Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1

doi:10.7717/peerj.6951

IR > 石油化工学院

	Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1
	Luo, Dan 1; Chen, Jixiang1 ; Xie, Gang1 ; Yue, Liang 1; Wang, Yonggang2
	2019-05-21
发表期刊	PEERJ
ISSN	2167-8359
卷号	7
摘要	Resuscitation-promoting factors (Rpf) are a class of muralytic enzymes, which participate in recovery of dormant cells and promoting bacteria growth in poor media. In the present study the expression vector of the rpf-1 gene from an oil-degrading bacterium Rhodococcus erythropolis KB1 was constructed and expressed in Escherichia coli. The expressed protein was purified by Ni2+-affinity chromatography, and showed muralytic activity when measured with 4-methylumbelliferyl-beta-D-N,N',N ''-triacetyl chitotrioside as substrate. Addition of purified Rpf-1 to R. erythropolis culture efficiently improved bacterial cell growth. The purified protein also increased resuscitation of viable but nonculturable cells of R. erythropolis to culturable state. The conserved amino acid residues including Asp(45), Glu(51), Cys(50), Thr(60), Gln(69), Thr(74), Trp(75) and Cys(114) of the Rpf-1 were replaced with different amino acids. The mutant proteins were also expressed and purified with Ni2+-affinity chromatography. The muralytic activities of the mutant proteins decreased to different extents when compared with that of the wild type Rpf-1. Gln(69) was found to play the most important role in the enzyme activity, substitution of Gln(69) with lysine (Q69K) resulted in the greatest decrease of muralytic activity. The other amino acid residues such as Asp(45), Glu(51), Cys(50 )and Cys(114) were also found to be very important in maintaining muralytic activity and biological function of the Rpf-1. Our results indicated that Rpf- 1 from R. erythropolis showed muralytic activities and weak protease activity, but the muralytic activity was responsible for its growth promotion and resuscitation activity.
关键词	Resuscitation promoting factor Rhodococcus erythropolis VBNC Muralytic activity Resuscitation
DOI	10.7717/peerj.6951
收录类别	SCIE
语种	英语
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000468380600003
出版者	PEERJ INC
引用统计	被引频次：7[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://ir.lut.edu.cn/handle/2XXMBERH/155344
专题	石油化工学院生命科学与工程学院
通讯作者	Chen, Jixiang
作者单位	1.Lanzhou Univ Technol, Sch Petrochem Engn, Lanzhou, Gansu, Peoples R China; 2.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou, Gansu, Peoples R China
第一作者单位	兰州理工大学
通讯作者单位	兰州理工大学
第一作者的第一单位	兰州理工大学
推荐引用方式 GB/T 7714	Luo, Dan,Chen, Jixiang,Xie, Gang,et al. Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1[J]. PEERJ,2019,7.
APA	Luo, Dan,Chen, Jixiang,Xie, Gang,Yue, Liang,&Wang, Yonggang.(2019).Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1.PEERJ,7.
MLA	Luo, Dan,et al."Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1".PEERJ 7(2019).