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

doi:10.7717/peerj.6951

CORC > 兰州理工大学 > 兰州理工大学 > 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, Jixiang 1; Xie, Gang 1; Yue, Liang 1; Wang, Yonggang 2
刊名	PEERJ
	2019-05-21
卷号	7
关键词	Resuscitation promoting factor Rhodococcus erythropolis VBNC Muralytic activity Resuscitation
ISSN号	2167-8359
DOI	10.7717/peerj.6951
英文摘要	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.
WOS研究方向	Science & Technology - Other Topics
语种	英语
出版者	PEERJ INC
WOS记录号	WOS:000468380600003
内容类型	期刊论文
源URL	[http://ir.lut.edu.cn/handle/2XXMBERH/155344]
专题	石油化工学院生命科学与工程学院
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
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).