Heterologous biosynthesis and characterization of a glycocin from a thermophilic bacterium

Abstract

The genome of the thermophilic bacterium, Aeribacillus pallidus 8, encodes the bacteriocin pallidocin. It belongs to the small class of glycocins and is posttranslationally modified, containing an S -linked glucose on a specific Cys residue. In this study, the pallidocin biosynthetic machinery is cloned and expressed in Escherichia coli to achieve its full biosynthesis and modification. It targets other thermophilic bacteria with potent activity, demonstrated by a low minimum inhibitory concentration (MIC) value. Moreover, the characterized biosynthetic machinery is employed to produce two other glycopeptides Hyp1 and Hyp2. Pallidocin and Hyp1 exhibit antibacterial activity against closely related thermophilic bacteria and some Bacillus sp. strains. Thus, heterologous expression of a glycocin biosynthetic gene cluster including an S -glycosyltransferase provides a good tool for production of hypothetical glycocins encoded by various bacterial genomes and allows rapid in vivo screening.

Biosynthesis and Heterologous Expression of Cacaoidin, the First Member of the Lanthidin Family of RiPPs

“…In addition to cacaoidin, only two glycosylated RiPPs produced by bacteria have been described: the group of glycocins, whose sugar moieties are linked to Cys, Ser or Thr residues by S-glycosyltransferases [ 40 ], and the class III lanthipeptide NAI-112, which carries a 6-deoxyhexose moiety N-linked to a tryptophan residue by the AplG GT [ 41 ]. However, low homologies were found between these enzymes and the GTs present in the cacaoidin cluster.…”

Bacteriocin ASM1 is an O/S‐diglycosylated, plasmid‐encoded homologue of glycocin F

“…The two Enterococcus glycocins are modified by O‐linked sugars only , the two Bacillus and the Aeribacillus glycocins by an S‐linked glucose , and glycocin F is both O‐ and S‐GlcNAcylated . Protein sidechain glycosylation is generally considered to fulfill physicochemical and/or molecular recognition roles .…”

Recent Development of Extremophilic Bacteria and Their Application in Biorefinery

“…The inoculation of Consortium TC-5 in digested sludge was reported to significantly increase the methane production rate by 36.6% under thermophilic conditions, which is conducive to the sustainable development of the industrial economy (Kong, 2018). Most recently, a glycocin biosynthetic gene cluster from a thermophilic Aeribacillus pallidus 8 was expressed heterologously in E. coli as a good tool for the production of hypothetical glycocin, which holds the potential application in biofuel production (Arnoldas Kaunietis et al, 2019). In a recent study, a butanol-producing Clostridium sp.…”