Functional genetic analysis of the GarML gene cluster in Lactococcus garvieae DCC43 gives new insights into circular bacteriocin biosynthesis

Journal of Bacteriology
Christina GabrielsenDzung B Diep

Abstract

Garvicin ML (GarML) is a circular bacteriocin produced by Lactococcus garvieae DCC43. The recently published draft genome of this strain allowed determination of the genetic background for bacteriocin production. Bioinformatic analysis identified a gene cluster consisting of nine open reading frames likely involved in the production of and immunity to GarML. The garA gene encodes the bacteriocin precursor, garX a large transmembrane protein, garBCDE a putative immunity protein (garB) followed by an ATPase and two transmembrane proteins, and garFGH a putative ABC transporter complex. Functional genetic analysis revealed that deletion of garFGH had no effect on sensitivity to or production of GarML. In contrast, deletion of garBCDE or inactivation of garX resulted in high-level sensitivity to GarML and completely abolished production of active bacteriocin. Mass spectrometry of culture supernatants revealed that wild-type cultures contained the mature circular form as well as the linear forms of the bacteriocin, both with and without the three-amino-acid leader sequence, while bacteriocin-negative mutants contained only the linear forms. These results indicate that cleavage of the leader peptide precedes circularization and is lik...Continue Reading

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Citations

Feb 11, 2016·Applied Microbiology and Biotechnology·Patricia Alvarez-SieiroOscar P Kuipers
Jun 19, 2016·Applied and Environmental Microbiology·Kirill V OvchinnikovDzung B Diep
Aug 31, 2014·Applied and Environmental Microbiology·Christina GabrielsenDzung B Diep
Sep 17, 2020·Natural Product Reports·Manuel Montalbán-LópezWilfred A van der Donk
Sep 21, 2018·Frontiers in Microbiology·Rodney H PerezKenji Sonomoto
Dec 29, 2020·Critical Reviews in Food Science and Nutrition·Silpa S, Rupachandra S

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