Mode of sugar phosphorylation inClostridium thermocellum

Fungi were screened for important industrial enzymes produced from industrial chips and sawdust of laurel (Louro inamui, Ocotea cymbarum) and cedar (Cedro, Cedrella odorata). Seven hyphomycetes and one zygomycete were isolated and characterized. Two different media for testing the enzymatic activities were used. In general, in potato dextrose (1%) medium (M-3) a preponderancy of ligninolytic over cellulolytic enzymes was observed. In potato infusion-sawdust wood (1%) medium (M-4) the cellulolytic, xylanolytic, and lipolytic enzymes were efficiently induced. Significant modulation of enzyme production by the carbon source was found in the extracted fungi from self-heated industrial chips piles of cedar and laurel trees at the Amazonian region.     

从噬菌体展示七肽肽库筛选α-淀粉酶特异性配体

为了建立蛋白质亲和配基的高效筛选方法，以淀粉酶为靶分子，利用交替洗脱法从七肽噬茵体展示库中筛选具有高亲和力的噬茵体配体．结果表明，交替洗脱法筛选出的特异性配体的回收率和ELISA信号值均优于酸洗脱法；采用交替洗脱法能够更加有效和迅速地筛选到淀粉酶的高亲和力配体．分析筛选得到的不同噬茵体克隆DNA插入片断的氨基酸序列，发现了可能与配体高亲和性有关的氨基酸残基．     

An analytical method for measuring α‐amylase activity in starch‐containing foods

The quality of starch‐containing foods may be significantly impaired by contamination with very small amounts of α‐amylase, which can enzymatically hydrolyze the starch and cause viscosity loss. Thus, for quality control, it is necessary to have an analytical method that can measure low amylase activity. We developed a sensitive analytical method for measuring the activity of α‐amylase (from Bacillus subtilis) in starch‐containing foods. The method consists of six steps: (1) crude extraction of α‐amylase by centrifugation and filtration; (2) α‐amylase purification by desalting and anion‐exchange chromatography; (3) reaction of the purified amylase with boron‐dipyrromethene (BODIPY)‐labeled substrate, which releases a fluorescent fragment upon digestion of the substrate, thus avoiding interference from starch derivatives in the sample; (4) stopping the reaction with acetonitrile; (5) reversed‐phase solid‐phase extraction of the fluorescent substrate to remove contaminating dye and impurities; and (6) separation and measurement of BODIPY fluorescence by HPLC. The proposed method could quantify α‐amylase activities as low as 10 mU/mL, which is enough to reduce the viscosity of starch‐containing foods. Copyright © 2012 John Wiley & Sons, Ltd.     

Evaluation of in vitro,in silico antidiabetic and antioxidant potential of bioactivity based isolated “Pakistanine” from Berberis baluchistanica

Bioassay based fractionation of methanolic extract of Berberis baluchistanica (Berberidaceae), used traditionally for internal injuries, led to the isolation of known compounds (1–4). The structure of these compounds was elucidated by different spectroscopic analysis and available literature data. Antidiabetic and antioxidant potentials of B. baluchistanica fractions and isolated compounds were evaluated using in vitro alpha- amylase and DPPH assays. The isolated compounds were identified as obamegine (1), pakistanine (2), 8-oxyberberine (3) and baluchistine (4). Obamegine was reported from many other species of this genus but it is first time isolated from B. baluchistanica in present study. Moreover, in vitro pakistanine (2) was found as bioactive lead molecule for hypoglycemic (IC₅₀:40.26 µg/ml) and antioxidant (IC₅₀:14.15 µg/ml) activities compared to acarbose (IC₅₀:33.68 µg/ml) and ascorbic acid (IC₅₀:0.41 µg/ml). To the best of our knowledge, no previous data were available for these biological activities. Additionally, in silico antidiabetic and antioxidant activity of pakistanine against two proteins, α-amylase (-9.7 kcal/mol) and tyrosinase (-8.7 kcal/mol) are reported here for the first time. The molecular docking binding interactions authenticate and support the above-mentioned activities and are helpful in predicting the mechanism of action of pakistanine (2).     

米根霉发酵产L-乳酸体系中淀粉酶的研究

米根霉As3.819是一株L-乳酸的高产菌株,且能够分泌淀粉酶直接利用淀粉发酵生产乳酸。文章研究了不同培养基主要成分下米根霉菌株As3.819生产L-乳酸和淀粉酶情况,并对发酵液中淀粉酶水解产物进行分析。研究结果表明,在甘薯淀粉8%、硫酸铵为0.4%时L-乳酸和淀粉酶的产量相对较高,且在培养基中加入麸皮后能增加淀粉酶的分泌,种子培养基中碳源为麦芽糖时也能起到很好的效果。     

黄酒生产菌种分离培养基的筛选

通过大量选择试验 ,确定适合于黄曲霉菌种分离的几种培养基 ,应用于紫外线诱变和自然分离 ,获得较好结果。     

强苯赛对水稻幼苗根系生长的影响

强苯赛浸种对水稻幼苗的根长、根重和根系体积都有促进作用,能提高根尖细胞分裂指数,能使胚根尖切段伸长和发根力增强,并能提高胚乳中淀粉酶活性和根系的呼吸强度。     

嗜盐菌碱性淀粉酶特性研究

碱性嗜盐菌ＨａｌｏｂａｃｔｅｒｉｕｍｓｐＨ－３７１产生的淀粉酶经超滤、硫酸铵沉淀和ＤＥＡＥ－纤维素纯化后，ＳＤＳ－ＰＡＧＥ凝胶电泳为一条带，分子量为２７０００，等电点３．７，最适反应ｐＨ９．０，最适反应温度６５℃．ＮａＣｌ是维持酶活性的必需因子，酶解产物为麦芽糖、麦芽三糖、麦芽四糖和麦芽五糖     

糖化酶发酵液超滤条件的研究

本文以浓差极化—凝胶层理论为基础,研究了影响糖化酶超滤的各种工艺因素,并测定滤饼比阻力,得到滤饼比阻力与压差的函数关系.结果表明在浓差极化现象存在下,透过液通量随操作时间、透过液总体积以及主体溶液浓度的增加而减少,随搅拌速度的增加而增加;当操作压差达一定值后,透过液通量与压差无关.粗酶液经超滤后,可得到高活性浓缩酶.     

试管比色法检测淀粉酶值实验教学中的问题探讨

淀粉酶值是蜂蜜卫生检验中的重要检测项目之一,其活性的高低是衡量蜂蜜成熟度、加热程度和贮存时间的重要指标,而试管比色法被广泛应用于高等院校实验教学中用以检测淀粉酶的活性。针对该方法在实验教学过程中出现的不同问题分别进行了探讨。