从奇利卡湖分离的芽孢杆菌 SP-CH7 中提取的去污稳定碱性 α-淀粉酶 AA7 的探索。
Exploration of detergent-stable alkaline α-amylase AA7 from Bacillus sp strain SP-CH7 isolated from Chilika Lake.
机构信息
Department of Microbiology, C.B.S.H., Orissa University of Agriculture and Technology, Unit 7, Surya Nagar, Bhubaneshwar, Odisha 751003, India.
Department of Microbiology, C.B.S.H., Orissa University of Agriculture and Technology, Unit 7, Surya Nagar, Bhubaneshwar, Odisha 751003, India.
出版信息
Int J Biol Macromol. 2019 Nov 1;140:825-832. doi: 10.1016/j.ijbiomac.2019.08.006. Epub 2019 Aug 8.
An alkali-tolerant bacterial isolate CH7 was isolated from Chilika Lake sediments that produced α-amylase at high pH. It showed >15 mm zone diameter at pH 11.0 on agar plate containing 1% starch hence considered for study. It survived up to 10% NaCl concentration on nutrient agar plates. Taxonomically it was identified as Bacillus sp. strain SP-CH7 having GenBank Accession No. KU761266. Optimized alkaline α-amylase, AA7 showed 202.857 specific activity (IU/mg), 65.95 purification fold and 25.94 yield (%) respectively. Molecular weight was revealed by SDS-PAGE as 55 kDa. The K value was 0.688 mg/mL and so was 2.342 IU/mL V obtained for AA7 towards starch. This enzyme showed 92.79% and 91.14% stability at pH 11.0 and 13.0 respectively and up to 75 °C stability was also noted. Enzyme activity was stimulated by Co, Na and Ca. It was stable in presence of EDTA, SDS, all the powdered detergents as well as with liquid detergent "Ezee" which vouches for its potential application in Detergent industries.
从奇利卡湖沉积物中分离到一株耐碱细菌 CH7，该菌在高 pH 值下产生α-淀粉酶。在含有 1%淀粉的琼脂平板上，其产生的酶活圈直径>15mm，因此被认为具有研究价值。该菌在含 10%NaCl 的营养琼脂平板上仍能存活。基于系统发育分析，该菌被鉴定为芽孢杆菌属（Bacillus sp.）菌株 SP-CH7，其基因序列已提交至 GenBank，登录号为 KU761266。优化后的碱性α-淀粉酶 AA7 的比酶活为 202.857IU/mg，纯化倍数为 65.95，酶活回收率为 25.94%。SDS-PAGE 结果表明其分子量约为 55kDa。AA7 对淀粉的 K m 值和 V max 值分别为 0.688mg/mL 和 2.342IU/mL。该酶在 pH 11.0 和 13.0 时的稳定性分别为 92.79%和 91.14%，在 75°C 下也具有稳定性。该酶的活性可被 Co、Na 和 Ca 激活。EDTA、SDS、各种粉状洗涤剂以及液体洗涤剂“Ezee”均对其活性无影响，表明其在洗涤剂工业中有潜在的应用价值。
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