Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates

Antioxidative properties of proteins from barley and rice bran and their hydrolysates were examined. Three major hordein fractions of barley, B, C and D hordeins, were partially purified by gel filtration. Albumin, globulin, prolamin and glutelin fractions of rice bran were fractioned by the Osborne method. Hydrolysates of these protein fractions were prepared by digesting with pepsin followed by trypsin. Antioxidant properties in terms of antioxidative activity against linoleic acid peroxidation and reducing activity without the lipid adjuvant were investigated. The globulin fraction from rice bran protein revealed the strongest antioxidative activity throughout the incubation time of 7 days (p ≤ 0.05). The albumin fraction of rice bran protein showed the highest reducing activity (6964 μmol of Fe²⁺) followed by globulin, prolamin, glutelin and hordein fractions with activities of 2904, 2017, 1809 and 1333 μmol of Fe²⁺, respectively (p ≤ 0.05). Partially purified C hordein exhibited the highest reducing activity compared with B and D hordeins. Protein hydrolysates obtained after digestion with pepsin and trypsin exhibited much greater antioxidative, as well as reducing, activities than those from before digestion.     

Characteristics of enzymatic hydrolysis of thermal-treated wheat gluten

Effect of thermal treatment at 50–90 °C on wheat gluten hydrolysis by papain was evaluated in this study. Thermal treatment decreased the amount of sodium dodecyl sulfate (SDS) extractable protein. The treatments at 80 and 90 °C had a strong impact on protein extractability. Thermal treatment for 30 min resulted in a significant reduction in SDS extractable glutenin level in wheat gluten. A significant drop in free sulphydryl level was found in wheat gluten treated at 70 °C for 30 min. It indicated that cross-linking of glutenin through S–S occurred during thermal treatment. The treatments at 70–90 °C led to significant decreases in soluble and nitrogen level, while significant increases in peptide nitrogen amount in the hydrolysates from treated gluten were found. A time-dependent effect was observed for the changes in soluble forms of nitrogen and PN. Thermal treatment resulted in molecular mass distribution change according to gel permeation chromatography analysis. Thermal treatment significantly increased the amount of fractions with molecular mass beyond 10 K (67.2%) in the hydrolysates and greatly decreased the amounts of fractions with MM of 10–5 K and below 5 K in hydrolysates.     

Effect of extrusion treatment on enzymatic hydrolysis of wheat bran

Although, wheat bran is a good source of dietary fibre (DF), it has certain disadvantages due relatively lower levels of soluble DF. Therefore, in the current study, the effect of extrusion treatment on microstructure and enzymatic hydrolysis of wheat bran was investigated. Extrusion treatment increased fibre solubility at all process conditions and the screw speed was found to be the most effective parameter. Physicochemical properties of bran were affected from extrusion treatments. While the water-binding capacities of extruded brans were lower than that of non-extruded wheat bran, their water solubilities were higher. Enzymatic hydrolysis increased the soluble DF content of the bran samples as compared to those of respective samples at the beginning of incubation. The results showed that extrusion treatment can be used to disrupt the wheat bran microstructure and thus to increase the soluble fibre content. Enzymatic hydrolysis can also be used for increasing solubility further. The outcomes of this study can be utilized for improving the technological functionality of cereal fibres to develop high fibre ingredients for food applications.     

Effects of washing, milling and loading enzymes on the enzymatic hydrolysis of a steam pretreated sugarcane bagasse

Enzymatic hydrolysis of steam pretreated sugarcane bagasse was performed to investigate the production of glucose and xylose. A blend of industrial enzymes (Novozymes) was used. The enzymes were used either without having been washed or having been washed with water or 1% aqueous NaOH solution. The influence of the size of sugarcane bagasse and the proportion of the enzymatic cocktail, which was composed of endoglucanases/cellobiohydrolases (Celluclast 1.5L) and β-glucosidase (Novozym 188) cellulases mixtures, was investigated. The tests were performed at a temperature of 50 °C and at a pH of 4.8 during a period of 72 h. The assays that were conducted with a pretreated sugarcane bagasse that was washed with a 1% aqueous NaOH solution without milling led to the largest amounts of glucose after 72 h, which was independent of the proportions of enzymes used in experiments with the smallest amounts of enzymes. The largest amount of xylose was obtained with a pretreated sugarcane bagasse that was not washed.     

Influence of high solid concentrations on enzymatic wheat gluten hydrolysis and resulting functional properties

Enzymatic hydrolysis at increased solid concentrations is beneficial with regard to energy and water consumption. This study examines the influence of the solid concentration on the enzymatic hydrolysis of wheat gluten and the resulting functional properties of the hydrolysate. Wheat gluten was mildly hydrolyzed at a solid concentration varying from 10% to 60% to degrees of hydrolysis (DH%) ranging from 3.2% to 10.2%. The gluten was susceptible to hydrolysis at all solid concentrations but the hydrolysis rate was influenced by increasing solid concentrations. Size-exclusion high-performance liquid chromatography revealed an increase in the ratio of peptides with a molecular mass >25 kDa for solid concentrations of 40% and 60%. The water solubility increased on hydrolysis and was independent of the solid concentration during proteolysis. The foam stability was not influenced by the solid concentration at low DH%. At DH% higher than 8%, high solid concentrations increased the foam stability, which might be related to the presence of more peptides with a molecular mass >25 kDa. In addition, we found increased reactor productivity. The results show the potential of hydrolyzing wheat gluten at high solid concentrations, which could lead to large savings for water and energy when applied industrially.     

ABTS and DPPH methods as a tool for studying antioxidant capacity of spring barley and malt

Many analytical methods for antioxidant determination in foodstuffs and raw materials based on various principles have been published so far. However, not all of them are applicable to barley and malt. The results of total antioxidant capacity (TEAC) of barley and malt obtained with methods based on ability to eliminate radicals of 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) were compared and statistically evaluated. The average TEAC of barley determined using ABTS and DPPH ranged from 2.1 to 2.5 μmol g⁻¹ and from 1.2 to 1.7 μmol g⁻¹, respectively. The TEAC ranges in malt were 2.7–3.0 μmol g⁻¹ (ABTS) and 1.8–2.6 μmol g⁻¹ (DPPH). TEAC of barley and malt were affected by the weather conditions (year), variety and application of Zn²⁺ fertilizer. The ABTS and the DPPH methods represent an effective tool for the assessment of antioxidant capacity of spring barley and malt and the factors having an impact on it.     

Characterization of Amaranthus hypochondriacus seed protein fractions,and their antioxidant activity after hydrolysis with lactic acid bacteria

The objective of this work was to obtain and characterize five Amaranthus hypochondriacus seed protein fractions followed by the determination of antioxidant activities of hydrolysates produced with two Lactobacillus species. Molecular weights of polypeptides (SDS-PAGE) in fractions were 14–97 kDa and those identified by LC-MS/MS include agglutinin, trypsin inhibitor, and cystatin. Each fraction was hydrolyzed with Lactobacillus helveticus and Lactobacillus plantarum at their exponential (4 and 8 h) and stationary (14 and 24 h) growth phases. The free amino group content in hydrolysates produced with L. helveticus was up to 5.15% compared to 4.79% in those obtained with L. plantarum. In the antioxidant tests, higher ROO• scavenging activities were obtained for hydrolysates from albumins (1037 ± 80 μM TE/mL) and glutelins (1793 ± 168 μM TE/mL) fractions produced with L. helveticus and L. plantarum, respectively. Scavenging activities of •OH radicals were higher for 11S-G hydrolysates with both Lactobacillus at the stationary phase.     

Fate of polyphenols and antioxidant activity of barley throughout malting and brewing

Phenolic contents of barley and malt extracts and their corresponding antioxidant activities were investigated using a chromatographic online antioxidant detection system. Ethyl acetate extracts of barley and malt were separated using reverse phase HPLC and compounds eluting from the column were submitted to two UV–visible detections: one for the phenolic compounds; and the other for the reduced form of the radical cation 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS^•+) after the compounds were allowed to react online with it. Prodelphinidin B3 and procyanidin B3 were identified as two major contributors in the antioxidant activity of barley, in addition to catechin. Malting had a dramatic impact on these three compounds by resulting in a sharp decrease in their detected amounts and the associated antioxidant activities. Two other antioxidants, ferulic and sinapic acids, showed a better ability to withstand not only malting but also brewing steps. As for the overall phenolic content and antioxidant capacity, the study showed that malting allowed a better release and/or extraction of phenolic compounds, while the first brewing step caused the most significant damage by drastically decreasing the total polyphenols and their activity. Hopping however did not significantly affect neither the phenolic content nor the antioxidant activity.     

Composition and molecular structure of polysaccharides released from barley endosperm cell walls by sequential extraction with water,malt enzymes,and alkali

Isolated and purified endosperm cell walls (CW), used in this study, were derived from a Canadian malting barley variety, AC Metcalfe, grown in three different environments in Canada in 2003, and varying in grain protein and β-glucan contents, as well as in grain hardness. The CW were initially extracted with water at 45 °C and subsequently digested with barley malt crude enzyme extract resulting in two fractions designated CW-WE45 and CW-MD, respectively. The remaining non-digested cell wall material (CW_ND) was further fractionated by sequential extraction with water at 95 °C (CW_ND-WE95), saturated barium hydroxide (CW_ND-BaE), and 1 N sodium hydroxide (CW_ND-NaE) at 25 °C. Composition and molecular structure analyses were carried out for all fractions including the remaining cell wall residue (CW_RES). Extraction of CW with water followed by digestion with malt crude enzyme extract solubilized the majority of β-glucans (∼55–70%) and glucomannans (∼60–80%) but only a small portion of arabinoxylans (∼20–30%) present in the intact CW. The CW-WE45 and CW_ND-WE95 fractions consisted mostly of β-glucans exhibiting high average molecular weights (M_w) (2–3 × 10⁶), whereas the CW_ND-BaE consisted mainly of arabinoxylans with M_w about 1–1.5 × 10⁶. The CW_ND-NaE contained almost equal amounts of β-glucans and arabinoxylans and a small amount of glucomannans, whereas the CW_RES contained approximately equal proportions of β-glucans, arabinoxylans and glucomannans. β-Glucans in CW_ND-WE95, CW_ND-NaE, and CW_RES exhibited a higher ratio of 3-O-β-d-cellobiosyl-d-glucose to 3-O-β-d-cellotriosyl-d-glucose (DP3/DP4) compared to β-glucans in CW-WE45 and CW-MD. β-Glucans in CW_ND-NaE showed the highest level of long cellulosic oligosaccharides with DP ≥ 5, whereas those in the CW_RES had the highest DP3/DP4 ratio. The CW-MD was fractionated by ultrafiltration into high (CW-MD_HMW) and low-molecular weight (CW-MD_LMW) sub-fractions, with weight-average M_w of ∼150–350 × 10³ and <10 × 10³, respectively, as confirmed by size-exclusion chromatography. The monosaccharide composition of the sub-fractions indicated a more extended enzymic degradation of β-glucans and glucomannans than arabinoxylans. Some differences in composition and molecular structure of the cell wall constituents among the three barley samples were related to their solubility and enzymic digestibility.     

The production and characterisation of Hor 3 null lines of barley provides new information on the relationship of D hordein to malting performance

Six pairs of near isogenic lines, differing in the presence of absence of D hordein, were produced in backgrounds with different B and C hordein alleles by introducing a Hor 3 null gene discovered in a barley germplasm collection. Biochemical and cytochemical studies were performed on the harvested and malted grains. Statistical comparisons of the composition of the grains and their malting performance revealed that neither presence or absence of D hordein, nor gel protein levels related to differences in malting performance of the different lines. However, differences in hordein amount and in the grain ultrastructural characteristics could be related to some differences in malting performance between and within some pairs of lines.     

Susceptibility of wheat gluten to enzymatic hydrolysis following deamidation with acetic acid and sensory characteristics of the resultant hydrolysates

The effect of acetic acid and hydrochloric acid (HCl) deamidation pretreatment on the susceptibility of wheat gluten to enzymatic hydrolysis by Pancreatin and sensory characteristics of the resultant hydrolysates was investigated. At two degrees of deamidation (24% and 60%, with or without moisture-heating, respectively), wheat gluten pretreated by acetic acid deamidation was more susceptible to be hydrolyzed as evaluated by the hydrolysis degree, nitrogen solubility index, titratable acid amount and free carbohydrate content of the hydrolysates. Wheat gluten pretreated by acetic acid deamidation at a degree of 24% exhibited the highest susceptibility to enzymatic hydrolysis. Moisture-heating (121 °C, 10 min) in the deamidation pretreatment decreased the susceptibility of wheat gluten to enzymatic hydrolysis and the peptide factions of ≤3000 Da in the hydrolysates due to the formation of larger molecule weight aggregates. The hydrolysates prepared from acetic acid deamidated wheat gluten showed more intense glutamate-like and sauce-scented taste and better nutritional characteristics.     

普洱茶抗氧化特性的初步研究

本研究用不同的有机溶剂,分步萃取普洱茶的水提物,得到氯仿萃取物、乙酸乙酯萃取物、正丁醇萃取物和萃取后剩余物。采用Fe^2-H2O2-鲁米诺的化学发光法和有机自由基二苯基苦基偕腙肼（DPPH）的分光光度法,比较各提取物的清除自由基的能力。结果 提示普洱茶水提物中的乙酸乙酯萃取层组分和正丁醇萃取层组分对DPPH和羟自由基均有较强的清除能力。     

Influence of growth temperature on content, viscosity and relative molecular weight of water-soluble β-glucans in barley (Hordeum vulgare L.)

β-Glucan content and viscosity of water-soluble β-glucans have a considerable impact on the digestion of barley. Eight different 2-row barley (Hordeum vulgare L.) cultivars were grown in controlled environment chambers at five different temperatures until maturity. The samples were pearled, milled and analysed for their content of total, insoluble and soluble β-glucan. The water-soluble fraction was extracted at 37 °C, subjected to amylase treatment and freeze-dried. Water-soluble crude β-glucans were solubilised in distilled water, analysed for viscosity and their purity was checked by nuclear magnetic resonance. Molecular weights were profiled by size exclusion chromatography with RI detection. The content of total β-glucan varied from 4.0% to 7.4%, and was significantly affected by the growth temperature. An interaction between cultivar and growth temperature was observed for the total β-glucan content. The extractability was significantly affected by growth temperature, as there was recorded an increasing amount of water-soluble β-glucan with increasing growth temperature. Both the viscosity and the molecular weight of the water-soluble β-glucans increased with the growth temperature.     

Influence of sulphur application on hordein composition and malting quality of barley (Hordeum vulgare L.) in northern European growing conditions

The influence of sulphur (S) application on yield formation, hordein composition and malting quality of 2-rowed spring barley (Hordeum vulgare L.) was studied in Nordic conditions for the first time. In a greenhouse experiment, S deficiency was indicated when 10 mg S/kg soil or less was available, by an increase in the malate:sulphate ratio in leaves. The contents of aspartic acid and cystine in grains increased and decreased, respectively. Also a substantial decrease of total hordein and the proportion of B hordein, and an increase in the proportion of C hordein were associated with S deficiency. The effect of S was further assessed in field experiments by applying N and S in four different combinations prior to sowing. No apparent S deficiency occurred in the field conditions based on the malate:sulphate and grain N:S ratio. However, in a site where the grain S content was slightly but significantly increased by S application, a change in hordein composition and malting quality was observed. Thus even in S-sufficient conditions, the end use quality of malting barley may be affected by S application.     

Evaluation of thermomechanical pretreatment for enzymatic hydrolysis of pure microcrystalline cellulose and cellulose from Brewers’ spent grain

Microcrystalline cellulose (Avicel PH102) and Brewers’ spent grain (BSG) were subjected to Détente Instantanée Contrôlée (DIC) thermomechanical pre-treatment before exposure to cellulases (Celluclast 1.5 L). In a first part, we showed that the addition of β-glucosidase (Novozym-188) increased the hydrolysis yield of Avicel. A maximal theoretical yield (100%), was obtained for 5 and 10 g/L of Avicel using a mixture of Celluclast 1.5 L/Novozym-188. After DIC pre-treatments, the initial rate and final yield of hydrolysis decreased in comparison with those from untreated microcrystalline cellulose. This phenomenon may be due to the modification of the crystallinity of pure cellulose and the formation of inhibitors during the pre-treatment. In a second part, BSG was thermomechanically pre-treated and hydrolyzed. The results showed that the hydrolysis yield of BSG treated at pressure levels between 2 and 7 bar during 15 min was strongly improved compared to hydrolysis yield of untreated BSG. The optimized hydrolysis process, under intensive DIC conditions, achieved a glucose yield corresponding to 100% of the theoretical cellulose value. The morphology of BSG samples was studied with Scanning Electronic Microscopy (SEM) and highlighted that the structure of pre-treated BSG showed an important disruption compared to the rigid structure of untreated BSG.     

Hydrothermal and biotechnological treatments on nutraceutical content and antioxidant activity of rice bran

The effect of processing such as steaming, germination and parboiling on nutraceuticals and in vitro bioactive properties of rice bran from three different rice varieties namely Jyothi (pigmented), IR64 and Sona masuri (non-pigmented) were investigated. Within the varieties envisaged, pigmented Jyothi variety contained higher levels of vitamin E, soluble, bound and total polyphenol, flavanoids, free radical scavenging activity and total antioxidant activity. Direct steam exposure of bran resulted in an increase in, ether extractives and oryzanol, as well as retention of all the vitamin E components, bound polyphenols, flavonoids and decrease in soluble and total polyphenol content, free radical scavenging activity and total antioxidant activity compared to native. Parboiling as well as germination of paddy resulted in an increase, in the content of ether extractives and oryzanol, whereas other bioactive properties decreased compared to native. Hence it may be concluded that bioactive components and antioxidant properties were significantly higher in Jyothi bran compared to the other two paddy brans, and processing leads to changes in bioactive properties with maximum retention of bioactive components in the steamed bran.     

Influence of sprouting and elicitation on phenolic acids profile and antioxidant activity of wheat seedlings

Influence of germination and elicitation on nutraceutical potential of sprouted wheat was investigated. The seeds of three Polish winter wheat cultivars were elicited with 0.1% (w/v) of Saccacharomyces cerevisiae (SC) extract, 0.1% (v/v) Salix daphnoides (SD) bark extract and their combinations (SC/SD 1:1). Seeds were germinated for 4 day at 20 °C in darkness. For phenolic acids determination LC-ESI-MS/MS analysis was used. Antiradical ability, chelating power, reducing power and ability to prevent lipids against oxidation were determined. Bioaccessibility of antioxidative compounds was estimated using in vitro digestion method. Eleven phenolic acids (protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syryngic, p-coumaric, cis- and trans-ferulic, salicylic, trans- and cis-synapinic) were identified. Sprouting increased p-hydroxybenzoic, syryngic and p-coumaric acids level. The most effective elicitor for improving antioxidant activity of potentially bioaccessible phytochemicals was SD. Especially high effect was obtained for antiradical activity, chelating power and ability to lipid prevention.     

In vitro release of angiotensin-converting enzyme inhibitors, peroxyl-radical scavengers and antibacterial compounds by enzymatic hydrolysis of glycated gluten

The combined effect of gluten glycation and proteolysis on the release of compounds exhibiting in vitro angiotensin-converting enzyme (ACE) inhibitory, antioxidant and antibacterial activities was investigated. Model systems consisting of wheat gluten and glucose were heated at 120 °C for 45 min, 150 °C for 30 min and 220 °C for 30 min to produce various Maillard reaction products mimicking reactions occurring in bread crusts. Progress of the Maillard reaction was estimated through indirect measurement of Amadori compounds as 2-furoylmethyl-amino acids. Glycation was followed by digestion with Pronase E and ultrafiltration. The anti-hypertensive activity was measured as the ability to inhibit the activity of angiotensin I-converting enzyme involved in hypertension regulation. The Oxygen Radical Absorbance assay was used to measure the peroxyl radical scavenging capacity of the products and their effect on microbial growth of Escherichia coli ATCC 25922 and Staphlococcus aureus ATCC 25923 was also studied. Advanced products of the reaction enhanced the antioxidant and antibacterial properties of gluten hydrolysates and decreased the overall ACE inhibitory activity. Ultrafiltration provided a useful method for separating compounds (< 3000 Da) with ACE inhibitory activity and advanced Maillard reaction products (>3000 Da) which scavenged peroxyl radicals and inhibited the microbial growth.     

人工老化对大麦种子萌发过程中发芽特性及部分酶活性的影响

为了探究大麦种子的劣变机制,采用40℃与80%相对湿度的人工老化条件,对4个大麦品种进行老化处理,待种子风干后,测定其发芽特性、活性指标和5种抗氧化酶活性的变化。结果发现,大麦发芽特性指标在老化过程中呈反S型下降趋势;大麦种子浸出液相对电导率变化趋势不稳定,与发芽特性指标相关性不显著,四氮唑还原强度先上升后下降,过氧化氢酶活性先上升后下降,过氧化物酶、超氧化物歧化酶、谷胱甘肽还原酶、脱氢抗坏血酸还原酶活性总体呈下降趋势。种子发芽速率相关指标适合对大麦贮藏性的评价;种子浸出液相对电导率不能准确、灵敏地反映大麦种子的活性状况;四氮唑还原强度可作为部分大麦品种的活性评定指标;过氧化氢酶、过氧化物酶、谷胱甘肽还原酶活性与大麦种子老化过程密切相关。     

Effects of sulphur on yield and malting quality of barley

Eight field experiments were conducted at four sites in the UK in 2003 and 2004 to investigate the effects of sulphur (S) application on yield and malting quality of barley. Significant yield responses to S additions were obtained in five out of the eight experiments, with yield increases ranging from 0.2 to 1.2 t/ha (4.7–22.5%). At the two most S-deficient sites, S application significantly increased malt diastatic power, alpha-amylase activity, friability and homogeneity, and decreased (1→3,1→4)-β-glucan concentration in the wort, indicating an improved endosperm modification during malting. Sulphur applications also significantly increased the concentration of S-methylmethionine (the precursor of dimethylsulphide) in kilned malt, which could impact on beer flavour. When the supply of N was limiting, S applications decreased grain N concentration due to a dilution effect as a result of increased grain yield. In some cases, S applications resulted in decreased grain size. At sites non-deficient or marginally deficient in S, applications of S had little effect on grain or malting quality parameters. The need to maintain an adequate S supply to barley for both yield and malting quality was demonstrated.