酶法制备花生粕醒酒多肽

为更合理有效地利用花生粕资源,制备具有醒酒作用的花生粕多肽,本实验以Alcalase AF 2.4L蛋白酶酶法制备花生粕醒酒肽,经分级分离后,通过体外和动物实验验证其醒酒效果。结果表明,花生粕醒酒肽制备最佳工艺条件为：酶解时间3 h、酶解温度35 ℃、pH 9.5及料液比1∶30（m/V）。该条件下乙醇脱氢酶（alcoholdehydrogenase,ADH）激活率及多肽得率均较高。经分级分离后,分子质量在1 000～3 000 D的多肽对ADH激活作用最强,激活率为30.47%。G25凝胶色谱分析表明,花生粕多肽中小于3 000 D的多肽占89.55%,ADH激活率为29.25%。动物实验证明,花生粕多肽对小鼠有显著的防醉醒酒作用。小鼠血液乙醇含量测定结果表明,高剂量的花生粕多肽在60～90 min内能显著降低小鼠血液中的乙醇含量。     

B.subtilis蛋白酶高产菌株的筛选与鉴定

本文以中性蛋白酶生产菌Ｂ．ｓｕｂｔｉｌｉｓＡＳ１．３９８为出发菌，用ＮＴＧ与紫外辐照相结合的方法对其进行复合诱变，筛选到一株蛋白酶活性明显高于出发菌的正突变株，命名为Ｂ．ｓｕｂｔｉｌｉｓＭＬ。通过对该突变株蛋白酶活性曲线的绘制、酶作用最适ＰＨ的确定、所产一的酶性质的分析及比酶活性的测定，发现该突变株扩酶活性比ＡＳ１．３９８至少高出３０％，最适反应ＰＨ７－８，酶活性受ＥＤＴＡ的强烈抑制，其突变位点很     

金线鱼肝胰脏蛋白酶激活提取条件的优化

用响应面法对金线鱼肝胰脏蛋白酶的激活及提取条件进行了优化。比较了不同激活剂的激活效果。研究了提取时间和pH值交互作用对蛋白酶活力的影响。结果表明:肠提液与CaCl2协同作为激活剂的激活效果最好。优化后激活时间为6 h,激活温度为27℃,肠提液质量分数为7%,CaCl2浓度为5.20 mmol/L,酶活力为1578.14 U/(g.min)。肝胰脏蛋白酶最佳提取条件为:提取时间8.5 h,提取温度11℃,pH 5.4,酶活力为1431 U/(g.min)。蛋白酶复性电泳结果说明酶活力增加显著。     

不同酶对藜麦蛋白肽制备的影响及其抗氧化活性研究

本实验以藜麦蛋白为原料,采用碱性蛋白酶、复合蛋白酶、风味蛋白酶、木瓜蛋白酶、中性蛋白酶分别对藜麦蛋白进行水解,制备藜麦蛋白肽。通过单因素实验对五种酶水解制备的藜麦蛋白肽以蛋白水解度、DPPH自由基清除能力、·OH自由基清除能力、ABTS+自由基清除能力、·O2-自由基清除能力为测定指标,研究五种蛋白酶对藜麦蛋白的水解能力和抗氧化活性影响,选出效果相对较佳的酶水解工艺。综合各指标的结果表明,碱性蛋白酶和复合蛋白酶水解能力较强,制备的藜麦蛋白肽抗氧化活性较其他两种酶更好。碱性蛋白酶和复合蛋白酶最佳酶解工艺分别为:酶与底物比0.5%(w/w),底物与水为1∶25(w/v),水解温度50℃,水解时间5h,pH10.0;酶与底物比0.5%(w/w),底物与水为1∶25(w/v),水解温度55℃,水解时间5h,pH8.0。本研究为藜麦蛋白的后期的深加工利用提供一定理论依据。     

蛋白酶的生产和应用(上)

为充分了解蛋白酶,该文从蛋白酶的分类与底物专一性、微生物蛋白酶的生产等方面介绍了蛋白酶。蛋白酶按活性中心可分为丝氨酸蛋白酶、天门冬氨酸蛋白酶、半胱氨酸蛋白酶和金属蛋白酶;按最适pH值来分又分酸性蛋白酶、中性蛋白酶和碱性蛋白酶。微生物蛋白酶的产生与微生物生长有关,合成受分解代谢阻遏的调控和底物厦其类似物的诱导。并介绍了生产蛋白酶的微生物。     

Improving the performance of industrial ethanol‐producing yeast by expressing the aspartyl protease on the cell surface

The yeasts used in fuel ethanol manufacture are unable to metabolize soluble proteins. The PEP4 gene, encoding a vacuolar aspartyl protease in Saccharomyces cerevisiae, was either secretively or cell‐surface anchored expressed in industrial ethanol‐producing S. cerevisiae. The obtained recombinant strains APA (expressing the protease secretively) and APB (expressing the protease on the cell wall) were studied under ethanol fermentation conditions in feed barley cultures. The effects of expression of the protease on product formation, growth and cell protein content were measured. The biomass yield of the wild‐type was clearly lower than that of the recombinant strains (0.578 ± 0.12 g biomass/g glucose for APA and 0.582 ± 0.08 g biomass/g glucose for APB). In addition, nearly 98–99% of the theoretical maximum level of ethanol yield was achieved (relative to the amount of substrate consumed) for the recombinant strains, while limiting the nitrogen source resulted in dissatisfactory fermentation for the wild‐type and more than 30 g/l residual sugar was detected at the end of fermentation. In addition, higher growth rate, viability and lower yields of byproducts such as glycerol and pyruvic acid for recombinant strains were observed. Expressing acid protease can be expected to lead to a significant increase in ethanol productivity. Copyright © 2010 John Wiley & Sons, Ltd.     

The effect of limited proteolysis by different proteases on the formation of whey protein fibrils

Four proteases: trypsin, protease A, pepsin, and protease M were selected to modify whey protein concentrate (WPC) at a low degree of hydrolysis (0.1, 0.2, and 0.3%) before adjusting to pH 2.0 and heating at 90°C to gain insight into the influence of proteolysis on fibril formation. The kinetics of fibril formation were performed on native and modified WPC using the fluorescent dye thioflavin T in conjunction with transmission electron microscopy and far-UV circular dichroism spectroscopy for the morphological and secondary structural analyses. The change in surface hydrophobicity and content of free sulfhydryl groups were also observed during the formation of fibrils for the native and modified WPC. The content of aggregation and thioflavin T kinetic data indicated that the ability of fibril formation was apparently different for WPC modified by the 4 proteases. Whey protein concentrate modified by trypsin aggregated more during heating and the fibril formation rate was faster than that of the native WPC. Whey protein concentrate modified by the other proteases showed slower aggregation with worse amyloid fibril morphology. Compared with the native WPC, the structure of WPC changed differently after being modified by proteases. The state of α-helix structure for modified WPC played the most important role in the formation of fibrils. Under the mild conditions used in this work, the α-helix structure of WPC modified by trypsin caused little destruction and resulted in fibrils with good morphology; the content of α-helices for WPC modified by other proteases decreased to 36.19 to 50.94%; thus, fibril formation was inhibited. In addition, it was beneficial for the modified WPC to form fibrils such that the surface hydrophobicity increased and the content of free sulfhydryl groups slightly decreased during heating.     

Enzymatic treatment of wool fabrics - opportunity of the improvement on some physical and chemical properties of the fabrics

The enzymatic treatment of textiles significantly improves some of their physicochemical properties as well as increases their aesthetic values and comfort of use. Enzymes are used in order to develop environmentally friendly processes by reducing the concentration of chemical agents, water and energy consumption. In the present study, an attempt was made to treat the wool fabric with different concentrations (1, 3, and 5 g/L) of protease enzyme and observed the effects on physical and chemical properties including softness, absorbency, pilling resistance, weight loss, tensile strength loss, water retention, felting shrinkage, alkali solubility and urea-bisulphite solubility of wool fabric. The results of pretreated and enzyme-treated samples are compared to those obtained for untreated wool fabric. Enzyme-treated wool fabrics showed improvement in softness, absorbency, pilling resistance and felting shrinkage and a slight increase in weight loss, tensile strength loss, alkali solubility and urea-bisulphite solubility, and decrease in water retention of the fabric.     

Effect of decortication and protease treatment on the kinetics of liquefaction,saccharification, and ethanol production from sorghum

BACKGROUND: This study analyzes the effect of decortication and protease treatment on the kinetics of liquefaction, saccharification and ethanol production from sorghum kernels. In general, bioethanol yields from sorghum are lower than those from maize. This has been attributed to reduced access of starch‐degrading enzymes due to the crosslinked protein net in the sorghum kernels. RESULTS: Liquefaction is described as a zero order kinetics process, with reaction rates enhanced by protease treatment. The use of protease almost doubled the liquefaction rate in both whole and decorticated sorghum, compared with untreated kernels. During saccharification of decorticated sorghum, protease treatment significantly affected the glucose/starch yield and the glucose concentration profile over time. When compared with maize, protease treatment of decorticated sorghum resulted in superior ethanol production rates. Specific ethanol yields during fermentation were statistically comparable with those for maize. CONCLUSION: Protease treatment of decorticated sorghum kernels can impart substantial economic benefits in terms of improvement of bioethanol yield (13% over whole sorghum) and in reduced fermentation time (approximately 50% with respect to maize). Copyright © 2010 Society of Chemical Industry