Identification and three‐dimensional structure of carnobacteriocin XY, a class IIb bacteriocin produced by <i>Carnobacteria</i>

Acedo, Jeella Z.; Towle, Kaitlyn M.; Lohans, Christopher T.; Miskolzie, Mark; McKay, Ryan; Doerksen, Thomas; Vederas, John C.; Martin‐Visscher, Leah A.

doi:10.1002/1873-3468.12648

Cited by 22 publications

(23 citation statements)

References 44 publications

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“…The carnobacteriocin BM1 BGC [ 56 ] was found in the C. gallinarum MT44 genome and in 85% of the C. maltaromaticum genomes. Other described UBs were exclusively found in genomes of C. maltaromaticum species—the piscicolin 126 BGC [ 57 ] in 15% of the genomes, and the carnobacteriocin B2 and X/Y BGCs [ 56 , 58 ] in 38% of the genomes. The divergicin A BGC [ 59 ] was identified in the C. divergens B7 and C8 strains and the divercin V41 BGC [ 48 ] only in the V41 strain.…”

Section: Resultsmentioning

confidence: 99%

New Insight into Antimicrobial Compounds from Food and Marine-Sourced Carnobacterium Species through Phenotype and Genome Analyses

et al. 2020

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Carnobacterium maltaromaticum and Carnobacterium divergens, isolated from food products, are lactic acid bacteria known to produce active and efficient bacteriocins. Other species, particularly those originating from marine sources, are less studied. The aim of the study is to select promising strains with antimicrobial potential by combining genomic and phenotypic approaches on large datasets comprising 12 Carnobacterium species. The biosynthetic gene cluster (BGCs) diversity of 39 publicly available Carnobacterium spp. genomes revealed 67 BGCs, distributed according to the species and ecological niches. From zero to six BGCs were predicted per strain and classified into four classes: terpene, NRPS (non-ribosomal peptide synthetase), NRPS-PKS (hybrid non-ribosomal peptide synthetase-polyketide synthase), RiPP (ribosomally synthesized and post-translationally modified peptide). In parallel, the antimicrobial activity of 260 strains from seafood products was evaluated. Among the 60% of active strains, three genomes were sequenced and submitted to a dereplication process. C. inhibens MIP2551 produced a high amountof H2O2, probably thanks to the presence of four oxidase-encoding genes. C. maltaromaticum EBP3019 and SF668 strains were highly efficient against Listeria monocytogenes. A new extracellular 16 kDa unmodified bacteriocin in the EBP3019 strain and five different bacteriocins in SF668 were highlighted. In this study, the overview of antimicrobial BGC and inhibitory activities of Carnobacterium spp. allowed the prediction of potential innovative natural products that could be relevant for biotechnological applications.

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Section: Resultsmentioning

confidence: 99%

New Insight into Antimicrobial Compounds from Food and Marine-Sourced Carnobacterium Species through Phenotype and Genome Analyses

et al. 2020

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“…gasseri K7, was reported 23 , indicating that the structure of Gas K7_Acp may be different from inactive GatX in this study, owing to test solutions and/or expression hosts used, in spite of the same indicator strain and amino acid sequences. Each component peptide (CbnX and CbnY) of class IIb bacteriocin carnobacteriocin XY presented high α-helix content in trifluoroethanol solution, while CbnXY did not show remarkable second structures in aqueous conditions 24 . Furthermore, our previous study for isolation and purification of GatA and GatX from the culture supernatant of Lb.…”

Section: Discussionmentioning

confidence: 95%

Homologous expression and characterization of gassericin T and gassericin S, a novel class IIb bacteriocin produced byLactobacillus gasseriLA327

Genki

Tanaka

Harada

et al. 2018

Preprint

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23Lactobacillus gasseri LA327 isolated from the large intestine tissue in humans is a 24 bacteriocinogenic strain and is predicted to produce two kinds of class IIb bacteriocins, i.e. 25 gassericin T (GT) and acidocin LF221A (Acd LF221A). In this study, DNA sequencing of the 26 genes for GT and Acd LF221A on Lb. gasseri LA327 revealed that the amino acid sequences for 27 GT completely corresponded with those of gat except for GatK (histidine kinase). However, 28 those for the Acd LF221A had analogues which differed in at least one amino acid residue to be a 29 putative class IIb bacteriocin designated as gassericin S (GS). By deletion test of GT structural 30 genes (gatAX), the LA327 strain retained the bacteriocin activity, and the LA327 mutant strain 31 lacking the ABC-type transporter gene (gatT) completely lost the bacteriocin activity. This 32 indicates that LA327 strain is a GS producer, and GS production is performed via gat with the 33 inclusion of gatT. Homologous expression using deletion mutants for GS and GT containing 34 each single peptide elucidated that GS (GasAX) and GT (GatAX) showed synergistic activity as 35 class IIb bacteriocins, respectively, and no synergistic activity was observed between each 36 peptide of GS and GT. The molecular mass of GS was estimated to be theoretical ca. 5,400 Da 37 by in situ activity assay after SDS-PAGE, clarifying that GS was actually expressed as an active 38 class IIb bacteriocin. Furthermore, stability of GS expressed against pH, heat and protease was 39 determined. 40 41 Importance 42We determined the complete DNA sequence for GS, a novel class IIb bacteriocin of Lb. 43 gasseri, and succeeded to express GS as active bacteriocins. Our results clarified the interaction 44 3 of each class IIb component peptide for GT in addition to GS via construction of homologous 45 mutants which were not dependent on the purification. These data may demonstrate the 46 characteristics of class IIb bacteriocins for Lb. gasseri. 47 48 Lactic acid bacteria (LAB), one of the potent candidates which are generally recognized 50 as safe (GRAS) microorganisms, have a long history of consumption by humans, and inhabiting 51 various ecosystems, including gastrointestinal (GI) tracts of humans and animals, and other 52 environments (for instance, vegetables, milks, and meats). In addition, lactobacilli are recognized 53 as important members of beneficial GI microbiota of humans and animals, along with 54 bifidobacteria 1 . Especially, Lactobacillus gasseri is one of the predominant species in 55 lactobacilli of human small intestines, and has been isolated not only from the GI tracts and the 56 feces, but also the oral and the vaginal cavities and mammary areola, and is considered one of 57 the representative probiotics 2-5 . 58 Many researchers have reported the beneficial effects of Lb. gasseri on the hosts, 59including the immunoregulation, the alleviation of allergic symptoms, the prevention of bacterial 60 and viral infections, the antitumor effect, and the inhibition of lip...

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“…bulgaricus LMG 6901 T (JCM 1002 T ) of the second peptide for gassericin K7B, 100% homologous to the GT second peptide, was reported (23), indicating that the structure of the gassericin K7B second peptide (active) may be different from that of the GT second peptide (inactive) in this study, owing to the test solutions and/or expression hosts used, in spite of the same indicator strain and amino acid sequences being used. Component peptides of the class IIb bacteriocin carnobacteriocin XY showed a high ␣-helix content in trifluoroethanol solution, while neither peptide showed remarkable secondary structure in aqueous conditions (30). Furthermore, our previous attempts to isolate and purify individual GT peptides from the CFS of L. gasseri LA158 failed because of polymerization of the peptides in 60% 2-propanol at the final step for high-pressure liquid chromatography (HPLC) purification (20).…”

Section: Discussionmentioning

confidence: 99%

Homologous Expression and Characterization of Gassericin T and Gassericin S, a Novel Class IIb Bacteriocin Produced by Lactobacillus gasseri LA327

Genki

Tanaka

Harada

et al. 2019

Appl Environ Microbiol

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Lactobacillus gasseri LA327, isolated from the large intestine tissue in humans, is a bacteriocinogenic strain with two kinds of class IIb bacteriocin structural genes, i.e., those for gassericin T (GT) and acidocin LF221A (Acd LF221A). In this study, DNA sequencing of the genes for GT and Acd LF221A from L. gasseri LA327 revealed that the amino acid sequences for GT corresponded with those for GT genes, except for GatK (histidine kinase). However, Acd LF221A genes had analogues which differed in at least one amino acid residue, to encode a class IIb bacteriocin designated gassericin S (GS). The LA327 strain retained antimicrobial activity after the deletion of the GT structural genes (gatAX); however, both GS and GT activities were lost by deletion of the putative ABC transporter gene (gatT). This indicates that the LA327 strain produces GS and GT and that GS secretion is performed via GT genes with the inclusion of gatT. Homologous expression using deletion mutants of GS and GT, each containing a single peptide, elucidated that GS (GasAX) and GT (GatAX) showed synergistic activity as class IIb bacteriocins and that no synergistic activity was observed between GS and GT peptides. The molecular mass of GS was estimated to be theoretical ca. 5,400 Da by in situ activity assay after SDS-PAGE, clarifying that GS was actually expressed as an active class IIb bacteriocin. Furthermore, the stability of expressed GS to pH, heat, and protease was determined. IMPORTANCE Bacteriocins are regarded as potential alternatives for antibiotics in the absence of highly resistant bacteria. In particular, two-peptide (class IIb) bacteriocins exhibit the maximum activity through the synergy of two components, and their antimicrobial spectra are known to be relatively wide. However, there are few reports of synergistic activity of class IIb bacteriocins determined by isolation and purification of individual peptides. Our results clarified the interaction of each class IIb component peptide for GT and GS via the construction of homologous mutants, which were not dependent on the purification. These data may contribute to understanding the mechanisms of action by which class IIb bacteriocins exhibit wide antibacterial spectra.

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Identification and three‐dimensional structure of carnobacteriocin XY, a class IIb bacteriocin produced by Carnobacteria

Cited by 22 publications

References 44 publications

New Insight into Antimicrobial Compounds from Food and Marine-Sourced Carnobacterium Species through Phenotype and Genome Analyses

New Insight into Antimicrobial Compounds from Food and Marine-Sourced Carnobacterium Species through Phenotype and Genome Analyses

Homologous expression and characterization of gassericin T and gassericin S, a novel class IIb bacteriocin produced byLactobacillus gasseriLA327

Homologous Expression and Characterization of Gassericin T and Gassericin S, a Novel Class IIb Bacteriocin Produced by Lactobacillus gasseri LA327

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