Induction of cellulases and hemicellulases from Neurospora crassa under solid-state cultivation for bioconversion of sorghum bagasse into ethanol

The cellulolytic and hemicellulolytic system from the mesophilic fungus Neurospora crassa was produced under solid-state cultivation (SSC) on wheat straw and wheat bran mixtures. Following optimization of nitrogen source, pH and initial moisture of the growth medium, yields as high as 492.8, 1.08, 26.7, 297.8 and 0.132 (in U g^?1 of carbon source) were obtained for endoglucanase, exoglucanase, β-glucosidase, xylanase and β-xylosidase, respectively. The potential of the multienzyme system was demonstrated for hydrolysis of sorghum bagasse (SB) into fermentable carbohydrates. N. crassa cells were found able to assimilate the majority of the released sugars and generated limited levels of other metabolic products during simultaneous saccharification and fermentation of this valuable substrate into ethanol.     

Value-addition of agricultural wastes for augmented cellulase and xylanase production through solid-state tray fermentation employing mixed-culture of fungi

Solid-state fermentation (SSF) was performed to evaluate the potential of agricultural residues for the production of cellulase and hemicellulase using individual and mixed cultures of Aspergillus niger and Trichoderma reseei. The maximum filter paper (FP) cellulase activity of 13.57 IU/gram dry substrate (gds), 22.89 IU/gds and 24.17 IU/gds and β-glucosidase activities of 21.69 IU/gds, 13.58 IU/gds and 24.54 IU/gds were obtained with wheat bran medium at 96 h incubation period with A. niger, T. reseei and mixed-cultures of A. niger and T. reseei, respectively. Mixed-culture SSF using rice straw supplemented with wheat bran in the ratio 3:2 resulted in higher FP cellulase, β-glucosidase, endoglucanase (CMCase) and xylanase activities, compared to the activities obtained using mono-cultures. Similarly, higher FP cellulase, β-glucosidase, CMCase and xylanase activities of 35.8 IU/gds (96 h), 33.71 IU/gds (96 h), 131.34 IU/gds (120 h) and 3106.34 IU/gds (120 h) were achieved in the tray fermentation using rice straw with wheat bran in the ratio of 3:2. Results of present investigation showed that higher cellulase activity and an optimal combination of cellulase and β-glucosidase can be achieved through mixed-culture SSF in trays. The approach of utilizing negative cost agricultural wastes through tray fermentation for cellulase and hemicellulase production is expected to serve the objectives of: (a) management of wastes which would otherwise cause environmental pollution problems; (b) production of hydrolytic enzymes at low cost and; (c) simple technique requiring no sophisticated instruments with practical applications.     

Alkaline Hydrogen Peroxide Extraction of Wheat Bran Non-starch Polysaccharides

An extraction method based on the use of dilute alkaline solutions of hydrogen peroxide removing the lignin present in lignocellulosic materials (Gould, J. M. Biotechnology and Bioengineering 26 (1984) 46–52), was studied for the extraction of non-starch polysaccharides (NSP) from wheat bran. The influence of extraction temperature and time, hydrogen peroxide (H₂O₂) concentration and percentage of bran in the alkaline hydrogen peroxide solution on the extraction yields of non-cellulosic glucose, arabinose and xylose residues were investigated. Extraction for 4 h at 60 °C with an alkaline (pH 11.5) H₂O₂-solution (2·0%) and 2·0% wheat bran in the extractant yielded an alkaline extract containing 77% of the wheat bran total arabinose content, 65% of the total xylose content and 86% of the total non-cellulosic glucose content.     

Modeling the effects of solid state fermentation operating conditions on endoglucanase production using an instrumented bioreactor

Cellulase production is one of the most critical steps in the economics of second generation ethanol. Although solid-state fermentation (SSF) is an attractive process for the production of enzymes, SSF is highly limited by the difficulty in controlling the operating variables which affect microbial growth and metabolites production. In this context, this work evaluates the effects of operational conditions on endoglucanase production by a selected strain of Aspergillus niger cultivated under SSF using an instrumented lab-scale bioreactor equipped with an on-line automated monitoring and control system. The effects of air flow rate, inlet air relative humidity and substrate initial moisture on endoglucanase production were evaluated using a statistical design methodology. A correlation coefficient of 0.9106 and a calculated value of F, 5.46 folds higher than the listed value (P-value < 0.05) allowed the modeling of endoglucanase production under different process conditions. Higher endoglucanase production (56.1 U/g) was achieved for a selected condition of substrate initial moisture of 72%, air inlet humidity of 70%, and flow rate of 20 mL/min. A significant increase in endoglucanase production was also found to be achieved under forced aeration conditions (50.2 IU/g) compared to static conditions (29.8 IU/g) after 72 h of cultivation. Besides, respirometric analysis revealed that the total amount of CO₂ produced was linearly correlated with enzyme production (R² of 0.988). The bioreactor system used, as well as the methodology employed herein, was very effective in evaluating the influence of operational variables on enzymes production under SSF.     

Estimated Glycemic Index and Dietary Fiber Content of Cookies Elaborated with Extruded Wheat Bran

The increasing demand for high-fiber products has favored the design of numerous bakery products rich in fiber such as bread, cookies, and cakes. The objective of this study was to evaluate the dietary fiber and estimated glycemic index of cookies containing extruded wheat bran. Wheat bran was subjected to extrusion process under three temperature profiles: TP1;(60, 75, 85 and 100 °C), TP2;(60, 80, 100 and 120 °C), and TP3;(60, 80, 110 and 140 °C) and three moisture contents: (15, 23, and 31 %). Cookies were elaborated using extruded wheat bran (30 %), separated into two fractions (coarse and fine). The dietary fiber content of cookies elaborated with extruded wheat bran was higher than the controls; C0 (100 % wheat flour) and C1 (30 % of no extruded bran coarse fraction) and C2 (30 % of no extruded bran fine fraction). The higher values of dietary fiber were observed on cookies from treatments 5 (TP1, 31 % moisture content and coarse fraction) and 11 (TP2, 31 % moisture content and coarse fraction). The estimated glycemic index of cookies ranged from 68.54 to 80.16. The dietary fiber content of cookies was increased and the lowest glycemic index corresponded to the cookies elaborated with extruded wheat bran. Cookie made with the treatment 11 had a better dietary fiber content and lower estimated glycemic index.     

Value added waste of Jatropha curcas residue: Optimization of protease production in solid state fermentation by Taguchi DOE methodology

The oil extraction of Jatropha curcas created the large amount of the by-product from its seeds. An application of solid-state fermentation (SSF) was considered to be of value to these raw materials. This study investigated the potential of a utilization of deoiled J. curcas seed cake as substrate for protease productions by Aspergillus oryzae. While various parameters for SSF was conventionally individually optimized, five parameters were simultaneously examined based on Taguchi method. The effect of three different levels of five factors, including moisture content of substrate, inoculums size, incubation temperature, type of porous substrate and incubation time were examined. The optimum conditions for the protease production by A. oryzae obtained from this experiment were 45% moisture content of substrate, 10% inoculums size, 30 °C incubation temperature, deoiled J. curcas seed cake mixed with cassava bagasse ratio 4:1 as porous substrate at 84 h of incubation time. By adjusting the conditions to these optimum levels, the protease production increased up to 4.6 times as many as the protease yield from the non-optimizing experiment. The use of statistical approach, Taguchi method, provided a satisfactory outcome in defining the optimum conditions for protease production by A. oryzae. Further, the utilization of deoiled J. curcas seed cake as substrate for SSF was proven as the suitable practice for this agricultural waste, in order to develop for an industrial use.     

木聚糖酶高产菌株的筛选及产酶条件

从本实验室保存的46株纤维素分解菌中筛选出1株可高效降解半纤维素的木聚糖酶高产菌株APS35,其酶活力高达31.801IU.g-1,比APS02（对照）高4.91倍。产酶条件优化结果表明,以水稻秸秆为底物,APS35产木聚糖酶的最适条件：培养温度28℃,培养时间3-4d,稻草粉与麸皮比4∶1,接种量10%,氮源为酵母膏（总氮量0.4%）,pH为4.0,吐温80浓度0.4%。在此条件下的木聚糖酶活力为32.024IU.g-1,与优化前无显著差异。     

Changes in bran structure by bioprocessing with enzymes and yeast modifies the in vitro digestibility and fermentability of bran protein and dietary fibre complex

Bran is a good source of dietary fibre, phytochemicals, and also protein, but highly insoluble and recalcitrant structure of bran hinders accessibility of these components for gastrointestinal digestion. In the present work, influence of bioprocessing on the microstructure and chemical properties of rye bran and wheat bread fortified with the rye bran were studied. In vitro protein digestibility, and release of short chain fatty acids (SCFA) and ferulic acid in a gut model were studied. Bioprocessing of rye bran was performed with subsequent treatments with cell-wall hydrolysing enzymes (40 °C, 4 h) and yeast fermentation (20 °C, 20 h). Bioprocessing of rye bran resulted in reduced total dietary fibre content, caused mainly by degradation of fructan and β-glucan, and increased soluble fibre content, caused mainly by solubilisation of arabinoxylans. Microscopic analysis revealed degradation of aleurone cell wall structure of the bioprocessed rye bran. Bioprocessing caused release of protein from aleurone cells, assessed as a larger content of soluble protein in bran and a higher hydrolysis rate in vitro. Bioprocessed bran had also faster SCFA formation and ferulic acid release in the colon fermentation in vitro as compared to native bran.     

添加酶制剂对饲用苎麻与麦麸混合青贮品质的影响

为了探究酶制剂对苎麻混合青贮品质的影响,本试验在苎麻与麦麸(80%苎麻+18%麦麸+2%蔗糖)混合青贮中分别添加10 mL蒸馏水(对照组)、纤维素酶2 g/kg、纤维素酶2 g/kg+半纤维素酶0.4 g/kg、纤维素酶2 g/kg+半纤维素酶0.4 g/kg+果胶酶0.7 g/kg,30天后开包取样分析各项指标。结果表明,苎麻与麦麸混合青贮发酵品质和营养品质均较好。与对照相比,添加酶制剂显著提高了碳水化合物、干物质和粗蛋白含量(P<0.05),显著降低了中性洗涤纤维、酸性洗涤纤维、粗纤维的含量和氨态氮/总氮比值(P<0.05),但是对pH值、乳酸、乙酸和丙酸含量无显著影响(P<0.05)。本试验条件下,添加酶制剂能够有效提高苎麻混合青贮的品质,单独添加纤维素酶与复合酶制剂无显著性差异,从节约成本角度考虑,单独添加纤维素酶组为最佳选择。     

Wet grinding and microfluidization of wheat bran preparations: Improvement of dispersion stability by structural disintegration

The enrichment of liquid food matrix with wheat bran has not yet been explored. This study investigated the impact of disintegrating wheat bran preparations on their stability at high moisture content. Three wheat bran preparations – standard bran, peeled bran and aleurone rich fraction – were modified by dry grinding, enzymatic degradation, wet grinding and microfluidization. The sedimentation of processed preparations was evaluated in water solution and related to their physical structure, solubilized compounds and suspension viscosity. In dry ground preparations mixed in water (5% w/w), most of the particles sedimented already in 5 min. Wet grinding disintegrated the physical structure of bran preparations (d₅₀ = 10–16 μm), causing improvement of particle stability due to reduction of gravitational sedimentation. Enzymatic treatment with xylanase efficiently increased the total solubility of the bran preparations (from 18–24% to 40–50%), but the higher solubility was not related to the better stability of particles. Microfluidization of peeled bran and aleurone increased the viscosity and stability of dispersions. The higher viscosity of the microfluidized dispersions was likely correlated with the better homogenisation of the particles, and also with the modified microstructure of treated bran preparations. Disintegrated wheat bran preparations showed high potential for beverage applications.     

Moisture Distribution in a Mature Soft Wheat Grain by Three-Dimensional Magnetic Resonance Imaging

Moisture-tempering is typically performed prior to first break in wheat milling operations to enhance separation of bran, germ, and endosperm. To improve the milling performance and to increase yield, a better knowledge of moisture distribution and migration in individual wheat grains during tempering is essential. The research described herein was conducted to demonstrate the non-destructive measurement of the three-dimensional (3D) distribution of moisture in a single wheat grain. A 3D magnetic resonance imaging (MRI) technique was adapted for the probing of single wheat grains at storage moistures (c.12% wet basis). The technique is demonstrated in this report on one grain. A 3D projection reconstruction (3DPR) technique was used to acquire high resolution proton density images. The spatial resolution of the images was 94×94×94 μm³. The 3D proton density images were related to the 3D moisture distribution in the wheat grain. At 12% moisture content, the moisture distribution in the starchy endosperm of the wheat was not uniform. The variation of the moisture distribution was from 7·3% to 16·4% wb.     

Dephytinization of wheat bran by fermentation with bakers' yeast,incubation with barley malt flour and autoclaving at different pH levels

Wheat bran is an important source of dietary fiber but also contains considerable amounts of phytic acid, which is known to impair mineral absorption. The present study was conducted to investigate the phytic acid reduction in coarse and fine wheat bran by fermentation with the different levels of bakers' yeast (3, 6 and 9%) for 8 h at 30 °C, incubation with the different levels of barley malt flour (2.5, 5.0, 7.5 and 10.0%) for 8 h at pH 5.2 and 55 °C, and autoclaving at the different pH levels (pH 5.0, 4.5, 4.0 and 3.5) adjusted with acetic acid for 2 h. The phytic acid content of the wheat bran was effectively reduced by all treatments, and the phytic acid lost was in the range of 88.4–96.9%. Without addition of yeast or malt flour, or autoclaving without pH adjustment, the phytic acid content of the bran samples was reduced at most to 44.9% of the initial amounts under the investigated conditions. Increasing the concentration of yeast or malt flour or decreasing the pH towards 3.5 did not enhance the phytic acid reduction. The most reduction occurred after 2 h of yeast fermentation and malt flour incubation, and after 30 min of autoclaving, which made up 92–98% of the total phytic acid loss. Extending the treatment periods contributed nominally to further increase in the phytic acid reduction, and the rate of the phytic acid loss decreased progressively.     

秸秆带状覆盖对旱地冬小麦产量及土壤水分的影响

为探讨西北半干旱雨养条件下秸秆带状覆盖麦田水分利用特征和增产效果,通过田间试验,以露地条播为对照(CK),研究了3种不同覆盖处理[秸秆带状覆盖常规条播(SM1)、秸秆带状覆盖宽幅条播(SM2)和全膜覆土穴播(PM)]对旱地冬小麦田土壤含水量、小麦产量和水分利用效率的影响。结果表明,秸秆带状覆盖和全膜覆土穴播均可显著改善小麦全生育期0~20cm以及开花前20~90cm土壤墒情,但开花后20~90cm以及全生育期90~200cm土壤墒情普遍不如CK。3种覆盖处理均能促进冬小麦对土壤贮水的利用,显著提高开花至成熟阶段的耗水量及其占总耗水量的比例。秸秆带状覆盖和全膜覆土穴播生育期耗水量分别比CK增加4.6%和7.6%。秸秆带状覆盖在孕穗前0~200cm土壤墒情与全膜覆土穴播无显著差异,孕穗期开始则好于全膜覆土穴播;全膜覆土穴播0~200cm土壤贮水消耗量显著高于秸秆带状覆盖,而开花至成熟阶段的耗水量及其占总耗水量的比例则略低于秸秆带状覆盖。3种覆盖处理均显著提高产量和水分利用效率,PM、SM1和SM2较CK分别增产36.8%、29.7%和27.5%,水分利用效率分别提高27.3%、23.9%和22.7%。产量与生育期耗水量呈显著正相关(r=0.97*)。覆盖处理中,全膜覆土穴播产量虽最高,但从产量、水分利用效率和经济效益角度综合考虑,秸秆带状覆盖优于全膜覆土穴播。因此认为,秸秆带状覆盖是一种更加高产高效、适宜在西北半干旱雨养区推广的覆盖种植方式。     

Effect of solid state fermentation by Enterococcus faecalis M2 on antioxidant and nutritional properties of wheat bran

Wheat bran is the main by-product during wheat flour processing. Although wheat bran is rich in the bioactive compounds and antioxidant capacity, it is not widely utilized in its natural state. To improve the antioxidant and nutritional properties of wheat bran, a dominant strain, Enterococcus faecalis M2, was screened from lactic acid bacteria (LAB) isolated from various foods. Following solid state fermentation, the soluble dietary fiber content of wheat bran nearly quadrupled compared to the raw material. Total proportion of phenols, flavonoids, alkylresorcinols, along with the antioxidant capacity and free radical scavenging rate were significantly improved, particularly the ferulic acid content increased by 5.5 times. Additionally, the free amino acid content increased with degradation of wheat bran protein, whereas the level of anti-nutrient phytic acid decreased. The results of this study could provide an effective method for biological modification of wheat bran, which further enhance the health benefit and utilization of bran.     

Strawboard from vapor phase acetylation of wheat straw☆

Commercial ground wheat straw was used in a central composite response surface experimental design to examine four acetylating process variables: reaction temperature, reaction time, initial moisture content of straw, and the vapor flow rate of chemical reagent. The response variable was acetyl content determined as a function of straw weight gain. Diphenylmethyane diisocyante was used as a binder to prepare board samples with a hot press. Equilibrium moisture content (EMC) was determined at 65 and 90% RH at 27°C, and dimensional stability was determined using a humidity cycle of 30–90% RH at 27°C. ASTM D1037-93 standard method for a 3-point flex test was used to measure mechanical properties. The microstructures of both treated and untreated wheat straw and boards were observed with a scanning electron microscope. The vapor phase acetylation system used acetylated ground wheat straw to a 24% weight gain (dry weight basis). A mathematical model (R²=0.97) was developed to predict the weight gain as a function of the four acetylation processing variables. The maximum reduction in all strawboard properties occurred at the highest weight gain (24%). The strawboard EMC decreased (30% maximum reduction) as weight gain increased at both 65 and 90% RH. The strawboard dimensional stability increased as the weight gain increased (maximum reductions of 80% in thickness swell and 50% in linear expansion). The initial mechanical properties of the strawboards decreased as the weight gain increased (maximum reductions of 64% in strength and 48% in stiffness). The density of the strawboards decreased as the weight gain increased (23% maximum reduction). SEM micrographs showed no physical evidence of structural damage to cell walls from the acetylation.     

淡紫拟青霉E7菌株固体放大发酵产孢培养基和工艺条件研究

为探讨淡紫拟青霉E7菌株固体放大发酵因子间的关系以获得大量孢子用于田间试验,通过正交试验设计优化固体发酵培养基成分和培养条件组合,并测定外加碳源、氮源、无机盐和装料方式对产孢量的影响。结果表明：以玉米粉+甘蔗渣+麸皮+壳聚糖组成的正交2号配方为最佳复合培养基质,分生孢子产量达7.13×109个/g,以发酵液pH+液相发酵终点+温度+初始含水量组成的正交4号配方为优适培养条件,分生孢子产量达8.97×109个/g;该菌株在优化后的培养基配方中添加0.4％蔗糖、0.2％蛋白胨、0.002％硫酸锰,以双层纱布袋装料按优化后的培养条件放大培养,产孢量最高可达到8.22×1010个/g。优化后的E7菌株发酵产孢培养基基质用量少,发酵成本低,适合于固体放大发酵生产淡紫拟青霉孢子。     

Production of gluten-free wheat starch by peptidase treatment

The potential of peptidase-containing bran extracts from germinated cereals (wheat, emmer, barley) and a peptidase preparation from Aspergillus niger (AN-PEP) to degrade gluten in wheat starch below the threshold for gluten-free foods of 20 mg/kg was compared. The gluten-specific peptidase activity of the peptidases was determined by using gliadin as a protein-based substrate as well as the two celiac-active peptides PQPQLPYPQPQLPY (α-gliadin) and SQQQFPQPQQPFPQQP (γ-hordein). The peptidase activity of AN-PEP exceeded the activities of bran from germinated cereals by a factor up to 690,000. Three wheat starches with initial gluten contents of 110, 1679, and 2070 mg/kg, respectively, were incubated with bran extracts and AN-PEP, lyophilized, and residual gluten was quantitated by a competitive ELISA. Unlike peptidases from bran extracts, AN-PEP was capable of degrading gluten below 20 mg/kg in all starches. The absence of gluten in AN-PEP-treated starches was confirmed by liquid-chromatography-mass spectrometry. The properties of gluten-free starches were comparable to the native starches with the exception of a reduced viscosity after AN-PEP treatment. This problem could be overcome by using higher enzyme concentrations and shorter incubation times or by optimizing AN-PEP production for lower residual α-amylase activity.     

Biochemical and sensory evaluation of wheat bran supplemented sorghum bread

Addition of wheat bran to sorghum flour (Dabar cultivar) at two extraction rates 72% and 80% resulted in lowering reducing sugars. The percent decreasewas 75.6% compared with the control at the end of fermentation period. There was a highly significant (p 0.05) increase in crude fiber content as a result of addition of wheat bran. The increase was from 0.8 to 5.2 and from 0.5 to 5.3% for the 80% S/WB and 72% S/WB blends, respectively. Sorghum bread containing wheat bran was lower in reducing sugars and showed a significant increase (p 0.05) in starch content. Sorghum bread containing wheat bran resulted in a lower in vitro protein and starch digestibilities.     

自然降水条件下耕作方式对小麦生长、产量及土壤水分的影响

为探寻小麦高产节水耕作技术,在2011-2013小麦生长季,通过定位试验,以安徽大面积主推小麦品种济麦22为材料,比较分析了在自然降水条件下秸秆还田+深耕+播后镇压、深松+播后镇压、旋耕+播后镇压、旋耕和复式播种作业5种耕作方式下淮北地区小麦生长发育、产量及土壤水分的差异。结果表明,各耕作处理下小麦田间出苗率均在70%以上,基本可以满足小麦生产对田间出苗的要求。在5个处理中,旋耕+播后镇压处理出苗率最高,达83%;复式播种作业处理的单株次生根数最高,2年在生育后期分别达到24.2和28.9条;旋耕+播后镇压处理2年平均及各年度的籽粒产量均最高,分别为8 206.8、8 175.5和8 238.0kg·hm-2,复式播种作业次之,秸秆还田+深耕+播后镇压最低,处理间差异显著。深松和旋耕处理在小麦生育后期0~20cm土层土壤水分含量较低,而40~60cm土层土壤水分含量较高。本试验条件下,旋耕+播后镇压是小麦兼顾高产节水的最优耕作播种方式。     

Effects of region and cultivar on alkylresorcinols content and composition in wheat bran and their antioxidant activity

This study evaluated the effects of cultivar and region on the composition of alkylresorcinols (ARs) of 24 wheat bran samples from 6 cultivars grown in four locations (Bath, Nairn, Palmerstone and Ridgetown) in Ontario (ON), Canada, using gas chromatography/mass spectrometry (GC/MS). Total phenolic content (TPC) of wheat bran extracts was determined by the Folin–Ciocalteau method and the antioxidant activity of wheat bran extracts was measured by 2, 2-Dipheny-1-picryhydrazyl radical (DPPH) scavenging activity and oxygen radical absorbance capacity (ORAC). The highest ARs content (μg/g) was found in cultivars Emmit (1522), Harvard (1305), Warthog (1170), and Superior (853), grown in Ridgetown. The relative saturated and unsaturated ARs (%) were 89 and 11, respectively. Total ARs content, their composition, TPC and antioxidant activity of wheat bran extracts were significantly (P < 0.05) affected by location and cultivar and their interaction. TPC, %DPPH_dis, and ORAC values for different wheat bran sample extracts ranged from 3 to 58 (mg FAE/g), 5 to 68 (%), and 6 to 94 (μmol TE/g), respectively. Our work provides a detailed examination of region and cultivar effects on potential of ARs in wheat bran and the results can be used for screening and breeding purposes.