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解析来自火山环境的嗜热菌的脂肪酶活性。

Unraveling the lipolytic activity of thermophilic bacteria isolated from a volcanic environment.

机构信息

Microbiology Group, Sector of Botany, Department of Biology, National and Kapodistrian University of Athens, Zografou Campus, Zografou 15784, Attica, Greece.

出版信息

Biomed Res Int. 2013;2013:703130. doi: 10.1155/2013/703130. Epub 2013 May 8.

Abstract

In a bioprospecting effort towards novel thermostable lipases, we assessed the lipolytic profile of 101 bacterial strains isolated from the volcanic area of Santorini, Aegean Sea, Greece. Screening of lipase activity was performed both in agar plates and liquid cultures using olive oil as carbon source. Significant differences were observed between the two screening methods with no clear correlation between them. While the percentage of lipase producing strains identified in agar plates was only 17%, lipolytic activity in liquid culture supernatants was detected for 74% of them. Nine strains exhibiting elevated extracellular lipase activities were selected for lipase production and biochemical characterization. The majority of lipase producers revealed high phylogenetic similarity with Geobacillus species and related genera, whilst one of them was identified as Aneurinibacillus sp. Lipase biosynthesis strongly depended on the carbon source that supplemented the culture medium. Olive oil induced lipase production in all strains, but maximum enzyme yields for some of the strains were also obtained with Tween-80, mineral oil, and glycerol. Partially purified lipases revealed optimal activity at 70-80°C and pH 8-9. Extensive thermal stability studies revealed marked thermostability for the majority of the lipases as well as a two-step thermal deactivation pattern.

摘要

在一项针对新型耐热脂肪酶的生物勘探努力中,我们评估了从希腊爱琴海圣托里尼火山地区分离的 101 株细菌的脂肪酶谱。使用橄榄油作为碳源,在琼脂平板和液体培养物中进行脂肪酶活性筛选。两种筛选方法之间观察到显著差异,它们之间没有明显的相关性。虽然在琼脂平板上鉴定出的产脂肪酶菌株的比例仅为 17%,但在液体培养上清液中检测到 74%的菌株具有脂肪酶活性。选择了 9 株表现出较高胞外脂肪酶活性的菌株进行脂肪酶生产和生化特性分析。大多数产脂肪酶菌与地杆菌属和相关属具有高度的系统发育相似性,而其中一株被鉴定为 Aneurinibacillus sp. 脂肪酶的生物合成强烈依赖于补充培养基的碳源。所有菌株都能诱导脂肪酶的产生,但有些菌株在添加吐温-80、矿物油和甘油的培养基中也能获得最大的酶产量。部分纯化的脂肪酶在 70-80°C 和 pH8-9 时表现出最佳活性。广泛的热稳定性研究表明,大多数脂肪酶具有显著的热稳定性,以及两步热失活动力学模式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c221/3662197/fefbe4782fb5/BMRI2013-703130.001.jpg

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