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.     

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).     

聚乙烯醇、聚乙烯胺与淀粉酶相互作用荧光光谱

采用荧光光谱和紫外吸收光谱法研究聚乙烯醇(PEG)和四乙烯五胺(TEPA)与淀粉酶相互作用。结果表明,PEG会增强淀粉酶内源性荧光和酪氨酸残基所处微环境的疏水性;TEPA对淀粉酶内源性荧光的猝灭机制属于动态猝灭,但同时也存在静态猝灭特征,并使色氨酸残基所处微环境的极性增大;在所考察的范围内,PEG与淀粉酶的结合常数在40℃达到最高,TEPA对淀粉酶荧光的动态猝灭结合常数在30℃以上趋于最大,PEG、TEPA与淀粉酶之间的作用力属于疏水与静电作用相结合。     

Pb2+对α-淀粉酶活性的影响及其光谱学研究

在α 淀粉酶介质中加入Pb2 ,通过光谱学手段研究Pb2 对α 淀粉酶活性影响的作用机理。结果表明低浓度的Pb2 对酶有激活作用 ,高浓度则严重抑制酶活性。在高浓度下 ,Pb2 能完全竞争出α 淀粉酶中的Ca2 而结合到了α 淀粉酶上 ,其EXAFS的测试表明Pb2 与氨基酸残基上的羧基氧发生了配位 ,配位数为 2 ,Pb—O键长为 0 2 34nm。圆二色 (CD)谱测试表明 ,高浓度的Pb2 结合使α 淀粉酶的二级结构被破坏 ,α 螺旋含量、β 转角及无规则卷曲大量下降 ,β 折叠、二硫键含量大量增多 ,Pb2 的这种完全结合致使酶的构象改变 ,形成无效的酶 Pb2 底物复合物 ,因而使酶失去活性。     

Behavior of biodegradable composites based on starch reinforced with modified cellulosic fibers

The aims of this study were to develop composite films based on potato starch and cellulose modified with toluenediisocyanate, to investigate their morphology and structure, and to evaluate their behavior to enzymatic hydrolysis and their potential use to manufacture of biodegradable seedling pots. The effects of modified cellulosic fibers upon mechanical properties and biodegradability of composite materials based on starch matrix were investigated by tensile strength tests, Fourier infrared spectroscopy, X‐ray diffraction, and dynamic vapor sorption. The behavior of the films to enzymatic hydrolysis with amylase and cellulase was studied; the kinetic of enzymatic hydrolysis and characterization of materials are reported. Chemical modification of cellulose improves tensile strength with about 47%, and decreases the biodegradability of composites making them more resistant to microbial attack, thus prolonging their shelf life. Copyright © 2015 John Wiley & Sons, Ltd.     

Synthesis,characterization and α‐amylase and α‐glucosidase inhibition studies of novel vanadyl chalcone complexes

A series of chalcone ligands and their corresponding vanadyl complexes of composition [VO (L^I–IV)₂(H₂O)₂]SO₄ (where L^I = 1,3‐Diphenylprop‐2‐en‐1‐one, L^II = 3‐(2‐Hydroxy‐phenyl)‐1‐phenyl‐propenone, L^III = 3‐(3‐Nitro‐phenyl)‐1‐phenyl‐propenone, L^IV = 3‐(4‐Methoxy‐phenyl)‐1‐phenyl‐propenone) have been synthesized and characterized using various spectroscopic (Fourier‐transform infrared, electrospray ionization mass, nuclear magnetic resonance, electron paramagnetic resonance, thermogravimetric analysis, vibrating sample magnetometer) and physico‐analytic techniques. Antidiabetic activities of synthesized complexes along with chalcones were evaluated by performing in vitro and in silico α‐amylase and α‐glucosidase inhibition studies. The obtained results displayed moderate to significant inhibition activity against both the enzymes by vanadyl chalcone complexes. The most potent complexes were further investigated for the enzyme kinetic studies and displayed the mixed inhibition for both the enzymes. Further, antioxidant activity of vanadyl chalcone complexes was evaluated for their efficiency to release oxidative stress using 2,2‐diphenyl‐1‐picryl‐hydrazyl‐hydrate assay, and two complexes (Complexes 2 and 4 ) have demonstrated remarkable antioxidant activity. All the complexes were found to possess promising antidiabetic and antioxidant potential.     

Studies on the Thermodenaturation Behavior of Bacillus subtilis α‐Amylase on Chromatographic Media

The thermodenaturation behavior of Bacillus subtilis α‐amylase on some chromatographic media was studied by determining their adsorption parameters with frontal analysis. The experimental results show that on a RP‐C₁₈ reversed‐phase medium, a Chelating Sepharose Fast‐Flow chelated by Zn²⁺ affinity medium and a WCX‐1 cation‐exchange medium, a stable conformation of α‐amylase molecule separately exists below or over 30 °C; while on a PEG‐400 hydrophobic medium and a modified PEG‐400 medium, a stable conformation of α‐amylase molecule separately exists below 40 and 30 °C, and when the experimental temperatures are separately over 40 and 30 °C, a drastically conformational change of α‐amylase molecules can continuously take place. And by combining the intrinsic fluorescence emission spectrum and thermal inactivation profile of α‐amylase in free solution and on the PEG‐400 and modified PEG‐400 hydrophobic media, it can be concluded that in liquid chromatographic procedure, chromatographic media can induce the conformational change of α‐amylase molecules and promote their thermodenaturation; and in hydrophobic interaction chromatography, the higher the hydrophobicity of chromatographic medium, the lower the conformational change temperature of α‐amylase molecules on the chromatographic medium.     

黑鱾（Girellaleonina）淀粉酶与蛋白酶的活力研究

通过测定1＋龄黑鱾的胃、盲囊、肝脏和肠道4个消化组织中蛋白酶和淀粉酶活性,并研究其与温度及pH值的关系,可以为黑纪的合理投饵提供理论依据．实验设9个温度梯度（10～50℃）和14个pH缓冲液梯度（pH2．0～9．0）,用淀粉-碘显色法测淀粉酶活力,用福林-酚试剂法测蛋白酶活力．结果表明：黑纪盲囊、肠道中淀粉酶和蛋白酶活力均较高,最适温度为35℃,淀粉酶最高活力为每克鲜组织46．02U和42．61U蛋白酶最高活力为每克鲜组织156．52u和147．79U;胃中酶的最适温度为40℃,肝脏中淀粉酶温度要求稍低,最适温度为30℃,酶活力值也最低;胃淀粉酶和胃蛋白酶最适pH值分别为6．5和3．5,盲囊、肠道和肝脏组织中的最适pH值相近,为7．0-8．0．可以认为黑鲍胃中主要存在着酸性蛋白酶,胃是食物蛋白质初步分解消化的场所;Q10值充分反映了温度对消化酶活力的影响,当温度从10℃上升至20℃时。2种消化酶活力增幅显著,可作为开始投饵的温度指标．     

直链淀粉手性固定相高效液相色谱法拆分比索洛尔对映体

采用直链淀粉手性固定相高效液相色谱法在正相条件下直接拆分了比索洛尔对映异构体。分别以异丙醇、乙醇为有机改性剂,考察了流动相的组成与配比、流速及柱温等因素对比索洛尔对映体分离的影响。确定了比索洛尔对映体的最佳拆分条件:流动相正己烷-乙醇-二乙胺(体积比为88∶12∶0.1),流速0.6 mL/min,检测波长270 nm,柱温20 ℃。该方法可快捷、简便地拆分比索洛尔对映体。