Cargando…

Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus

Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complet...

Descripción completa

Detalles Bibliográficos
Autores principales: Hong, Sung Wook, Kim, Jong-Hui, Cha, Hyun A, Chung, Kun Sub, Bae, Hyo Ju, Park, Won Seo, Ham, Jun-Sang, Park, Beom-Young, Oh, Mi-Hwa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society for Microbiology and Biotechnology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9720079/
https://www.ncbi.nlm.nih.gov/pubmed/36310361
http://dx.doi.org/10.4014/jmb.2208.08006
_version_ 1784843471030321152
author Hong, Sung Wook
Kim, Jong-Hui
Cha, Hyun A
Chung, Kun Sub
Bae, Hyo Ju
Park, Won Seo
Ham, Jun-Sang
Park, Beom-Young
Oh, Mi-Hwa
author_facet Hong, Sung Wook
Kim, Jong-Hui
Cha, Hyun A
Chung, Kun Sub
Bae, Hyo Ju
Park, Won Seo
Ham, Jun-Sang
Park, Beom-Young
Oh, Mi-Hwa
author_sort Hong, Sung Wook
collection PubMed
description Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0–70°C and pH 2–10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15.
format Online
Article
Text
id pubmed-9720079
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher The Korean Society for Microbiology and Biotechnology
record_format MEDLINE/PubMed
spelling pubmed-97200792022-12-13 Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus Hong, Sung Wook Kim, Jong-Hui Cha, Hyun A Chung, Kun Sub Bae, Hyo Ju Park, Won Seo Ham, Jun-Sang Park, Beom-Young Oh, Mi-Hwa J Microbiol Biotechnol Research article Natural antimicrobial substances are needed as alternatives to synthetic antimicrobials to protect against foodborne pathogens. In this study, a bacteriocin-producing bacterium, Bacillus subtilis HD15, was isolated from doenjang, a traditional Korean fermented soybean paste. We sequenced the complete genome of B. subtilis HD15. This genome size was 4,173,431 bp with a G + C content of of 43.58%, 4,305 genes, and 4,222 protein-coding genes with predicted functions, including a subtilosin A gene cluster. The bacteriocin was purified by ammonium sulfate precipitation, Diethylaminoethanol-Sepharose chromatography, and Sephacryl gel filtration, with 12.4-fold purification and 26.2% yield, respectively. The purified protein had a molecular weight of 3.6 kDa. The N-terminal amino acid sequence showed the highest similarity to Bacillus subtilis 168 subtilosin A (78%) but only 68% similarity to B. tequilensis subtilosin proteins, indicating that the antimicrobial substance isolated from B. subtilis HD15 is a novel bacteriocin related to subtilosin A. The purified protein from B. subtilis HD15 exhibited high antimicrobial activity against Listeria monocytogenes and Bacillus cereus. It showed stable activity in the range 0–70°C and pH 2–10 and was completely inhibited by protease, proteinase K, and pronase E treatment, suggesting that it is a proteinaceous substance. These findings support the potential industrial applications of the novel bacteriocin purified from B. subtilis HD15. The Korean Society for Microbiology and Biotechnology 2022-11-28 2022-10-14 /pmc/articles/PMC9720079/ /pubmed/36310361 http://dx.doi.org/10.4014/jmb.2208.08006 Text en Copyright © 2022 by the authors. Licensee KMB. https://creativecommons.org/licenses/by/4.0/This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research article
Hong, Sung Wook
Kim, Jong-Hui
Cha, Hyun A
Chung, Kun Sub
Bae, Hyo Ju
Park, Won Seo
Ham, Jun-Sang
Park, Beom-Young
Oh, Mi-Hwa
Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title_full Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title_fullStr Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title_full_unstemmed Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title_short Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus
title_sort identification and characterization of a bacteriocin from the newly isolated bacillus subtilis hd15 with inhibitory effects against bacillus cereus
topic Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9720079/
https://www.ncbi.nlm.nih.gov/pubmed/36310361
http://dx.doi.org/10.4014/jmb.2208.08006
work_keys_str_mv AT hongsungwook identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT kimjonghui identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT chahyuna identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT chungkunsub identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT baehyoju identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT parkwonseo identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT hamjunsang identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT parkbeomyoung identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus
AT ohmihwa identificationandcharacterizationofabacteriocinfromthenewlyisolatedbacillussubtilishd15withinhibitoryeffectsagainstbacilluscereus