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Characterisation of the Brochothrix thermosphacta sortase A enzyme

Citation

Stanborough, T and Suryadinata, R and Fegan, N and Powell, SM and Tamplin, M and Nuttall, SD and Scott Chandry, P, Characterisation of the Brochothrix thermosphacta sortase A enzyme, FEMS Microbiology Letters, 365, (17) pp. 1-9. ISSN 0378-1097 (2018) [Refereed Article]

Copyright Statement

FEMS 2018.

DOI: doi:10.1093/femsle/fny184

Abstract

Gram-positive bacteria utilise class A sortases to coat the surface of their cells with a diversity of proteins that facilitate interactions with their environment and play fundamental roles in cell physiology and virulence. A putative sortase A gene was identified in the genome of the poorly studied meat spoilage bacterium Brochothrix thermosphacta. To understand how this bacterium mediates interactions with its environment, an N-terminal truncated, His-tagged variant of this protein (His6-BtSrtA) was expressed and purified. Catalytic activity of recombinant His6-BtSrtA was investigated, including sorting motif recognition of target proteins and bioconjugation activity. Further, the B. thermosphacta genome was examined for the presence of sortase A (SrtA) protein substrates. His6-BtSrtA readily formed intermediate complexes with LPXTG-tagged proteins. Although the reaction was inefficient, nucleophilic attack of the resultant thioacyl intermediates by tri-glycine was observed. Genome examination identified 11 potential SrtA substrates, two of which contained protein domains associated with adherence of pathogens to host extracellular matrix proteins and cells, suggesting the B. thermosphacta SrtA may be indirectly involved in its attachment to meat surfaces. Thus, further work in this area could provide crucial insight into molecular mechanisms involved in the colonisation of meat by B. thermosphacta.

Item Details

Item Type:Refereed Article
Keywords:sortase; SrtA; Brochothrix thermosphacta; meat spoilage; LPXTG; bioconjugation
Research Division:Biological Sciences
Research Group:Microbiology
Research Field:Bacteriology
Objective Division:Health
Objective Group:Public health (excl. specific population health)
Objective Field:Food safety
UTAS Author:Powell, SM (Ms Sophie Powell)
UTAS Author:Tamplin, M (Professor Mark Tamplin)
ID Code:152682
Year Published:2018
Web of Science® Times Cited:2
Deposited By:Research Division
Deposited On:2022-08-23
Last Modified:2022-09-08
Downloads:0

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