Here, we report on the biochemical characterization of a new glycosylated bacteriocin (glycocin), ASM1, produced by Lactobacillus plantarum A-1 and analysis of the A-1 bacteriocinogenic genes. ASM1 is 43 amino acids in length with Ser18-O- and Cys43-S-linked N-acetylglucosamine moieties that are essential for its inhibitory activity. Its only close homologue, glycocin F (GccF), has five amino acid substitutions all residing in the flexible C-terminal 'tail' and a lower IC50 (0.9 nm) compared to that of ASM1 (1.5 nm). Asm/gcc genes share the same organization (asmH← →asmABCDE→F), and the asm genes reside on an 11 905-bp plasmid dedicated to ASM1 production. The A-1 genome also harbors a gene encoding a 'rare' bactofencin-type bacteriocin. As more examples of prokaryote S-GlcNAcylation are discovered, the functions of this modification may be understood.

Department of Food and Nutritional Sciences Ochanomizu University Tokyo Japan

Institute of Microbiology v v i Academy of Sciences of the Czech Republic Prague 4 Czech Republic

Maurice Wilkins Centre for Molecular Biodiscovery University of Auckland New Zealand

School of Fundamental Sciences Massey University Palmerston North New Zealand

See more in PubMed

Arnison PG, Bibb MJ, Bierbaum G, Bowers AA, Bugni TS, Bulaj G, Camarero JA, Campopiano DJ, Challis GL, Clardy J et al. (2013) Ribosomally synthesized and post-translationally modified peptide natural products: overview and recommendations for a universal nomenclature. Nat Prod Rep 30, 108-160.

Wang H and van der Donk WA (2011) Substrate selectivity of the sublancin S-glycosyltransferase. J Am Chem Soc 133, 16394-16397.

Biswas S, Garcia De Gonzalo CV, Repka LM and van der Donk WA (2017) Structure-activity relationships of the S-linked glycocin sublancin. ACS Chem Biol 12, 2965-2969.

Ren HQ, Biswas S, Ho S, van der Donk WA and Zhao HM (2018) Rapid discovery of glycocins through pathway refactoring in Escherichia coli. ACS Chem Biol 13, 2966-2972.

Oman TJ, Boettcher JM, Wang H, Okalibe XN and van der Donk WA (2011) Sublancin is not a lantibiotic but an S-linked glycopeptide. Nat Chem Biol 7, 78-80.

Kaunietis A, Buivydas A, Citavicius DJ and Kuipers OP (2019) Heterologous biosynthesis and characterization of a glycocin from a thermophilic bacterium. Nat Commun 10, 1115.

Wang H, Oman TJ, Zhang R, Garcia De Gonzalo CV, Zhang Q and van der Donk WA (2014) The glycosyltransferase involved in thurandacin biosynthesis catalyzes both O- and S-glycosylation. J Am Chem Soc 136, 84-87.

Stepper J, Shastri S, Loo TS, Preston JC, Novak P, Man P, Moore CH, Havlicek V, Patchett ML and Norris GE (2011) Cysteine S-glycosylation, a new post-translational modification found in glycopeptide bacteriocins. FEBS Lett 585, 645-650.

Izquierdo E, Wagner C, Marchioni E, Aoude-Werner D and Ennahar S (2009) Enterocin 96, a novel class II bacteriocin produced by Enterococcus faecalis WHE 96. isolated from munster cheese. Appl Environ Microbiol 75, 4273-4276.

Nagar R and Rao A (2017) An iterative glycosyltransferase EntS catalyzes transfer and extension of O- and S-linked monosaccharide in enterocin 96. Glycobiology 27, 766-776.

Maky MA, Ishibashi N, Zendo T, Perez RH, Doud JR, Karmi M and Sonomoto K (2015) Enterocin F4-9, a novel O-linked glycosylated bacteriocin. Appl Environ Microbiol 81, 4819-4826.

Varki A and Lowe JB (2009) Biological roles of glycans. In Essentials of Glycobiology (Varki A, Cummings RD, Esko JD, Freeze HH, Stanley P, Bertozzi CR, Hart GW and Etzler ME, eds), pp. 75-88. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

Liang CF, Yan MC, Chang TC and Lin CC (2009) Synthesis of S-linked alpha(2->9) octasialic acid via exclusive alpha S-glycosidic bond formation. J Am Chem Soc 131, 3138-3139.

Zhu X, Haag T and Schmidt RR (2004) Synthesis of an S-linked glycopeptide analog derived from human Tamm-Horsfall glycoprotein. Org Biomol Chem 2, 31-33.

Hata T, Tanaka R and Ohmomo S (2010) Isolation and characterization of plantaricin ASM1: a new bacteriocin produced by Lactobacillus plantarum A-1. Int J Food Microbiol 137, 94-99.

Trmcic A, Monnet C, Rogelj I and Bogovic Matijasic B (2011) Expression of nisin genes in cheese - a quantitative real-time polymerase chain reaction approach. J Dairy Sci 94, 77-85.

Riley MA and Wertz JE (2002) Bacteriocins: evolution, ecology, and application. Annu Rev Microbiol 56, 117-137.

Klaenhammer TR (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev 12, 39-85.

Cotter PD, Hill C and Ross RP (2005) Bacteriocins: developing innate immunity for food. Nat Rev Microbiol 3, 777-788.

Ahn S, Stepper J, Loo TS, Bisset SW, Patchett ML and Norris GE (2018) Expression of Lactobacillus plantarum KW30 gcc genes correlates with the production of glycocin F in late log phase. FEMS Microbiol Lett 365, fny261.

Kelly WJ, Asmundson RV and Huang CM (1996) Characterization of plantaricin KW30, a bacteriocin produced by Lactobacillus plantarum. J Appl Bacteriol 81, 657-662.

Wells L, Vosseller K, Cole RN, Cronshaw JM, Matunis MJ and Hart GW (2002) Mapping sites of O-GlcNAc modification using affinity tags for serine and threonine post-translational modifications. Mol Cell Proteomics 1, 791-804.

Mikes L and Man P (2003) Purification and characterization of a saccharide-binding protein from penetration glands of Diplostomum pseudospathaceum - a bifunctional molecule with cysteine protease activity. Parasitology 127, 69-77.

O'Shea EF, O'Connor PM, O'Sullivan O, Cotter PD, Ross RP and Hill C (2013) Bactofencin A, a new type of cationic bacteriocin with unusual immunity. MBio 4, e00498-13.

Norris GE and Patchett ML (2014) Glycosylated ribosomally-synthesised peptide toxins: discovery, characterisation and applications. Chapter 14. In Natural Products Analysis: Instrumentation, Methods, and Applications (Havlicek V and Spizek J, eds), pp. 507-543. Wiley, Hoboken, NJ.

Fonseca MM, Harris DJ and Posada D (2014) Origin and length distribution of unidirectional prokaryotic overlapping genes. G3 (Bethesda) 4, 19-27.

Rodriguez-Beltran J, Hernandez-Beltran JCR, DelaFuente J, Escudero JA, Fuentes-Hernandez A, MacLean RC, Peña-Miller R and San Millan A (2018) Multicopy plasmids allow bacteria to escape from fitness trade-offs during evolutionary innovation. Nat Ecol Evol 2, 873-881.

Straume D, Johansen RF, Bjørås M, Nes IF and Diep DB (2009) DNA binding kinetics of two response regulators, PlnC and PlnD, from the bacteriocin regulon of Lactobacillus plantarum C11. BMC Biochem 10, 17.

Zou Z, Qin H, Brenner AE, Raghavan R, Millar JA, Gu Q, Xie Z, Kreth J and Merritt J (2018) LytTR regulatory systems: a potential new class of prokaryotic sensory system. PLoS Genet 14, e1007709.

Norris GE and Patchett ML (2016) The glycocins: in a class of their own. Curr Opin Struct Biol 40, 112-119.

O’ Connor PM, O’ Shea EF, Cotter PD, Hill C and Ross RP (2018) The potency of the broad spectrum bacteriocin, bactofencin A, against staphylococci is highly dependent on primary structure, N-terminal charge and disulphide formation. Sci Rep 8, 11833.

Garcia De Gonzalo CV, Denham EL, Mars RAT, Stulke J, van der Donk WA and van Dijlc JM (2015) The phosphoenolpyruvate: sugar phosphotransferase system is involved in sensitivity to the glucosylated bacteriocin sublancin. Antimicrob Agents Chemother 59, 6844-6854.

Falconer S, Czarny TL and Brown ED (2011) Antibiotics as probes of biological complexity. Nat Chem Biol 7, 415-423.

See more in PubMed

GENBANK
KU896918, AB474371, GU552553, NZ_CP037439, AF304384, CP019716