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来自安全芽孢杆菌DVL-43的经统计学增强的新型有机溶剂稳定脂肪酶的应用。

Application of a statistically enhanced, novel, organic solvent stable lipase from Bacillus safensis DVL-43.

作者信息

Kumar Davender, Parshad Rajinder, Gupta Vijay Kumar

机构信息

Department of Biochemistry, Kurukshetra University, Kurukshetra 136119, India.

Indian Institute of Integrative Medicine, Jammu 180001, India.

出版信息

Int J Biol Macromol. 2014 May;66:97-107. doi: 10.1016/j.ijbiomac.2014.02.015. Epub 2014 Feb 15.

Abstract

This paper presents the molecular identification of a newly isolated bacterial strain producing a novel and organic solvent stable lipase, statistical optimization of fermentation medium, and its application in the synthesis of ethyl laurate. On the basis of nucleotide homology and phylogenetic analysis of 16S rDNA sequence, the strain was identified as Bacillus safensis DVL-43 (Gen-bank accession number KC156603). Optimization of fermentation medium using Plackett-Burman design and response surface methodology led to 11.4-fold increase in lipase production. The lipase from B. safensis DVL-43 exhibited excellent stability in various organic solvents. The enzyme retained 100% activity after 24h incubation in xylene, DMSO and toluene, each solvent being used at a concentration of 25% (v/v). The use of partially purified DVL-43 lipase as catalyst in the synthesis of ethyl laurate, an esterification product of lauric acid and ethanol, resulted in 80% esterification in 12h under optimized conditions. The formation of ethyl laurate was confirmed using TLC and (1)H NMR. Organic solvent stable lipases exhibiting potential application in enzymatic esterification are in great demand in flavor, fine chemicals and pharma industries. We could not find any report on lipase production from B. safensis strain and its application in esterification.

摘要

本文介绍了一种新分离的产新型且对有机溶剂稳定的脂肪酶的细菌菌株的分子鉴定、发酵培养基的统计优化及其在月桂酸乙酯合成中的应用。基于16S rDNA序列的核苷酸同源性和系统发育分析,该菌株被鉴定为安全芽孢杆菌DVL-43(Gen-bank登录号KC156603)。使用Plackett-Burman设计和响应面方法对发酵培养基进行优化,使脂肪酶产量提高了11.4倍。安全芽孢杆菌DVL-43产生的脂肪酶在各种有机溶剂中表现出优异的稳定性。在二甲苯、二甲基亚砜和甲苯中,每种溶剂浓度为25%(v/v),孵育24小时后,该酶仍保留100%的活性。使用部分纯化的DVL-43脂肪酶作为催化剂合成月桂酸乙酯(月桂酸和乙醇的酯化产物),在优化条件下12小时内酯化率达80%。使用薄层色谱法和氢核磁共振波谱法确认了月桂酸乙酯的形成。在香料、精细化学品和制药行业中,对在酶促酯化中具有潜在应用价值的有机溶剂稳定脂肪酶有很大需求。我们未找到任何关于安全芽孢杆菌菌株产脂肪酶及其在酯化中的应用的报道。

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