Ji Nannan, Wang Xiuli, Yin Chong, Peng Wanli, Liang Rubing
State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.
Front Microbiol. 2019 Mar 12;10:400. doi: 10.3389/fmicb.2019.00400. eCollection 2019.
AlkB monooxygenases in bacteria are responsible for the hydroxylation of medium- and long-chain -alkanes. In this study, one CrgA protein of SJTD-1, a member of LysR family, was proved to regulate AlkB2 monooxygenase and the degradation of medium-to-long-chain -alkanes (C-C) by directly binding to the upstream of 2 gene. Two specific sites for CrgA binding were found in the promoter region of 2 gene, and the imperfect mirror repeat (IIR) structure was proved critical for CrgA recognition and binding. Hexadecyl CoA and octadecyl CoA could effectively release the CrgA binding and start the transcription of gene, implying a positive regulation of metabolic intermediate. In the presence of medium-to-long-chain -alkanes (C-C), deletion of gene could enhance the transcription and expression of AlkB2 monooxygenase significantly; and in n-octadecane culture, strain S1 grew more vigorously than strain S1 . Almost no regulation of CrgA protein was observed to 1 gene and . Therefore, CrgA acted as a negative regulator for the medium-to-long-chain -alkane utilization in SJTD-1. The work will promote the regulation mechanism study of -alkane degradation in bacteria and help the bioremediation method development for petroleum pollution.
细菌中的AlkB单加氧酶负责中链和长链烷烃的羟基化。在本研究中,SJTD-1的一种LysR家族成员CrgA蛋白被证明通过直接结合AlkB2单加氧酶基因的上游来调节该酶以及中长链烷烃(C-C)的降解。在AlkB2基因的启动子区域发现了两个CrgA结合的特定位点,并且不完全镜像重复(IIR)结构被证明对CrgA的识别和结合至关重要。十六烷基辅酶A和十八烷基辅酶A可以有效释放CrgA的结合并启动AlkB2基因的转录,这意味着代谢中间产物的正调控。在存在中长链烷烃(C-C)的情况下,缺失CrgA基因可以显著增强AlkB2单加氧酶的转录和表达;在正十八烷培养中,S1菌株比S1菌株生长得更旺盛。几乎未观察到CrgA蛋白对AlkB1基因和AlkB3基因的调控作用。因此,CrgA作为SJTD-1中中长链烷烃利用的负调控因子。这项工作将促进细菌中烷烃降解调控机制的研究,并有助于石油污染生物修复方法的开发。