从阿拉伯海沉积物来源的海洋宏基因组文库中克隆和鉴定一种耐盐α-淀粉酶
Molecular cloning and characterization of a halotolerant α-amylase from marine metagenomic library derived from Arabian Sea sediments.
作者信息
Nair Harisree P, Vincent Helvin, Puthusseri Rinu Madhu, Bhat Sarita G
机构信息
Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, 682022, India.
出版信息
3 Biotech. 2017 May;7(1):65. doi: 10.1007/s13205-017-0674-0. Epub 2017 Apr 27.
Abstract
Functional screening of a metagenomic library of marine sediment revealed an amylolytic clone BTM109. This report states the purification and characterization of a moderately halotolerant α-amylase, with more than 51% activity in 2.5 M NaCl. The molecular mass of purified protein was determined to be 55.7 kDa by MALDI-TOF MS. The optimum pH for enzyme activity was pH 7 and temperature for maximal activity was 40 °C. At 5 mM concentration, Ca enhanced the enzyme activity indicating that the enzyme is a Ca dependent α-amylase which was confirmed by the starch hydrolysis pattern using TLC. These physico-chemical properties support the suitability of this enzyme for various industrial applications.
摘要
对海洋沉积物宏基因组文库进行功能筛选，发现了一个淀粉酶解克隆BTM109。本报告阐述了一种中度耐盐α-淀粉酶的纯化及特性，该酶在2.5M NaCl中活性超过51%。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）测定，纯化蛋白的分子量为55.7 kDa。酶活性的最佳pH值为7，最大活性温度为40°C。在5 mM浓度下，Ca增强了酶活性，表明该酶是一种依赖Ca的α-淀粉酶，这通过使用薄层色谱（TLC）的淀粉水解模式得到证实。这些物理化学性质表明该酶适用于各种工业应用。
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