Characterization of a New M13 Metallopeptidase from Deep-Sea Shewanella sp. E525-6 and Mechanistic Insight into Its Catalysis

Frontiers in Microbiology
Jin-Yu YangXiu-Lan Chen

Abstract

Bacterial extracellular peptidases are important for bacterial nutrition and organic nitrogen degradation in the ocean. While many peptidases of the M13 family from terrestrial animals and bacteria are studied, there has been no report on M13 peptidases from marine bacteria. Here, we characterized an M13 peptidase, PepS, from the deep-sea sedimentary strain Shewanella sp. E525-6, and investigated its substrate specificity and catalytic mechanism. The gene pepS cloned from strain E525-6 contains 2085 bp and encodes an M13 metallopeptidase. PepS was expressed in Escherichia coli and purified. Among the characterized M13 peptidases, PepS shares the highest sequence identity (47%) with Zmp1 from Mycobacterium tuberculosis, indicating that PepS is a new member of the M13 family. PepS had the highest activity at 30°C and pH 8.0. It retained 15% activity at 0°C. Its half life at 40°C was only 4 min. These properties indicate that PepS is a cold-adapted enzyme. The smallest substrate for PepS is pentapeptide, and it is probably unable to cleave peptides of more than 30 residues. PepS prefers to hydrolyze peptide bonds with P1' hydrophobic residues. Structural and mutational analyses suggested that His531, His535 and Glu592 coordinate t...Continue Reading

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Citations

Nov 22, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jie YangXiu-Lan Chen
Jan 8, 2020·International Journal of Systematic and Evolutionary Microbiology·Guillaume TahonAnne Willems

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Datasets Mentioned

BETA
KT591706
KT692986

Methods Mentioned

BETA
PCR
gel filtration
electrophoresis
protein assay
Circular dichroism

Software Mentioned

BLAST
Model
SWISS
PyMOL
ClustalW
ESPript
SignalP
ExPASy

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