从睡茄植物中提取的高稳定性 Cu/Zn 超氧化物歧化酶：基因克隆、表达及重组蛋白的特性分析。

A highly stable Cu/Zn superoxide dismutase from Withania somnifera plant: gene cloning, expression and characterization of the recombinant protein.

机构信息

National Botanical Research Institute, Council of Scientific and Industrial Research, Rana Pratap Marg, Lucknow, UP 226001, India.

出版信息

Biotechnol Lett. 2011 Oct;33(10):2057-63. doi: 10.1007/s10529-011-0670-0. Epub 2011 Jun 22.

DOI:10.1007/s10529-011-0670-0

PMID:21695487

Abstract

A gene from Withania somnifera (winter cherry), encoding a highly stable chloroplastic Cu/Zn superoxide dismutase (SOD), was cloned and expressed in Escherichia coli. The recombinant enzyme (specific activity of ~4,200 U mg(-1)) was purified and characterized. It retained ~90 and ~70% residual activities after 1 h at 80 and 95 °C, respectively. At 95 °C, thermal inactivation rate constant (K (d)) of the enzyme was 2.46 × 10(-3) min(-1) and half-life of heat inactivation was 4.68 h. The enzyme was stable against a broad pH range (2.5-11.0). It also showed a high degree of resistance to detergent, ethanol and protease digestion. This recombinant Cu/Zn SOD could therefore have useful applications.

摘要

从冬樱桃（Withania somnifera）中克隆并在大肠杆菌中表达了一种编码高度稳定的叶绿体 Cu/Zn 超氧化物歧化酶（SOD）的基因。该重组酶（比活性约为 4200 U mg(-1)）被纯化并进行了表征。它在 80 和 95°C 下分别保持约 90%和 70%的残余活性。在 95°C 下，该酶的热失活动力学常数（K (d)）为 2.46×10(-3) min(-1)，热失活半衰期为 4.68 h。该酶在较宽的 pH 范围（2.5-11.0）下稳定。它还对去污剂、乙醇和蛋白酶消化具有很高的抗性。因此，这种重组 Cu/Zn SOD 可能具有有用的应用。