Thermal denaturation and gelation of vicilin-rich protein isolates from three Phaseolus legumes: A comparative study

The thermal denaturation and gelation properties of vicilin-rich protein isolates from red bean, red kidney bean and mung beans (further denoted as RPI, KPI and MPI) were investigated by differential scanning calorimetry (DSC) and dynamic oscillatory measurements. The relation between the properties of these proteins to their free sulphydryl (SH)/disulfide bond (SS) contents was also evaluated. DSC analyses showed that many DSC characteristics of major endothermic peak (corresponding to vicilin component), including its denaturation temperature (T_d), enthalpy changes (ΔH) and width at half-peak height (ΔT_1/2), significantly varied with the type of protein isolates. Furthermore, the heat-induced gelation, including onset of gelation and the development of mechanical moduli, was also dependent on the type of protein isolates. The thermal denaturation of these proteins was nearly unaffected by the presence of reducing agent dithiothreitol (DTT), while the presence of DTT weakened the gel formation. The T_d of vicilin components and the mechanical moduli of corresponding formed gels were positively related to their SS contents. Additionally, the formed gels were thermo-irreversible, and heat pretreatment (carried out at temperatures higher than or close to the T_d of these vicilins) could improve the gel network formation. These results confirm that vicilin-rich protein isolates from various legumes show different patterns of thermal denaturation and gelation, and these properties are to a great extent related to their SH and/or SS contents.     

Characterization,amino acid composition and in vitro digestibility of hemp (Cannabis sativa L.) proteins

The protein constituents and thermal properties of hemp (Cannabis sativa L.) protein isolate (HPI) as well as 11S- and 7S-rich HPIs (HPI-11S and HPI-7S) were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and different scanning calorimetry (DSC), and their amino acid composition and in vitro digestibility were also evaluated, as compared to soy protein isolate (SPI). SDS-PAGE analysis showed that the edestin (consisting of acidic and basic subunits, AS and BS) was the main protein component for HPI and HPI-11S, while HPI-7S was composed of the BS of edestin and a subunit of about 4.8 kDa. DSC analysis characterized thermal transition of the edestin component and the possible present form of different subunits. Except lysine and sulfur-containing amino acids, the essential amino acids of various HPIs met the suggested requirements of FAO/WHO for 2–5 year old infants. The proportion of essential amino acids to the total amino acids (E/T) for HPI (as well as HPI-11S) was significantly higher than that of SPI. In an in vitro digestion model, various protein constituents of various HPIs were much easily digested by pepsin plus trypsin, to release oligo-peptides with molecular weight less than 10.0 kDa (under reduced condition). Only after pepsin digestion, in vitro digestibility of HPIs was comparable to that of SPI, however after pepsin plus trypsin digestion, the digestibility (88–91%) was significantly higher than that (71%) of SPI (P < 0.05). These results suggest that the protein isolates from hempseed are much more nutritional in amino acid nutrition and easily digestible than SPI, and can be utilized as a good source of protein nutrition for human consumption.     

Physicochemical, functional and structural properties of vicilin-rich protein isolates from three Phaseolus legumes: Effect of heat treatment

The physicochemical, functional and structural properties of vicilin-rich protein isolates from kidney, red and mung beans (KPI, RPI and MPI) were investigated, and the influences of heating (at 95 °C for 30 min) were also evaluated and compared. In the untreated samples, the thermal stability, free SH contents and surface hydrophobicity were different. The differences seemed to be closely related to the differences in extent of aggregation and/or tertiary and secondary conformational structures among proteins. The heating resulted in extensive denaturation of the protein, significant decreases in free SH groups and increases in surface hydrophobicity, but to a varying extent, depending on the type of protein isolates. The protein solubility and emulsifying activities were significantly improved by the heating. The tertiary and secondary structures of these proteins were also to a various extent affected. The conformational structures of proteins in RPI were most flexible, and susceptible to the heating, while the proteins in KPI were most heat-stable in structures. The results clearly indicated close relationships between functional properties of these vicilin-rich protein isolates and their conformational structures.     

Effect of citric acid deamidation on in vitro digestibility and antioxidant properties of wheat gluten

The effects of citric acid deamidation on the physiochemical properties of wheat gluten were investigated. In vitro digestion was carried out to determine changes of molecular weight distribution, amino acids composition and antioxidant efficacy of wheat gluten hydrolysates. Results indicated that citric acid deamidation significantly increased gluten solubility and surface hydrophobicity, at a neutral pH. Deamidation induced molecular weight distribution change of gluten with little proteolysis. Results from FTIR indicated that the α-helix and β-turn of deamidated gluten increased accompanied by a decrease of the β-sheet structure. After deamidation, in vitro pepsin digestibility of wheat gluten decreased, while in vitro pancreatin digestibility increased. The oxygen radical absorbance capacity (ORAC) activity of the in vitro digests decreased with increase of deamidation time. The high Lys and total essential AAs amounts in the final digests suggested that the nutritional values of wheat gluten after deamidation might be enhanced.     

Properties of cast films of vicilin-rich protein isolates from Phaseolus legumes: Influence of heat curing

The mechanical, hydrophobic and thermal properties of cast films of vicilin-rich protein isolates from three Phaseolus legumes were investigated and compared to that of soy protein isolate (SPI). The influence of heat curing at 85 °C on the properties of these films was characterized. The films of vicilin-rich protein isolates exhibited much less mechanical strength (TS) and elongation at break (EB) and similar film surface hydrophobicity, as compared with those of the SPI film. The heating remarkably improved the TS of these films, and the extent of the improvement much higher than that of the SPI film, while the EB was only slightly affected. The thermal properties of these protein isolate films were variable, and much distinctly affected by the heating of the films, to a various extent, depending on the type of protein isolates. Protein solubility analyses indicated strengthened hydrogen and hydrophobic bondings of these films by the heating. The results suggest that those vicilin-rich protein isolates have good potential to form cast films with mechanical strength comparable to that of SPI film.     

Physicochemical changes of myofibrillar proteins during processing of Cantonese sausage in relation to their aggregation behaviour and in vitro digestibility

The physicochemical changes of myofibrillar proteins, especially oxidation behaviour, were measured to determine their mechanism of action on in vitro protein digestibility during Cantonese sausage processing. The results indicated that the carbonyl level significantly increased (p < 0.05) during the process. The SH group level decreased, while S–S group level increased gradually. Protein aggregation was induced by oxidation and heat treatment. Result from Fourier transform infrared (FTIR) spectroscopy confirmed protein aggregation occurred. The analysis of in vitro digestibility showed a highly significant (p < 0.05) correlation between pepsin activity and carbonyl group formation, S–S group level, protein surface hydrophobicity, D_4,3. A negative and highly significant correlation between trypsin, α-chymotrypsin activity and carbonyl group formation was measured, while no significant correlation with S–S groups, protein surface hydrophobicity, D_4,3 was observed. It indicated that not only protein oxidation and aggregation but also degradation by pepsin would influence proteolysis with trypsin and α-chymotrypsin.     

Microencapsulation properties of protein isolates from three selected Phaseolus legumes in comparison with soy protein isolate

The spray-drying microencapsulation properties of protein isolates from three selected Phaseolus legumes (kidney, red and mung beans; KPI, RPI and MPI), at a specific concentration (6 g/100 mL) and oil/protein ratio (1:1, w/w) were compared with soy protein isolate (SPI). The oil retention efficiency (RE), redispersion and dissolution behavior, as well as microstructure of the spray-dried powders were characterized. The influence of storage at 75% relative humidity for 7 days on these characteristics was also evaluated. The results indicated that the microencapsulation properties (except RE in the KPI case) of the three protein isolates were considerably poorer than SPI, though their emulsifying ability was even superior. The microencapsulating properties of these protein isolates were largely associated with their interfacial properties, especially the interfacial protein concentration. Among the three protein isolates, the spray-dried powders with KPI exhibited highest RE but least redispersion and/or dissolution behavior. The storage resulted in a severe loss of RE and ability to be redispersed and/or dissoluted, with much higher extent observed for the KPI powder. These results suggest that appropriate modifications, especially in interfacial properties, should be conducted on these proteins to warrant their application as wall materials in spray-drying microencapsulation.     

A study of the in vitro protein digestibility of indica and japonica cultivars

Protein digestibility was determined in 18 indica and japonica raw milled rices using an in vitro pH-drop method with three- or four-enzyme system. Similar protein digestibility was found between indica and japonica rices, which is in agreement with the in vivo digestibilities in human. Cooking improved protein digestibility in the four-enzyme assay, while reducing agents exhibited apparent inhibition in multienzyme digestibility of indica and japonica rices. A significant correlation was detected between protein content and the estimates of digestibility, whereas no significant correlation was found between amylose content and digestibility estimates. SDS–PAGE analysis showed a significant difference in the degradation extent of prolamin between multienzyme and pepsin digestion, which might contribute to the inconsistence between results of this study and previous findings that in vitro protein digestibility of japonica rice was higher than that of indica rice. In addition, our results supported the previous report that waxy gene product level is not a major determinant of protein digestibility in milled rice.     

Physicochemical, in vitro digestibility and functional properties of carboxymethyl rice starch cross-linked with epichlorohydrin

Cross-linked carboxymethyl rice starches (CL-CMRSs) were prepared from reactions between native rice starch and varied concentrations (0.1–15% w/w, M-0.1 to M-15) of epichlorohydrin (ECH) in a simultaneous carboxymethylation-crosslinking reaction setup using methanol as the solvent. The degree of carboxymethyl substitution was between 0.24 and 0.28, while apparent amylose contents were lowered due to modification. SEM images showed minor change on the granule surface, while XRD profiles indicated slight loss of crystallinity. DSC thermograms revealed no transition peak in all treated samples. The water uptake (WU), swelling volume (SV) and free swelling capacity (FSC) of CL-CMRSs increased significantly as a result of the modification, while swelling of CMRSs cross-linked with 2% (M-2) and 3% (M-3) ECH yielded FSC values and WU values that were much greater than those of native starches and were comparable to that of Explotab®. All modified starch samples showed increased amount of rapidly digestible starch (RDS), while cross-linking with 5–7.5% ECH raised the resistant starch (RS) content, compared to native starch. M-2 also showed promising results in tablet disintegration test. ECH–CL–CMRSs could potentially be used as an excipient in pharmaceutical and food/food supplement products.     

Effect of soaking and hydrothermal processing methods on the levels of antinutrients and in vitro protein digestibility of Bauhinia purpurea L. seeds

The effects of various domestic processing methods such as soaking, cooking and autoclaving on the levels of certain antinutritional factors and in vitro protein digestibility of seeds of Bauhinia purpurea L., an underutilised legume collected from South India, were investigated. The raw seeds were found to contain antinutritional factors like total free phenolics (2.75 g/100 g), tannins (2.35 g/100 g), phytic acid (692 mg/100 g) and flatulence factors, raffinose (0.54 g/100 g), stachyose (1.17 g/100 g) and verbascose (0.95 g/100 g). Soaking the seeds in distilled water caused maximum reduction in the phytic acid content (37%), whereas soaking in NaHCO₃ solution reduced significant levels of phenolics and tannins (72% and 78%, respectively). A reduction in the levels of oligosaccharides (raffinose by 63%, stachyose by 42% and verbascose by 79%) was observed during cooking. Of the attempted treatments, autoclaving appeared to be most effective in reducing levels of all the investigated antinutrients, except phytic acid, and also improved the in vitro protein digestibility of B. purpurea seeds.     

Effect of heat,rutin and disulfide bond reduction on in vitro pepsin digestibility of Chinese tartary buckwheat protein fractions

Protein fractions (albumin, globulin, prolamin and glutelin) were extracted from defatted tartary buckwheat flour. The in vitro pepsin digestibilities of the four protein fractions were different, and albumin was more susceptible to pepsin hydrolysis. The native structure of the four protein fractions may be destroyed by heat treatment, and the digestibilities were all improved significantly (P < 0.05). Adding rutin to the digestion mixture of the four fractions did not cause a decrease in pepsin digestibility, although it did cause a significant increase in certain instances (P < 0.05). Treatment with β-mercaptoethanol (2-ME) only caused a higher initial proteolysis rate and did not increase the final digestibility distinctly except for prolamin. After pepsin digestion, the remaining proteins of unhydrolyzed albumin, globulin, prolamin and glutelin (untreated) shared some similarities. They also exhibited a minor band at 20,000 Da and a broad band at 10,000–14,000 Da.     

Comparison of in vitro digestion characteristics and antioxidant activity of hot- and cold-pressed peanut meals

Due to the poor protein solubility, hot-pressed peanut meal (HPM) has less value than cold-pressed peanut meal (CPM) in the food industry. The objective of this study was to determine whether the denatured proteins in HPM were suitable for hydrolysis by digestive enzymes. The hydrolysis characteristics and antioxidant activity of HPM and CPM during in vitro digestion were compared. The results showed that HPM was hydrolysed more extensively than CPM. There were more free amino acids and small peptides with MW < 5 kDa in HPM hydrolysates. In addition, HPM hydrolysates displayed stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,20-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging activities, ORAC value and reducing power than CPM hydrolysates, which could be due to the higher content of small peptides, antioxidative amino acids and melanoidins in HPM hydrolysates. The above results indicated that HPM was a potential source of protein supplement for human consumption.     

Relationship between the structure,physicochemical properties and in vitro digestibility of rice starches with different amylose contents

The in vitro digestibility and molecular and crystalline structures of rice starches (Long-grain, Arborio, Calrose, and Glutinous) differing in amylose content were investigated and the relationship between the structure and in vitro digestibility of starch was studied. Long-grain showed the highest amylose content (27.2%), whereas Glutinous showed the lowest amylose content (4.2%). Long-grain had the highest average amylopectin branch chain length (18.8) and proportion (8.7%) of long branch chains (DP ≥ 37), and the lowest proportion (26.9%) of short branch chains (DP 6–12). Among the non-waxy rice starches (Long-grain, Arborio, and Calrose), Calrose had the lowest average chain length (17.7) and the lowest proportion (7.1%) of long branch chains (DP ≥ 37). The relative crystallinity of rice starch followed the order: Glutinous (33.5%) > Calrose (31.4%) > Arborio (31.0%) > Long-grain (29.9%). Long-grain had the highest gelatinization temperature and the lowest gelatinization temperature range, whereas Glutinous showed the highest gelatinization temperature range and gelatinization enthalpy. Arborio had the highest melting enthalpy for amylose–lipid complex among the tested rice starches. Pasting temperature, setback, and final viscosity increased with increasing amylose content, whereas the peak viscosity and breakdown showed negative correlations with amylose content. The rapidly digestible starch (RDS) content of the tested rice starches followed the order: Glutinous (71.4%) > Calrose (52.2%) > Arborio (48.4%) > Long-grain (39.4%). Contrary to this, the slowly digestible starch (SDS) and resistant starch (RS) contents showed an opposite trend compared to RDS. Digestibility (RDS, SDS, and RS) of the rice starches was significantly correlated (p ≤ 0.05) with amylose content, proportions of DP 6–12 and DP 13–24, relative crystallinity, intensity ratio (of 1047 cm⁻¹ to 1022 cm⁻¹ from Fourier transform infrared spectroscopy), swelling factor, amylose leaching, onset temperature of gelatinization, gelatinization temperature range, gelatinization enthalpy, pasting temperature, peak viscosity, breakdown, setback, and final viscosity.     

Isolation, solubility and in vitro hydrolysis of chickpea vicilin-like protein

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4-67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion.     

A comparative study of tocopherols composition and antioxidant properties of in vivo and in vitro ectomycorrhizal fungi

In aerobic organisms, the free radicals are constantly being produced during the normal cellular metabolism. The antioxidant properties of many organisms and particularly of wild mushrooms with their content in antioxidant compounds such as tocopherols, can detoxify potentially damaging forms of activated oxygen. Herein, a comparative study of tocopherols composition and antioxidant properties of in vivo (fruiting bodies) and in vitro (mycelia) ectomycorrhizal fungi: Paxillus involutus and Pisolithus arhizus. Tocopherols were determined by high performance liquid chromatography (HPLC) coupled to a fluorescence detector. The antioxidant properties were studied in terms of DPPH radical-scavenging activity, reducing power and inhibition of β-carotene bleaching. Fruiting bodies revealed the highest antioxidant properties, including scavenging effects on free radicals (EC₅₀ = 0.61 and 0.56 mg/ml) and inhibition of lipid peroxidation capacity (EC₅₀ = 0.40 and 0.24 mg/ml for P. involutus and P. arhizus, respectively), than mycelia produced in vitro cultures. Nevertheless, mycelia revealed higher levels of total tocopherols than fruiting bodies, and particularly P. arhizus mycelium proved to be a powerful source of γ-tocopherol (154.39 μg/g dry weight).     

Chemical composition,antinutritional constituents,precaecal crude protein and amino acid digestibility in three unconventional tropical legumes in broilers

Three tropical crop seeds, namely Enterolobium cyclocarpum, Mucuna pruriens and Centrosema pubescens, were assessed for their contents of chemical constituents, antinutritional factors (ANFs), crude protein (CP) and amino acid (AA) digestibility. Their meals were incorporated into broiler diets in place of maize starch at two levels (150 and 300 g kg^?1). Four hundred and twenty 1‐day‐old broilers were randomly assigned to seven experimental diets on a weight basis and used to determine CP and AA digestibility in the diets and seeds, and performance of the birds. The CP contents were 237, 278 and 252 g kg^?1 DM in E. cyclocarpum, M. pruriens and C. pubescens, respectively. Compared to the basal diet, the digestibility of CP and AAs in the diets and seeds was significantly (p < 0.05) reduced at higher level of inclusion of E. cyclocarpum and M. pruriens but increased at higher C. pubescens inclusion. Digestibility of arginine, threonine and lysine in the C. pubescens meal diets was similar to that in the basal diet. Digestibility values of CP and AAs were least in E. cyclocarpum. Aspartic acid was the least and glutamic acid the most digested AA in all the seeds. Among the essential AAs, threonine was the least digested AA, while leucine (in E. cyclocarpum and M. pruriens) and lysine (in C. pubescens) had the highest digestibility with values of 65, 73 and 72%, respectively. Feed intake, body weights and weight gain in the birds were significantly (P < 0.05) reduced in the E. cyclocarpum and M. pruriens diets but increased in the C. pubescens diets compared to the basal diet. Results of the study suggest that, based on the CP content and AA profiles in the seeds, they have potential for poultry feeding. The presence of ANFs, especially in E. cyclocarpum and M. pruriens, however, could be a major limitation in their use in poultry feeding. Copyright © 2006 Society of Chemical Industry     

Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L.

Coriandrum sativum L. is a source of a variety of polyphenols and other phytochemicals, related to its high antioxidant activity and to its use for indigestion, rheumatism, and prevention of lipid peroxidation damage. Plant cell cultures are a means to study or to produce some active metabolites, such as polyphenols. This technique was applied to the investigation of coriander, and a detailed analysis of individual polyphenols in vivo and in vitro grown samples was performed. The in vivo vegetative parts showed quercetin derivatives as the main flavonoids and quercetin-3-O-rutinoside (3296 mg/kg dw) was the main polyphenol found in this part of coriander. The fruits revealed only phenolic acids and derivatives, caffeoyl N-tryptophan hexoside (45.33 mg/kg dw) being the most abundant phenolic derivative. In vitro samples also gave a high diversity of polyphenols, being C-glycosylated apigenin (2983 mg/kg dw) the main compound. Anthocyanins were only found in clone A, which was certainly related to its purple pigmentation, and peonidin-3-O-feruloylglucoside-5-O-glucoside was the major anthocyanin found (1.70 μg/kg dw). In vitro culture can be used to explore new industrial, pharmaceutical, and medicinal potentialities, such as the production of secondary metabolites like flavonoids.     

Effect of glycation on protein structure,amino acid composition and digestibility of silver carp mince

This study was to investigate the effects of glycation on the quality of silver carp mince (SCM) proteins. The browning index (BI), sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), Fourier transform infrared spectroscopy (FTIR), total amino acid composition, chemical score, protein digestibility and reductive power of digested products were determined. The correlation between the degree of glycation, protein structure and protein nutritional quality was analysed. The results showed that the BI value of SCM-G increased with the increase in heating time. Myosin heavy chain (MHC), myosin light chain (MLC), tropomyosin and actin were the main protein reactants for glycation. Glycation led to the expansion of the secondary structure of SCM proteins. Glycation reduced lysine and cysteine content but did not change the first limiting amino acid (Val) in SCM. The essential amino acid content of glycated SCM remained consistent with the amino acid nutritional requirements for children and above recommended by FAO/WHO. In addition, glycation did not change protein digestibility but significantly increased the reducing power of digested products with increasing degree of glycation. These results provided a theoretical basis for the application of glycation in food processing industry and the development of aquatic products with high nutritional value.