Post Graduate Department of Biochemistry and Biotechnology, Cell and Molecular Therapeutics Laboratory, Oriental Institute of Science and Technology, Vidyasagar University, Midnapore-721102, West Bengal, India.
Curr Pharm Biotechnol. 2020;21(9):872-881. doi: 10.2174/1389201021666200130113022.
Amylases enzymes hydrolyze starch molecules to produce diverse products including dextrins, and progressively smaller polymers. These include glucose units linked through α-1- 1, α-1-4, α-1-6, glycosidic bonds.
This enzyme carrying an (α /β) 8 or TIM barrel structure is also produced containing the catalytic site residues. These groups of enzymes possess four conserved regions in their primary sequence. In the Carbohydrate-Degrading Enzyme (CAZy) database, α-amylases are classified into different Glycoside Hydrolase Families (GHF) based on their amino acid sequence. The present objective was to study one such enzyme based on its molecular characterization after purification in our laboratory. Its main property of solid-natural starch degradation was extensively investigated for its pharmaceutical/ industrial applications.
Amylase producing bacteria Bacillus cereus sm-sr14 (Accession no. KM251578.1) was purified to homogeneity on a Seralose 6B-150 gel-matrix and gave a single peak during HPLC. MALDITOF mass-spectrometry with bioinformatics studies revealed its significant similarity to α/β hydrolase family. The enzyme showed an efficient application; favourable Km, Vmax and Kcat during the catalysis of different natural solid starch materials. Analysis for hydrolytic product showed that this enzyme can be classified as the exo-amylase asit produced a significant amount of glucose.
Besides the purified enzyme, the present organism Bacillus cereus sm-sr14 could degrade natural solid starch materials like potato and rice up to the application level in the pharmaceutical/ industrial field for alcohol production.
淀粉酶酶将淀粉分子水解产生多种产物，包括糊精和逐渐更小的聚合物。这些包括通过α-1-1、α-1-4、α-1-6、糖苷键连接的葡萄糖单元。
这种具有（α/β）8 或 TIM 桶结构的酶也含有催化部位残基。这些酶组在其一级序列中具有四个保守区域。在碳水化合物降解酶（CAZy）数据库中，α-淀粉酶根据其氨基酸序列被分类为不同的糖苷水解酶家族（GHF）。本研究的目的是在实验室中对一种酶进行纯化后，根据其分子特征进行研究。其对天然淀粉的降解特性已广泛研究，以应用于制药/工业。
产酶细菌蜡状芽孢杆菌 sm-sr14（登录号 KM251578.1）在 Seralose 6B-150 凝胶基质上纯化为均相，在 HPLC 中呈现单一峰。MALDITOF 质谱与生物信息学研究表明，它与α/β水解酶家族具有显著的相似性。该酶在不同天然固体淀粉材料的催化中表现出高效的应用，具有有利的 Km、Vmax 和 Kcat。水解产物分析表明，该酶可归类为外切淀粉酶，因为它产生了大量的葡萄糖。
除了纯化酶之外，本研究中的蜡状芽孢杆菌 sm-sr14 还可以降解天然固体淀粉材料，如土豆和大米，达到制药/工业领域生产酒精的应用水平。
