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The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae

Vibrio cholerae can survive cold stress by entering into a viable but non-culturable (VBNC) state, and resuscitation can be induced either by temperature upshift only or the addition of an anti-dormancy stimulant such as resuscitation-promoting factors (Rpfs) at suitable temperature. In this study,...

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Autores principales: Debnath, Anusuya, Miyoshi, Shin-ichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707003/
https://www.ncbi.nlm.nih.gov/pubmed/34946219
http://dx.doi.org/10.3390/microorganisms9122618
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author Debnath, Anusuya
Miyoshi, Shin-ichi
author_facet Debnath, Anusuya
Miyoshi, Shin-ichi
author_sort Debnath, Anusuya
collection PubMed
description Vibrio cholerae can survive cold stress by entering into a viable but non-culturable (VBNC) state, and resuscitation can be induced either by temperature upshift only or the addition of an anti-dormancy stimulant such as resuscitation-promoting factors (Rpfs) at suitable temperature. In this study, the role of proteinase K was analyzed as an Rpf in V. cholerae. A VBNC state was induced in V. cholerae AN59 in artificial seawater (ASW) media at 4 °C, and recovery could be achieved in filtered VBNC microcosm, called spent ASW media, merely by a temperature upshift to 37 °C. The resuscitation ability of spent ASW was further enhanced by the addition of proteinase K. The mode of action of proteinase K was investigated by comparing its effect on the growth of the VBNC and culturable state of V. cholerae in ASW and spent ASW media. The presence of proteinase K allowed culturable cells to grow faster in ASW by reducing the generation time. However, this effect of proteinase K was more pronounced in stressed VBNC cells. Moreover, proteinase K-supplemented spent ASW could also accelerate the transition of VBNC into recovered cells followed by rapid growth. Additionally, we found that dead bacterial cells were the substrate on which proteinase K acts to support high growth in spent ASW. So, the conclusion is that the proteinase K could efficiently promote the recovery and growth of dormant VBNC cells at higher temperatures by decreasing the duration of the initial lag phase required for transitioning from the VBNC to recovery state and increasing the growth rate of these recovered cells.
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spelling pubmed-87070032021-12-25 The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae Debnath, Anusuya Miyoshi, Shin-ichi Microorganisms Article Vibrio cholerae can survive cold stress by entering into a viable but non-culturable (VBNC) state, and resuscitation can be induced either by temperature upshift only or the addition of an anti-dormancy stimulant such as resuscitation-promoting factors (Rpfs) at suitable temperature. In this study, the role of proteinase K was analyzed as an Rpf in V. cholerae. A VBNC state was induced in V. cholerae AN59 in artificial seawater (ASW) media at 4 °C, and recovery could be achieved in filtered VBNC microcosm, called spent ASW media, merely by a temperature upshift to 37 °C. The resuscitation ability of spent ASW was further enhanced by the addition of proteinase K. The mode of action of proteinase K was investigated by comparing its effect on the growth of the VBNC and culturable state of V. cholerae in ASW and spent ASW media. The presence of proteinase K allowed culturable cells to grow faster in ASW by reducing the generation time. However, this effect of proteinase K was more pronounced in stressed VBNC cells. Moreover, proteinase K-supplemented spent ASW could also accelerate the transition of VBNC into recovered cells followed by rapid growth. Additionally, we found that dead bacterial cells were the substrate on which proteinase K acts to support high growth in spent ASW. So, the conclusion is that the proteinase K could efficiently promote the recovery and growth of dormant VBNC cells at higher temperatures by decreasing the duration of the initial lag phase required for transitioning from the VBNC to recovery state and increasing the growth rate of these recovered cells. MDPI 2021-12-18 /pmc/articles/PMC8707003/ /pubmed/34946219 http://dx.doi.org/10.3390/microorganisms9122618 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. 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 Article
Debnath, Anusuya
Miyoshi, Shin-ichi
The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title_full The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title_fullStr The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title_full_unstemmed The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title_short The Impact of Protease during Recovery from Viable but Non-Culturable (VBNC) State in Vibrio cholerae
title_sort impact of protease during recovery from viable but non-culturable (vbnc) state in vibrio cholerae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707003/
https://www.ncbi.nlm.nih.gov/pubmed/34946219
http://dx.doi.org/10.3390/microorganisms9122618
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