毕赤酵母液态发酵产木聚糖酶条件研究

用麦麸、米糠农业废弃物作为主要原料生产木聚糖酶,对毕赤酵母液态发酵产木聚糖酶的条件进行研究。结果表明,最适产酶条件:以质量浓度20%的麦麸作为碳源,4%酵母浸膏作为氮源,调节初始pH值5.5,培养温度30℃,摇瓶发酵装液量6%,培养时间130 h,在此条件下毕赤酵母液态发酵产木聚糖酶最高活力达到2192 IU/mL。     

Development of a solid-state fermentation process for production of an alpha amylase with potentially interesting properties

Ca-independency with potential activity and stability at low pH are among the most interesting characteristics of α-amylase in starch industry. In this attempt the synergetic effect of low pH on activity of crude Ca-independent α-amylase isolated from a native Bacillus sp. KR-8104 in solid-state fermentation (SSF) was studied using wheat bran (WB) as a substrate. The effects of different parameters including moisturizing agents, solid substrate to moisture ratio, particle size, incubation temperature and period, inoculum (v/w) and supplementation with 1% (w/w) different carbon and nitrogen sources on enzyme production were investigated. Maximum enzyme production of 140 U/g dry fermented substrate was obtained from wheat bran moistened with tap water at a ratio of 1:1.5 and supplemented with 1% (w/w) NH₄NO₃ and 1% (w/w) lactose after 48 h incubation at 37 °C. Even though the production of α-amylase was lower at 40 and 45 °C, the viable cell count was higher. In addition response surface methodology (RSM) was applied to find optimum conditions of temperature and pH on crude amylase activity. Using central composite design (CCD) a quadratic mathematical model equation was derived for the prediction of enzyme activity. The results showed that the model was in good agreement with experimental results, with R² = 0.90 (p < 0.0001) and the low pH has a synergetic effect on enzyme activity at higher temperature.     

Rennin-like milk coagulant enzyme produced by a local isolate of Mucor

Among 20 isolates of Mucor isolated from various environments in Jordan and found to produce a rennin-like acid protease, known as Mucor rennin-like enzyme (MRE), Mucor J20 was found to produce the highest level of MRE. The optimum incubation conditions for enzyme production in a fortified wheat bran mixture using solid-state fermentation were 3–4 days at 30°C. The highest MRE activity (185–200 rennin units or RU) was produced in a medium containing wheat bran and lentil straw (1 : 1 w/w) moistened with whey, and incubated in clay pots at 30°C for 4 days. A slightly lower activity value (178 RU) was found when using a mineral salt solution or distilled water instead of whey, or when using wheat bran alone with whey. At pH 4, the MRE retained its complete activity (100%) for 6 weeks at 5°C and 10°C, and for 3 and 2 weeks at 20°C and 30°C, respectively. After heating at 60°C for 10 min, the enzyme lost its activity at all pH levels used (pH 2–8). The crude extract of MRE was successfully applied in the manufacture of a cheese curd.     

Optimization of the fermentation medium for alpha-galactosidase production from Aspergillus foetidus ZU-G1 using response surface methodology

ABSTRACT:  The optimization of fermentation medium for alpha-galactosidase production by Aspergillus foetidus ZU-G1 was investigated in shaker flask fermentation. A one-factor-at-a-time experiment was used to screen the preferable nutriment (carbon sources, nitrogen sources, and essential elements) for alpha-galactosidase production. A fractional factorial design was used to screen the main 5 factors, soybean meal, wheat bran, KH₂PO₄, FeSO₄·7H₂O, and the medium initial pH, that affected the production of alpha-galactosidase. The central composite experimental design was further adopted to derive a statistical model for optimizing the composition of the fermentation medium. The experimental results showed that the optimum fermentation medium for alpha-galactosidase production by Aspergillus foetidus ZU-G1 was composed of 3.2% soybean meal (w/v), 2% wheat bran (w/v), 0.1% KH₂PO₄ (w/v), and 0.05% FeSO₄·7H₂O (w/v); initial medium pH was 6.31. The results further predicted that alpha-galactosidase activity reached 64.75 U/mL after 96-h incubation in this medium, which was approximately 7 times higher than that incubated in the nonoptimized medium. The time course of alpha-galactosidase production in the optimized medium composition was also carried out to validate the model.     

Single‐cell protein production through microbial conversion of lignocellulosic residue (wheat bran) for animal feed

Agricultural residue (wheat bran) rich in carbohydrates was utilized in the fermentation process to produce microbial biomass. Single‐cell biomass consists of the dried cells of microorganisms, which are used as protein supplements in human food and in animal feed. In the present study, two different microorganisms (Candida utilis and Rhizopus oligosporus) were studied for biomass production. To enhance the nutritional contents of wheat bran, a number of different fermentation parameters (effect of inoculum size, age of inoculum, incubation period, moisture to substrate ratio and incubation temperature) were optimized. Maximum yield was obtained at an inoculum size of 10% (v/w), with the age of the inoculum being a 48 h old culture. A fermentation period of 48 h was found to give the maximum protein yield and viable counts of yeast cells and mould hyphae. The microorganisms showed good growth at 30 °C. After complete optimization of the fermentation parameters, a batch of wheat bran was fermented with C. utilis and R. oligosporus under the optimized conditions, resulting in a maximum crude protein yield of 41.02% compared with the 4.21% crude protein of the non‐fermented wheat bran. Copyright © 2015 The Institute of Brewing & Distilling     

金针菇FV_(8812)菌株深层发酵工艺的研究

金针菇FV_8812(Flammnlina Velutipes)菌株是我们从国内外收集到的25株菌株中,经筛选获得的一株适合深层发酵的高产优质菌株。本文报道了该菌株的适宜发酵培养基、摇瓶发酵条件及50升发酵罐的扩大试验结果。结果表明,其适宜发酵培养基的组成为5.0％玉米粉、3.0麸皮、0.1％KH_2PO_4、0.05％MgSO_4·7H_2O、10μg/100ml VB_1、50μg/100ml VB_2;适宜的摇瓶发酵条件为:培养温度20～25℃培养基起始pH6.0～7.0,摇瓶装量500ml,装100～160ml,种子培养时间3～4天,接种量10～15％,摇瓶转速120～200rpm添加消泡油不少于0.1％为宜;经50升发酵罐扩大试验,菌体干收率3.94％(W/V),达到了工厂化发酵生产食用菌菌丝体的要求,因而具有工业投产意义。     

耐盐米曲霉制曲及复合酶分泌条件的研究

该文研究了耐盐米曲霉制曲的产酶特性,并且通过对耐盐米曲霉在不同作用条件下制得的曲料的蛋白酶活力和淀粉酶活力的变化分析,探讨了不同原料配比、水分添加量、制曲温度和蒸料时间等因素对耐盐米曲霉制曲效果的影响,为米曲霉在工业生产中的研究和应用提供理论依据.研究表明,耐盐米曲霉制曲产生的中性蛋白酶和碱性蛋白酶的酶活较高,酸性蛋白酶和淀粉酶酶活较低.复合酶分泌的最佳工艺条件为:原料配比为豆粕:麸皮=2:3,加水量为120％,蒸料时间为30min,30℃制曲42h,此条件下酸性蛋白酶和淀粉酶活力有明显的提高.     

米糠和麦麸膳食纤维的制备研究

探讨了米糠半纤维素和麦麸膳食纤维的提取工艺。结果表明 :( 1 )料液比 1∶1 0 ,6 0℃浸提 3h ,米糠水溶性半纤维素提取率 1 38% ;( 2 )料液比 1∶1 0 ,0 5mol/L的NaOH 2 5℃浸提 3h ,米糠碱溶性半纤维素提取率 8 86 % ;( 3) 0 6 %的NaOH ,α 淀粉酶加入量 0 4% ,70℃浸提 1 5h ,麦麸膳食纤维提取率达 6 6 2 7%。     

黑曲霉耐酸性α-淀粉酶的固态发酵条件研究

研究了固态发酵条件对黑曲霉(Aspergillus niger)PZ301合成酸性α-淀粉酶的影响。结果表明,固态发酵最佳培养基组成:麸皮7.0g,葡萄糖0.07g,淀粉0.21g,(NH4)2SO40.07g,起始pH 4.2;最佳培养条件为培养温度35℃,料水比为1∶1,接种量3mL(孢子浓度为107个/mL)。在上述条件下培养72h,酶活力可达19.6IU/g干培养基。另外还测定了PZ301固态发酵中生物量的变化,PZ301菌株的生物量和耐酸性α-淀粉酶在72h时均达到最高,这表明耐酸性α-淀粉酶系同步合成型酶。     

Milk‐clotting enzymes produced by Aspergillus flavo furcatis strains on Amazonic fruit waste

Six strains of Aspergillus flavo furcatis were screened to investigate milk‐clotting enzyme production by fermentation in natural liquid medium. The growth media comprised extracts of cupuaçu exocarp+rice bran [10% or 20% (v/v) CE+RB] and açai waste+rice bran [10% or 20% (v/v) AW+RB] with or without supplementation of 0.1% (w/v) yeast extract and 0.5% (w/v) gelatin. Significant values of milk‐clotting activity were determined by A. flavo furcatis DPUA 1461 and DPUA 1608, in the standard and natural media, respectively. According to criteria of clot and whey formations, 8.3% of the samples tested were classified as strong coagulation, 41.70% showed weak coagulation and in 50% was not observed milk coagulation. The enzyme optimal action of DPUA 1608, the selected strain, was at 40 °C and pH 7.0. Milk‐clotting proteases were inhibited by pepstatin (94.72%) and moderately inhibited by the others metal ions tested.     

PS0312 菌株发酵产淀粉酶及温度与pH 值对酶活力的影响

PS0312 菌株是具有特殊生物学特性的青霉菌株。以三角瓶固体培养研究不同条件与菌株产淀粉酶的关系,以及温度与pH 值对酶活力的影响。结果表明：PS0312 菌株以麸皮30.04%、大豆饼粉3.70%、谷壳3.70% 及55.56% 蒸馏水组成的培养基固体发酵湿麸曲淀粉酶产量最高。单因子试验结果表明：以培养基pH3、108 个/mL的种子液接种量1.85%、培养温度28℃、培养时间96h 产淀粉酶量最大。PS0312 菌株产淀粉酶在pH2.0～10.0 内具较高活性,酶活大小与pH 值的关系成双峰形;pH3 的条件下酶活性最高,相对酶活100%;pH9 的条件下酶活次之,相对酶活84.98%。酶反应最适温度为60℃,90℃条件下相对酶活为39.06%。研究表明：PS0312 菌株发酵产淀粉酶是一种在强酸、强碱条件下都具有较高的酶活性和耐高温的特殊的酸性、中温淀粉酶,可在较宽的温度与pH 值范围下应用。     

好食脉孢霉发酵麦麸制备可溶性膳食纤维及其理化性质

采用好食脉孢霉对小麦麸皮进行固态发酵制备可溶性膳食纤维（Soluble dietary fiber,SDF）,通过单因素结合响应面法Box-Behnken探究发酵过程中含水量、接种量、发酵温度、发酵时间对可溶性膳食纤维得率的影响,确定培养基的最佳发酵条件。同时对发酵过程中纤维素酶活性和木聚糖酶活性进行测定,并研究发酵前后SDF的理化性质。结果表明:当发酵温度为29℃、接种量为11%（v/w）、含水量为74%（v/w）、发酵时间为83.5 h时SDF得率最高,为13.41%,比发酵前提高了1.05倍。发酵过程中纤维素酶活性与木聚糖酶活性均与SDF得率呈正相关。发酵后SDF溶解性、吸附葡萄糖能力、吸附胆固醇能力（pH=2和pH=7）和DPPH清除能力比发酵前分别提高了1.14、1.76、5.36、4.61和1.62倍,为麦麸SDF作为食品添加剂提供理论基础。     

黑曲霉果胶裂解酶和聚半乳糖醛酸酶合成的发酵调控

通过对黑曲霉(Aspergillus niger)wz 003液态发酵过程的调控,制备了高活性的果胶裂解酶,并讨论了发酵过程中聚半乳糖醛酸酶活力的变化情况。实验表明,发酵产酶培养基组成为(g/L):麸皮60,玉米粉40,酵母膏20,胡萝卜12,CaCO_3 10。培养条件为:初始pH 6.5,30℃,接种量8%,培养2 d。此时果胶裂解酶活力为375.3 U/mL,为基础条件下的6.225倍,而聚半乳糖醛酸酶的合成得到了有效控制,酶活仅为29.4 U/mL。改变发酵条件,有利于聚半乳糖醛酸酶的发酵生产,此时酶活力为基础条件下的5.05倍。     

酱香型大曲中产淀粉酶菌的分离鉴定及发酵特性研究

对酱香型大曲中高产淀粉酶菌株进行分离鉴定,并对其产酶条件和代谢产物进行研究。结合形态学指标和16S rDNA同源序列分析对筛选获得的酱香型大曲中高产淀粉酶菌株进行鉴定,以小麦、麸皮浸提液为液体发酵培养基,研究培养温度、培养基pH值、摇床转速对其产淀粉酶能力的影响。结果显示,该分离菌株为枯草芽孢杆菌（Bacillus Subtilis）,产酶最适条件为：培养温度50 ℃、初始pH值6.0、摇床转速为180 r/min,其酶活力达8 667.79 U/mL。     

微波联合酶法对小麦麸皮品质改良及结构特性影响

本研究以小麦麸皮为原料,采用正交实验优化微波联合酶解对小麦麸皮品质改良工艺参数,并对比分析微波、酶解、微波联合酶解三种处理方式对小麦麸皮结构和性质的影响。研究发现:小麦麸皮微波联合酶解的最佳工艺参数为:微波功率700 W,时间15 min,料水比1:4,木聚糖酶添加量0.4 g,纤维素酶添加量0.4 g,酶解时间4 h,酶解温度60 ℃,此时小麦麸皮中还原糖含量为25.15 mg/mL。小麦麸皮经微波联合酶解处理后,持水性增加了30.18%,植酸含量降低了70.46%,持油性降低了26.69%,脂肪酶(LA)残余酶活降低至6.13%,粗纤维含量降低至2.79%,还原糖含量上升至25.15 mg/mL。傅里叶变换红外光谱分析结果表明微波联合酶解可以破坏分子间的糖苷键,使小麦麸皮细胞壁中纤维素、半纤维素降解,生成小分子的还原糖。扫描电子显微镜观察结果显示微波联合酶解破坏了小麦麸皮结构,使得麸皮表面变粗糙,结构疏松多孔。通过本实验改性的麦麸中脂肪酶残余酶活显著下降,麦麸食用品质明显改善。     

绿色木霉JD-1固态发酵玉米芯产纤维素酶条件优化

以玉米芯与麸皮为主要原料,对影响绿色木霉（Trichoderma viride）JD-1固态发酵的因素如玉米芯与麸皮的比例、氮源浓度、发酵温度、时间、料水比等进行研究。在单因素试验的基础上,采取正交试验设计进行优化。结果表明,最佳固体发酵条件为即玉米芯与麸皮质量比为7∶3,培养温度30 ℃,料液比1∶2.0（g∶mL）,培养时间96 h,接种量为10%。在此优化条件下,羧甲基纤维素酶活力达8.95 IU/g,滤纸酶酶活力达2.00 IU/g。     

花生粕酱油制曲工艺条件优化

以花生粕、小麦麸皮为原料制备花生酱油成曲,研究花生粕与小麦麸皮质量比、米曲霉（Aspergillus oryzae）As3.042接种量、制曲温度和制曲时间4个因素对花生粕酱油成曲中性蛋白酶酶活的影响,并通过单因素试验和响应面试验优化制曲工艺。结果表明,对中性蛋白酶活影响程度从大到小依次为制曲温度＞接种量＞原料质量比＞制曲时间。花生酱油成曲的最佳制曲工艺条件为：花生粕与小麦麸皮质量比8∶2,米曲霉接种量0.65%,制曲温度32 ℃、制曲时间24 h。在此优化条件下,成曲中性蛋白酶活为（2 493.67±7.70） U/g,酸性蛋白酶活为（596.84±23.12） U/g、糖化酶活为（29.91±2.63） U/g、淀粉酶活为（882.85±32.63） U/g、纤维素酶活为（11.72±3.69） U/g。     

Enhanced production of α‐amylase from Bacillus subtilis subsp. spizizenii in solid state fermentation by response surface methodology and its evaluation in the hydrolysis of raw potato starch

This is an attempt to lower the cost of starch hydrolysis by the discovery of new generation α‐amylase. A natural isolate of Bacillus subtilis subsp. spizizenii was capable of producing appreciable amounts of raw potato starch digesting α‐amylase in solid state fermentation of wheat bran. The enzyme productivity has been substantially enhanced by supplementing various nutrients and statistically studying their interactions by response surface methodology. A central composite design for amylase production system elucidated a wheat bran‐based medium supplemented with soybean meal, threonine, and B‐complex vitamins predicting a yield of 521 391 U/g dry solids. The enzyme preparation could effectively digest 5–15% suspension of insoluble potato starch in 6 h revealing the dextrose equivalent of 32–44. The supplementation of a glucoamylase preparation, thereafter, brought about complete saccharification. The yield achieved in the statistically optimized amylase system may be one of the best to date and its capability in directly liquefying raw potato starch granules makes this study novel.     

Alpha‐Cyclodextrin Production using Cyclodextrin Glycosyltransferase from Klebsiella pneumoniae AS‐22

Alpha‐cyclodextrin (α‐CD) production, using cyclodextrin glycosyltransferase (CGTase) from Klebsiella pneumoniae AS‐22, was investigated in the presence of various complexing agents using raw wheat starch and dextrin as substrates. The addition of many alcohols resulted in increased conversion of both raw wheat starch and dextrin to α‐CD. With 125 g/L raw wheat starch, 42.5% (w/w) conversion to CDs was obtained in the presence of 2% (v/v) butan‐1‐ol. The ratio of α:β‐CD formed was 97:3, with negligible formation of γ‐CD and malto‐oligosaccharides. The production of α‐CD was optimized using two‐level factorial designs in three variables: dextrin, hexan‐1‐ol and enzyme concentrations. Under optimum conditions, 12.1% (w/w) conversion of 500 g/L dextrin to total‐CDs was achieved. The ratio of α:β:γ‐CD formed was 91:3:6, with negligible production of malto‐oligosaccharides. This CGTase was strongly inhibited by α:‐CD; 50% inhibition of reaction rate was observed at an initial concentration of 4 g/L α:‐CD. The effectiveness of an ultrafiltration membrane bioreactor for continuous removal of product was also tested.     

Effects of enzymes in fibre-enriched baking

The aim of the present study was to improve the quality of fibre-enriched wheat breads by enzymic treatment of the fibre fraction. The suitability of different enzymes in fibre-enriched baking and their effects on the dietary fibre content and the ratio of insoluble: soluble fibre content of the breads were studied. The enzyme preparations used were a hemicellulolytic culture filtrate of Trichoderma reesei, a specific (pI 9) xylanase of T reesei and Fermizyme, an α-amylase preparation containing a standardised level of hemicellulase activity. Rye bran was extracted in water (10% (w/w) suspension) to determine the solubilities of the β-glucans and pentosans. Addition of T reesei culture filtrate significantly increased the amount of extractable pentosan obtained from nonautoclaved rye bran. Rye bran supplementation (5%) of wheat flour increased the farinograph absorption and dough development time, but had little or no effect on stability and softening of the dough. The added enzymes decreased dough stability and increased softening. Addition of enzymes caused significant differences in the stickiness of the wheat doughs both with (P<0·003) and without (P<0·001) rye bran. Fermizyme significantly increased the stickiness of wheat doughs both with and without rye bran. The baking results of the fibre-enriched breads were improved by the added enzymes. Addition of T reesei culture filtrate increased the specific volume of the wheat breads both with and without rye bran by almost 20%. Enzyme mixtures were more efficient than individual xylanase in softening the bread crumb and reducing the staling rate of wheat breads both with and without rye bran. Incorporation of enzymes reduced the total dietary fibre content of the breads, but at least doubled the amount of soluble pentosan. The proportions of fluorescent cell walls in the breads were detected by microscopical image analysis. Enzyme addition caused the surface area of insoluble cell walls originating from wheat flours to decrease, suggesting that the enzymes exert more effects on wheat endorsperm cell walls than on bran particles. © 1998 SCI.