TY - JOUR
T1 - Rapid Discovery of Glycocins through Pathway Refactoring in Escherichia coli
AU - Ren, Hengqian
AU - Biswas, Subhanip
AU - Ho, Sherri
AU - Van Der Donk, Wilfred A.
AU - Zhao, Huimin
N1 - Funding Information:
This work was supported by the National Institutes of Health (GM077596) (H.Z. and W.A.V.) and the Howard Hughes Medical Institute (W.A.V.). We thank A. Ulanov in the Roy J. Carver Biotechnology Center at the University of Illinois at Urbana−Champaign for assistance with the GC-MS analysis. We thank P. Yau in the Roy J. Carver Biotechnology Center at the University of Illinois at Urbana−Champaign for assistance with the MS/MS analysis.
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/10/19
Y1 - 2018/10/19
N2 - Glycocins (glycosylated bacteriocins) are a family of ribosomally synthesized and post-translationally modified peptides with antimicrobial activities against pathogens of interest, including methicillin-resistant Staphylococcus aureus, representing a promising source of new antibiotics. Glycocins are still largely underexplored, and thus far, only six glycocins are known. Here, we used genome mining to identify 50 putative glycocin biosynthetic gene clusters and then chose six of them with distinct features for further investigation. Through two rounds of plug-and-play pathway refactoring and expression in Escherichia coli BL21(DE3), four systems produced novel glycocins. Further structural characterization revealed that one of them, which belongs to the enterocin 96-type glycocins, was diglucosylated on a single serine. The other three compounds belong to the SunA/ThuA-type glycocins and exhibit a antimicrobial spectrum narrower than that of sublancin, the best characterized member in this group, even though they share a similar disulfide topology and glycosylation. Further evaluation of their bioactivities with free glucose at high concentrations suggested that their antimicrobial mechanisms might be both glycocin- and species-specific. These glycocins with distinct features significantly broaden our knowledge and may lead to the discovery of new classes of antibiotics.
AB - Glycocins (glycosylated bacteriocins) are a family of ribosomally synthesized and post-translationally modified peptides with antimicrobial activities against pathogens of interest, including methicillin-resistant Staphylococcus aureus, representing a promising source of new antibiotics. Glycocins are still largely underexplored, and thus far, only six glycocins are known. Here, we used genome mining to identify 50 putative glycocin biosynthetic gene clusters and then chose six of them with distinct features for further investigation. Through two rounds of plug-and-play pathway refactoring and expression in Escherichia coli BL21(DE3), four systems produced novel glycocins. Further structural characterization revealed that one of them, which belongs to the enterocin 96-type glycocins, was diglucosylated on a single serine. The other three compounds belong to the SunA/ThuA-type glycocins and exhibit a antimicrobial spectrum narrower than that of sublancin, the best characterized member in this group, even though they share a similar disulfide topology and glycosylation. Further evaluation of their bioactivities with free glucose at high concentrations suggested that their antimicrobial mechanisms might be both glycocin- and species-specific. These glycocins with distinct features significantly broaden our knowledge and may lead to the discovery of new classes of antibiotics.
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U2 - 10.1021/acschembio.8b00599
DO - 10.1021/acschembio.8b00599
M3 - Article
C2 - 30183259
AN - SCOPUS:85053727239
SN - 1554-8929
VL - 13
SP - 2966
EP - 2972
JO - ACS chemical biology
JF - ACS chemical biology
IS - 10
ER -