Enzyme Characterization and Biological Activities of a Resuscitation Promoting Factor from an Oil Degrading Bacterium KB1

Overview

Journal PeerJ

Specialties Biology
Environmental Health
General Medicine

Date 2019 Jun 1

PMID 31149404

Citations 4

Authors

Dan Luo

Jixiang Chen

Gang Xie

Liang Yue

Yonggang Wang

Affiliations

Soon will be listed here.

Abstract

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 gene from an oil-degrading bacterium KB1 was constructed and expressed in The expressed protein was purified by Ni-affinity chromatography, and showed muralytic activity when measured with 4-methylumbelliferyl-β-D-N,N',N″-triacetyl chitotrioside as substrate. Addition of purified Rpf-1 to culture efficiently improved bacterial cell growth. The purified protein also increased resuscitation of viable but nonculturable cells of to culturable state. The conserved amino acid residues including Asp, Glu, Cys, Thr, Gln, Thr, Trp and Cys of the Rpf-1 were replaced with different amino acids. The mutant proteins were also expressed and purified with Ni-affinity chromatography. The muralytic activities of the mutant proteins decreased to different extents when compared with that of the wild type Rpf-1. Gln was found to play the most important role in the enzyme activity, substitution of Gln with lysine (Q69K) resulted in the greatest decrease of muralytic activity. The other amino acid residues such as Asp, Glu, Cys and Cys 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 showed muralytic activities and weak protease activity, but the muralytic activity was responsible for its growth promotion and resuscitation activity.

Citing Articles

The DUF348 domains of resuscitation promoting factor 2 play important roles in the enzymatic and biological activities in KB1.

Fu J, Chen J, Wang Y, Luo D, Wang T, Zhang Q PeerJ. 2024; 12:e18561.

PMID: 39583115 PMC: 11583912. DOI: 10.7717/peerj.18561.

Resuscitation Promotion Factor: A Pronounced Bacterial Cytokine in Propelling Bacterial Resuscitation.

Li X, Ren Q, Sun Z, Wu Y, Pan H Microorganisms. 2024; 12(8).

PMID: 39203370 PMC: 11356341. DOI: 10.3390/microorganisms12081528.

Viable but nonculturable bacteria and their resuscitation: implications for cultivating uncultured marine microorganisms.

Zhang X, Ahmad W, Zhu X, Chen J, Austin B Mar Life Sci Technol. 2023; 3(2):189-203.

PMID: 37073345 PMC: 10077291. DOI: 10.1007/s42995-020-00041-3.

Enzymatic properties and biological activity of resuscitation-promoting factor B of sp. (GX12401).

Gong X, Lu H, Wu J, Zhou Y, Yang L, Wang Y Front Microbiol. 2022; 13:965843.

PMID: 36274735 PMC: 9580463. DOI: 10.3389/fmicb.2022.965843.

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