glass transition temperature (T_g) at 40°C. Addition of hydroxypropyl methylcellulose (HPMC, 2 g) significantly improved quality, and the resulting bread resembled wheat bread having a regular, fine crumb grain, a round top and good aeration (specific volume 3.2 ml/g). In model studies, HPMC stabilized gas bubbles well. Additionally, laser scanning confocal microscopy (LSCM) revealed finer zein strands in the dough when HPMC was present, while dynamic oscillatory tests showed that HPMC rendered gluten-like hydrated zein above its T_g softer (i.e. |G*| was significantly lower). LSCM revealed that cooling below T_g alone did not destroy the zein strands; however, upon mechanical impact below T_g, they shattered into small pieces. When such dough was heated above T_g and then remixed, zein strands did not reform, and this dough lacked resistance in uniaxial extension tests. When within the breadmaking process, dough was cooled below T_g and subsequently reheated, breads had large void spaces under the crust. Likely, expanding gas bubbles broke zein strands below T_g resulting in structural weakness.     

Rheological properties and baking performance of rye dough as affected by transglutaminase

Since protein aggregation and formation of a continuous protein matrix in rye dough is very limited, an enzyme-induced protein aggregation method to improve the baking properties was investigated. The effects of microbial transglutaminase (TG) on the properties of rye dough were studied by rheological tests, confocal laser scanning microscopy (CSLM), standard-scale baking tests and crumb texture profile analysis. Addition of TG in the range of 0-4000 Ukg⁻¹ rye flour modified the rheological properties of rye flour dough, resulting in a progressive increase of the complex shear modulus (|G∗|) and in a decrease of the loss factor (tan δ) due to protein cross-linking or aggregation. CLSM image analysis illustrated a TG-induced increase of the size of rye protein complexes. Standard baking tests showed positive effects on loaf volume and crumb texture of rye bread with TG applied up to 500 Ukg⁻¹ rye flour. Higher levels of TG (500 U ≤ TG ≤ 4000 U) had detrimental effects on loaf volume. Increasing TG concentration resulted in an increase of crumb springiness and hardness. In conclusion, the results of this work demonstrated that TG can be used to improve the bread making performance of rye dough by creating a continuous protein network.     

Effect of the addition of extruded wheat flours on dough rheology and bread quality

Extruded wheat flours, due to their increased water absorption capacity, constitute an opportunity to increase bread output in bakery production. However extrusion may modify dough and bread characteristics. The aim of this study was to investigate the effect of the substitution of 5% of the wheat flour by extruded wheat flour (produced with different time-temperature extrusion treatments) on dough mixing, handling and fermentation behaviour and bread volume, shape, texture and colour. The RVA curves indicate that extrusion intensity increases with increasing temperature or water content. Water absorption capacity rises with increasing treatment intensity, but dough stability tends to decrease. Adding extruded flours decreases dough extensibility but increases tenacity and gas production. Differences in dough structure were observed on photomicrography, though there were no clear differences in bread quality. These results indicate that it is possible to obtain adequate dough and bread characteristics using dough with 5% extruded wheat flour.     

Effect of glucose oxidase,transglutaminase, and pentosanase on wheat proteins: Relationship with dough properties and bread-making quality

Glucose oxidase (Gox), transglutaminase (TG), and pentosanase (Pn) were investigated for their effect on bread quality. The changes introduced in wheat protein by the action of these enzymes were analysed to explain dough behaviour. Gox treatment decreased free sulphydryl groups (SHf), increased glutenin macropolymer contents, and modified the electrophoretic pattern of protein fractions. Gox modified mainly albumin, globulin, and glutenin, forming large protein aggregates. These modifications explained the high strength of the dough and the low bread specific volume of samples with Gox. TG treatment modified solubility in SDS of protein and decreased glutenin macropolymer content. However, it formed large protein aggregates. The new cross-linking bonds introduced by this enzyme were different to S–S bonds and, consequently, the dough was less extensible and showed high resistance. Pn treatment increased water soluble pentosan content. Moreover, in these samples a tendency to increase SHf content was observed. In addition, Pn increased protein solubility in isopropanol, which indicates that the reduction of pentosans size decreases steric impediment of insoluble pentosans, thus increasing interaction among protein and making their extraction easier. These changes at the microscopic level allowed explaining the formation of softer dough and the production of higher specific volume in breads with Pn.     

Volume, texture, and molecular mechanism behind the collapse of bread made with different levels of hard waxy wheat flours

Physico-chemical properties of bread baked by partially replacing normal wheat (Triticum aestivum L.) flour (15, 30, and 45%) with two hard waxy wheat flours were investigated. Substitution with waxy wheat flour resulted in higher loaf volume and softer loaves. However, substitution at >30% resulted in excessive post-bake shrinkage and a ‘key-hole’ shape with an open crumb structure. Bread crumb microstructure indicated a loss of starch granule rigidity and fusing of starch granules. The cells in the interior of the bread did not become gas-continuous and as a result, shrunk as the loaf cooled. Soluble starch content was significantly higher in bread crumb containing waxy wheat flour than in control bread. Debranching studies indicated that the soluble starch in bread made with 30-45% hard waxy wheat flour was mostly amylopectin. Incorporation of waxy wheat flour resulted in softer bread immediately after baking but did not retard staling upon storage.     

Agronomic characteristics, grain quality and flour rheology of 372 bread wheats in a worldwide core collection

A core collection of 372 accessions representative of worldwide hexaploid bread wheat diversity [Balfourier, F., Roussel, V., Strelchenko, P., Exbrayat-Vinson, F., Sourdille, P., Boutet, G., Koenig, J., Ravel, C., Mitrofanova, O., Beckert, M., Charmet, G., 2007. A worldwide bread wheat core collection arrayed in a 384-well plate. Theoretical and Applied Genetics 114, 1265–1275] was used to evaluate the available genetic diversity of agronomic and quality characteristics. The traits assessed during the vegetative period were date of ear-emergence, date of flowering, lodging, disease susceptibility and pre-harvest sprouting. Thousand kernel weight, test weight, grain hardness, grain protein content, pentosan viscosity and grain colour were also measured. The rheological properties of the derived white flours were estimated using mixograph and alveograph tests. For most of the traits, a wide phenotypic variation was observed across all the accessions. Several parameters (mixograph width parameters before and after peak time, alveograph dough tenacity and extensibility, near infrared measurements, like those for protein content, and absorbance measurements of palmitic acid and linoleic acid content) made it easier to discriminate between the cultivars. The largest ranges of variation were found in landraces and old cultivars rather than in more recent varieties. This is evidence that there is sufficient variability available for rare alleles, which have been eliminated in breeding modern varieties to be detected. Such a core collection will therefore be a useful resource for future genetic studies on wheat quality.     

Malt hydrolysates for gluten-free applications: Autolytic and proline endopeptidase assisted removal of prolamins from wheat,barley and rye

Cereal based products intended for gluten sensitive individuals, particularly to celiac disease patients, tend to have poor organoleptic qualities and they contain low levels of healthy whole grain compounds. Adding whole grain ingredients, such as malt hydrolysates, could compensate these defects provided that the ingredients are adequately free from toxic prolamin epitopes. Here we demonstrate that the level of toxic prolamin epitopes in the malt autolysates (wheat, barley, rye) were substantially lower than in the native malts but too high to allow “very low in gluten” labelling. To further eliminate the residual levels of toxic prolamin epitopes, a proline-specific endoprotease from Aspergillus niger was added to the malt autolysates. In the resulting malt hydrolysates (of wheat and rye but not barley), the prolamins were indeed greatly reduced and were below the very low gluten limit of 100 mg/kg. Malt hydrolysates with adequately low gluten levels may potentially be used as novel ingredients within gluten-free foods.     

Evaluation of heat damage, sugars, amylases and colour in breads from einkorn, durum and bread wheat flours

To limit nutritional losses and optimise bread processing, heat damage indices (furosine, glucosylisomaltol, hydroxymethylfurfural), sugars, α-amylase, β-amylase and colour were monitored during bread manufacturing from refined flour of three einkorn, three bread and one durum wheat samples. The heat damage indices increased only during the baking step. Furosine was significantly lower in einkorn (on average, 9.3 ± 5.33 and 25.3 ± 10.70 mg/100 g protein in crumb and crust, respectively) than in bread wheat (31.6 ± 3.05 and 115.6 ± 13.53) and durum wheat (36.2 ± 2.82 and 165.0 ± 3.17). Glucosylisomaltol and hydroxymethylfurfural were detected only in the crust, with lower levels in einkorn (on average, 2.3 ± 1.78 and 10.0 ± 7.79 mg/kg DM, respectively) than in bread wheat (13.1 ± 5.57 and 42.8 ± 10.64) and durum wheat (18.9 ± 1.11 and 57.2 ± 0.80). The different behaviour of einkorn was probably related to its moderate β-amylase activity, and thus the low maltose content of its dough. Colour was correlated with heat damage, as einkorn breads were lighter than the others.The results show that einkorn bread undergoes lower heat damage than analogous products from durum and bread wheat, thus probably better preserving its nutritional value.     

Effect of quinoa flour on gluten-free bread batter rheology and bread quality

A new gluten-free bread formulations composed of quinoa, buckwheat, rice flour and potato starch were developed in the present study. Rheological characteristics of the bread batter with increasing amount of quinoa were determined; storage (G′) and loss modulus (G″) values were also measured for investigation of viscoelastic properties. To evaluate the quality of breads; technological and physical (bake loss %, specific volume, texture, microstructure, color), chemical (protein, moisture, ash) and sensory properties were determined. All batter formulations independent of the quinoa amount exhibited pseudoplastic behavior, and G′ values were found to be higher than G″ values in expressing the solid like characteristics of the batter. Amount of quinoa flour addition did not present significant difference on bake loss%, specific volume and protein content (p>0.05); however, 25% quinoa flour bread displayed better results with its higher sensory scores and softer texture. Quinoa and buckwheat flour mixture therefore will be a good alternative for conventional gluten-free bread formulations.     

Improvement of dietary fiber,ferulic acid and calcium contents in pan bread enriched with nejayote food additive from white maize (Zea mays)

Nejayote is the wastewater from the alkaline-cooking of maize and its solids are rich in dietary fiber (45.3%), calcium (5.7%) and ferulic acid (219 mg/100 g). Nejayote solids were used to develop a food additive (NS) consisting of 80% nejayote solids and 20% gluten. NS was incorporated at 3, 6 or 9%, in wheat flour to increase the dietary fiber, calcium, phenolics and antioxidant capacity of breads. The addition of 9% NS did not affect overall baking performance and bread quality but increased dietary fiber up to 54% in composite breads. Moreover, enriched breads contained about 745 times more free ferulic acid and increased approximately 70% their antioxidant capacity. Two slices of bread (64 g) supplemented with 9% of NS provided 29% of the recommended calcium intake. Thus, the NS could be used as a value-added food-ingredient for the preparation of composite bakery-products with improved dietary fiber, calcium, nutraceuticals and antioxidant properties.     

Post-harvest characteristics of potato minitubers with different fresh weights and from different harvests. II. Losses during storage

Summary Storage losses were studied in minitubers of cvs Agria and Liseta, using five fresh weight classes (<0.50 g, 0.5–0.99 g, 1.00–1.99 g, 2.00–2.99 g, ≥3.00 g), and three successive harvests of the same plantlets. After each harvest, tubers were dired at room temperature (1 day), curred at 18 °C (13 days) and stored at 2 °C (540 days). Two kinds of storage losses were considered: (a) losses of entire tubers because of deterioration, and (b) fresh weight losses of the other tubers. Both kinds of losses were higher in cv. Liseta, in tubers with lower fresh weights and in tubers from the first harvest. Almost all minitubers ≥0.5 g from later harvests and from both cultivars survived storage for 1.5 years. Deterioration occurred mainly from 6 to 12 months of storage. Tubers which deteriorated during cold storage had already shown high weight losses during curing.     

Post-harvest characteristics of potato minitubers with different fresh weights and from different harvests. I. Dry-matter concentration and dormancy

Summary Dry-matter concentration and dormancy were studied in minitubers of cvs Agria and Liseta, using five fresh weight classes (<0.50 g, 0.50–0.99 g, 1.00–1.99 g, 2.00–2.99 g and ≥3.00 g) and three successive harvests of the same plantlets. The average dry-matter concentration increased with tuber weight for tubers from the second and third harvests. In minitubers ≥0.5 g, dry-matter concentration was higher in tubers from later harvests. The dormant period (days from harvest to 50% sprouting) was longer in minitubers with lower than higher weights, and longer in tubers from the first harvest than from later harvests. A cold-storage period of 6 weeks, starting 14 days after harvest, reduced the dormant period by an average of 11 days.     

Influence of storage on chemical,microbial and consumer acceptability of a milk-like product made from melon seeds

The storage stability of melon milk at room (30 ± 2 ^°C)and refrigeration (10 ± 2 ^°C) temperatures was determined by analyzing changes in the chemical, microbial andsensory properties of the milk stored for 7 days. Theresults showed that at both storage temperatures, solublesolids and pH of the milk decreased while titratableacidity increased with storage. The standard plate countsincreased appreciably while coliforms were absent in themilk with storage at both storage temperatures. The overallacceptability score of the melon milk dropped duringstorage, the drop being faster at 30 ± 2 ^°C than at 10 ± 2 ^°C. The milk samples stored at 30 ± 2 ^°C and 10 ± 2 ^°C were acceptable only within one and three days, respectively; thereafter, theywere unacceptable.     

Effects of homogenization pressure and stabilizers on some physical characteristics of a beverage prepared from partially defatted, roasted peanuts

A low-fat beverage with roasted peanut flavor was developed from peanuts. The potential milk substitute contained 11.8% total solids, 2.0% fat and 3.7% protein, and was whitish orange-yellow in color. The effects of stabilizing additives and homogenization pressure (13.8×10⁶ Pa or 41.4×10⁶ Pa) on selected physical characteristics were determined. Suspension stability was improved by addition of carrageenan [Benlacta CM61-B (CM), 0.02–0.04%] or a hydrogenated mono- and diglyceride [Emuldan HV52K (HV, 0.2–0.4%)]. Homogenizing at 41.4×10⁶ Pa increased viscosity of all samples except that of the controls but did not improve the suspension stability (top:bottom solids) of the beverage. The most viscous formulations (17.5±0.95 cps) were those containing 0.04% CM. Treatments yielding the best combination, compared to cow's milk values, of high suspension stability (0.5±0.03, where 1.0=maximum stability) and low viscosity (3.7±0.89 cps) were those containing 0.2% HV which were homogenized at 13.8×10⁶ Pa.     

Effect of ensiling field bean, field pea and common vetch in different proportions with whole-crop wheat using formic acid or an inoculant on fermentation characteristics

Whole‐crop field bean (FB), field pea (FP) and common vetch (CV) [155, 213 and 238 g dry matter (DM) kg^?1] were ensiled in 1·5 L laboratory silos with whole‐crop wheat as mixtures of 0, 0·25, 0·50, 0·75 and 1·00 of fresh weight (FW). Silages were ensiled (i) without additive, and (ii) with formic acid (FA) (4 L t^?1) or (iii) an inoculant (Lactobacillus plantarum, 10⁶ colony‐forming units g^?1 FW) as additives. The concentrations of water‐soluble carbohydrates in herbage of whole‐crop FB, FP, CV and wheat were 93, 157, 67 and 114 g kg^?1 DM and the buffering capacities were 588, 710, 755 and 429 mEq kg^?1 DM respectively. Field bean and FP silages were mainly well preserved with low pH values and moderate fermentation losses, except for FB‐only silage without additive which had a high butyric acid concentration. Common vetch silages had higher pH values and were less well fermented compared to the silages of the other legumes. For all legumes, FA reduced ammonia‐N concentrations more effectively compared to other additive treatments. In conclusion, in FB and FP silages the use of FA or an inoculant, as additives, ensured good preservation up to a proportion of legume in the herbage of 0·75. With all legume silages, and with those containing CV, only FA, as an additive, adequately restricted protein breakdown.     

Effects of two mixtures of perennial ryegrass cultivars with contrasting heading dates, and differing in spring-grazing frequency and silage harvest date, on characteristics of silage from first-cut swards

This experiment quantified the effects of: (i) heading date of perennial ryegrass, (ii) grazing frequency in spring and (iii) date of silage harvest, on the ensilability of herbages harvested for silage, and on the conservation and estimated nutritive value of the resultant silages. Replicated field plots with two perennial ryegrass mixtures (intermediate‐ and late‐heading cultivars) were subjected to three spring‐grazing regimes (no grazing, grazing in late March and grazing in both late March and late April) and were harvested on four first‐cut harvest dates between 20 May and 21 June. Herbage from each of the four replicates of these 24 treatments was precision‐chopped and ensiled unwilted and with no additive in laboratory silos. Herbage from the sward with the intermediate‐heading cultivar had a higher (P < 0·001) dry‐matter (DM) content and buffering capacity than that from the late‐heading cultivar, whereas water‐soluble carbohydrate concentrations increased (P < 0·001) with more frequent grazing in spring. Later harvesting enhanced herbage ensilability through an increased (P < 0·001) DM content and reduced (P < 0·001) buffering capacity and pH. Fermentation profiles of the silage were not markedly influenced by the cultivar mixture used but grazing in both late March and late April resulted in a more extensive fermentation with the acids produced increasingly dominated by lactic acid. The concentrations of acetic acid, and to a lesser extent, ethanol declined as silage harvest date was delayed. Overall, the relative effects of grass cultivar mixture were smaller than those of spring‐grazing treatment or silage‐harvesting date although on any given harvest date the herbage from the intermediate‐heading cultivar mixture was easier to preserve as silage than herbage from the late‐heading cultivar mixture. Delaying the harvesting of the late‐heading swards by 8 d removed the differences related to growth stage in buffering capacity, pH and DM content.     

Stomach Chitinase from Japanese Sardine Sardinops melanostictus: Purification,Characterization, and Molecular Cloning of Chitinase Isozymes with a Long Linker

Fish express two different chitinases, acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2), in the stomach. AFCase-1 and AFCase-2 have different degradation patterns, as fish efficiently degrade chitin ingested as food. For a comparison with the enzymatic properties and the primary structures of chitinase isozymes obtained previously from the stomach of demersal fish, in this study, we purified chitinase isozymes from the stomach of Japanese sardine Sardinops melanostictus, a surface fish that feeds on plankton, characterized the properties of these isozymes, and cloned the cDNAs encoding chitinases. We also predicted 3D structure models using the primary structures of S. melanostictus stomach chitinases. Two chitinase isozymes, SmeChiA (45 kDa) and SmeChiB (56 kDa), were purified from the stomach of S. melanostictus. Moreover, two cDNAs, SmeChi-1 encoding SmeChiA, and SmeChi-2 encoding SmeChiB were cloned. The linker regions of the deduced amino acid sequences of SmeChi-1 and SmeChi-2 (SmeChi-1 and SmeChi-2) are the longest among the fish stomach chitinases. In the cleavage pattern groups toward short substrates and the phylogenetic tree analysis, SmeChi-1 and SmeChi-2 were classified into AFCase-1 and AFCase-2, respectively. SmeChi-1 and SmeChi-2 had catalytic domains that consisted of a TIM-barrel (β/α)₈–fold structure and a deep substrate-binding cleft. This is the first study showing the 3D structure models of fish stomach chitinases.     

Antitumor Effects and Related Mechanisms of Penicitrinine A,a Novel Alkaloid with a Unique Spiro Skeleton from the Marine Fungus Penicillium citrinum

Penicitrinine A, a novel alkaloid with a unique spiro skeleton, was isolated from a marine-derived fungus Penicillium citrinum. In this study, the isolation, structure and biosynthetic pathway elucidation of the new compound were described. This new compound showed anti-proliferative activity on multiple tumor types. Among them, the human malignant melanoma cell A-375 was confirmed to be the most sensitive. Morphologic evaluation, apoptosis rate analysis, Western blot and real-time quantitative PCR (RT-qPCR) results showed penicitrinine A could significantly induce A-375 cell apoptosis by decreasing the expression of Bcl-2 and increasing the expression of Bax. Moreover, we investigated the anti-metastatic effects of penicitrinine A in A-375 cells by wound healing assay, trans-well assay, Western blot and RT-qPCR. The results showed penicitrinine A significantly suppressed metastatic activity of A-375 cells by regulating the expression of MMP-9 and its specific inhibitor TIMP-1. These findings suggested that penicitrinine A might serve as a potential antitumor agent, which could inhibit the proliferation and metastasis of tumor cells.