Isolation of an alkane-degrading Alcanivorax sp strain 2B5 and cloning of the alkB gene
Isolation of an alkane-degrading Alcanivorax sp strain 2B5 and cloning of the alkB gene
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DOI：
10.1016/j.biortech.2009.08.028
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发表时间：
2010-01-01
影响因子：
11.4
通讯作者：
Shen, Biao
中科院分区：
文献类型：
--
作者：
Liu, Yi-Chen;Li, Ling-Zhi;Shen, Biao
从中国受原油污染的东海地区海泥中分离得到一株革兰氏阴性球菌，菌株2135，经鉴定为Alcanivorax sp.。根据其生理特征并分析其16S rRNA基因序列。菌株2B5能够从作为唯一碳源的原油中降解C-13-C-30正构烷烃和支链烷烃（降植烷和植烷）。菌株2135生长和十八烷降解的最适温度和pH分别为30-37℃和pH 6.0-7.0。菌株 2B5 生长需要 NaCl，并且十八烷降解在 NaCl 浓度为 20 至 90 g L-1 时最高。通过自形成接头PCR（SEFA-PCR）获得的新型烷烃羟化酶基因（alkB）与Alcanivorax borkumensis SK2的alkB1显示出41.9%的推导氨基酸序列同一性。 alkB 基因的功能性异源表达在荧光假单胞菌 KOB2 Delta 1 中实现。含有 Alcanivorax sp. alkB 基因的重组假单胞菌。菌株2135恢复了在C-14和C-16正烷烃上生长的能力。 RT-PCR分析表明alkB基因的表达受到十八烷的诱导。 Alcanivorax sp. 中可能存在其他烷烃羟化酶。第2135章 可能参与短链烷烃和长链烷烃的降解。 (C) 2009 Elsevier Ltd. 保留所有权利。
A Gram-negative coccus, strain 2135, was isolated from the sea mud of the crude oil-polluted Donghai area in China and identified as Alcanivorax sp. based on its physiological characteristics and analysis of its 16S rRNA gene sequence. Strain 2B5 was able to degrade C-13-C-30 n-alkanes and branched alkanes (pristane and phytane) from crude oil as the sole carbon source. The optimal temperature and pH for strain 2135 growth and octadecane degradation were 30-37 degrees C and pH 6.0-7.0, respectively. NaCl is required for strain 2B5 growth and octadecane degradation were highest at NaCl concentrations of 20 to 90 g L-1. A novel alkane hydroxylase gene (alkB), obtained by self-formed adaptor PCR (SEFA-PCR), displays 41.9% deduced amino acid sequence identity with alkB1 of Alcanivorax borkumensis SK2. Functional heterologous expression of the alkB gene was achieved in Pseudomonas fluorescens KOB2 Delta 1. Recombinant Pseudomonas containing the alkB gene of Alcanivorax sp. strain 2135 recovered the ability to grow on C-14 and C-16 n-alkanes. RT-PCR analysis showed that expression of the alkB gene was induced by octadecane. There could be other alkane hydroxylases in Alcanivorax sp. 2135 that could be involved in the degradation of short- and long-chain alkanes. (C) 2009 Elsevier Ltd. All rights reserved.