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铜绿假单胞菌 JCM5962(T) 的细胞外脂肪酶:分离、鉴定和特性研究。

Extracellular lipase from Pseudomonas aeruginosa JCM5962(T): Isolation, identification, and characterization.

机构信息

Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Malhaur, Gomti Nagar Extension, Lucknow, India.

出版信息

Int Microbiol. 2018 Dec;21(4):197-205. doi: 10.1007/s10123-018-0016-z. Epub 2018 Jul 10.

Abstract

The study was done to isolate, identify, and characterize a good lipolytic strain from soil. Lipolytic strain isolation was done using tributyrin agar medium. The biochemical testing and 16S rRNA gene sequencing analysis was done for identification. The enzyme was purified using ammonium sulfate precipitation and column chromatography. Results have shown a novel high lipolytic strain of P. aeruginosa JCM5962(T), isolated from soil of sugarcane field. The 16S rRNA sequence analysis confirmed the strain as P. aeruginosa JCM5962(T); further, the sequence was submitted to Genbank (KX946966.1). The isolate produced an extracellular lipase which was purified as single band of 31 kDa. Maximum lipase activity was observed at 50 °C and pH 8.0. Activity was enhanced in the presence of cobalt and benzene solvent, whereas mercury, sodium dodecyl sulfate, and chloroform inhibited it. The enzyme's marked stability and activity at high temperature, alkaline pH and organic solvents suggest that this can be effectively used in a variety of applications in industries and as biotechnological tools.

摘要

本研究旨在从土壤中分离、鉴定和表征一种良好的脂肪酶产生菌。采用吐温 80 琼脂培养基进行脂肪酶产生菌的分离。通过生化测试和 16S rRNA 基因测序分析进行鉴定。采用硫酸铵沉淀和柱层析法对酶进行纯化。结果表明,从甘蔗地土壤中分离到一株新型的高脂肪酶产生菌铜绿假单胞菌 JCM5962(T)。16S rRNA 序列分析证实该菌株为铜绿假单胞菌 JCM5962(T);进一步将序列提交给 Genbank(KX946966.1)。该分离株产生一种胞外脂肪酶,经纯化后为单一条带,分子量为 31 kDa。在 50°C 和 pH 8.0 时观察到最大脂肪酶活性。钴和苯溶剂存在时活性增强,而汞、十二烷基硫酸钠和三氯甲烷则抑制其活性。该酶在高温、碱性 pH 和有机溶剂中的显著稳定性和活性表明,它可以有效地应用于工业中的各种应用和生物技术工具。

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