Raul Dibyangana, Biswas Tania, Mukhopadhyay Suchita, Kumar Das Shrayan, Gupta Suvroma
Department of Biotechnology, Haldia Institute of Technology, ICARE Complex, Purba Medinipur 721657, India.
Biochem Res Int. 2014;2014:568141. doi: 10.1155/2014/568141. Epub 2014 Jan 12.
Amylase is an enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like use as digestives, for the production of ethanol and high fructose corn syrup, detergents, desiring of textiles, modified starches, hydrolysis of oil-field drilling fluids, and paper recycling. In the present work, solid state fermentation (SSF) for α -amylase production has been used in lieu of submerged fermentation (SmF) due to its simple technique, low capital investment, lower levels of catabolite repression, and better product recovery. Bacillus subtilis has been well known as producer of alpha amylase and was tested using solid state fermentation for 48 hours at 37°C with wheat bran as substrate. Comparison between different fermentation hours demonstrated high yield of alpha amylase after 48 hours. This alpha amylase has optimum pH and temperature at 7.1 and 40°C, respectively. With the goal to purify alpha amylase, 30-70% (NH4)2SO4 cut concentrated the amylase activity threefold with respect to crude fermented extract. This was verified in quantitative DNS assay method as well as in zymogram gel profile. The exact molecular weight of the amylase is yet to be determined with the aid of other protein purification techniques.
淀粉酶是一种催化淀粉分解为糖的酶,在多种领域发挥着关键作用,如用作消化剂、用于生产乙醇和高果糖玉米糖浆、洗涤剂、纺织品加工、变性淀粉、油田钻井液水解以及纸张回收利用等。在本研究中,由于固态发酵(SSF)技术简单、资本投入低、分解代谢物阻遏水平低且产品回收率高,因此被用于替代深层发酵(SmF)来生产α -淀粉酶。枯草芽孢杆菌一直是众所周知的α -淀粉酶生产者,本研究以麸皮为底物,在37°C下使用固态发酵对其进行了48小时的测试。不同发酵时间的比较表明,48小时后α -淀粉酶产量很高。这种α -淀粉酶的最适pH值和温度分别为7.1和40°C。为了纯化α -淀粉酶,用30 - 70%的硫酸铵沉淀浓缩,相对于粗发酵提取物,淀粉酶活性提高了三倍。这在定量DNS测定法以及酶谱凝胶图谱中都得到了验证。淀粉酶的确切分子量还有待借助其他蛋白质纯化技术来确定。