Cargando…

Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis

BACKGROUND: Bacillus subtilis is an important industrial workhorse applied in the production of many different commercially relevant proteins, especially enzymes. Virtually all of these proteins are secreted via the general secretion (Sec) pathway. Studies from different laboratories have demonstrat...

Descripción completa

Detalles Bibliográficos
Autores principales: Neef, Jolanda, Bongiorni, Cristina, Schmidt, Brian, Goosens, Vivianne J., van Dijl, Jan Maarten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7048088/
https://www.ncbi.nlm.nih.gov/pubmed/32111210
http://dx.doi.org/10.1186/s12934-020-01315-2
_version_ 1783502236218818560
author Neef, Jolanda
Bongiorni, Cristina
Schmidt, Brian
Goosens, Vivianne J.
van Dijl, Jan Maarten
author_facet Neef, Jolanda
Bongiorni, Cristina
Schmidt, Brian
Goosens, Vivianne J.
van Dijl, Jan Maarten
author_sort Neef, Jolanda
collection PubMed
description BACKGROUND: Bacillus subtilis is an important industrial workhorse applied in the production of many different commercially relevant proteins, especially enzymes. Virtually all of these proteins are secreted via the general secretion (Sec) pathway. Studies from different laboratories have demonstrated essential or non-essential contributions of various Sec machinery components to protein secretion in B. subtilis. However, a systematic comparison of the impact of each individual Sec machinery component under conditions of high-level protein secretion was so far missing. RESULTS: In the present study, we have compared the contributions of non-essential Sec pathway components and cell envelope-associated proteases on the secretion efficiency of three proteins expressed at high level. This concerned the α-amylases AmyE from B. subtilis and AmyL from Bacillus licheniformis, and the serine protease BPN’ from Bacillus amyloliquefaciens. We compared the secretion capacity of mutant strains in shake flask cultures, and the respective secretion kinetics by pulse-chase labeling experiments. The results show that secDF, secG or rasP mutations severely affect AmyE, AmyL and BPN’ secretion, but the actual effect size depends on the investigated protein. Additionally, the chaperone DnaK is important for BPN’ secretion, while AmyE or AmyL secretion are not affected by a dnaK deletion. Further, we assessed the induction of secretion stress responses in mutant strains by examining AmyE- and AmyL-dependent induction of the quality control proteases HtrA and HtrB. Interestingly, the deletion of certain sip genes revealed a strong differential impact of particular signal peptidases on the magnitude of the secretion stress response. CONCLUSIONS: The results of the present study highlight the importance of SecDF, SecG and RasP for protein secretion and reveal unexpected differences in the induction of the secretion stress response in different mutant strains.
format Online
Article
Text
id pubmed-7048088
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-70480882020-03-05 Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis Neef, Jolanda Bongiorni, Cristina Schmidt, Brian Goosens, Vivianne J. van Dijl, Jan Maarten Microb Cell Fact Research BACKGROUND: Bacillus subtilis is an important industrial workhorse applied in the production of many different commercially relevant proteins, especially enzymes. Virtually all of these proteins are secreted via the general secretion (Sec) pathway. Studies from different laboratories have demonstrated essential or non-essential contributions of various Sec machinery components to protein secretion in B. subtilis. However, a systematic comparison of the impact of each individual Sec machinery component under conditions of high-level protein secretion was so far missing. RESULTS: In the present study, we have compared the contributions of non-essential Sec pathway components and cell envelope-associated proteases on the secretion efficiency of three proteins expressed at high level. This concerned the α-amylases AmyE from B. subtilis and AmyL from Bacillus licheniformis, and the serine protease BPN’ from Bacillus amyloliquefaciens. We compared the secretion capacity of mutant strains in shake flask cultures, and the respective secretion kinetics by pulse-chase labeling experiments. The results show that secDF, secG or rasP mutations severely affect AmyE, AmyL and BPN’ secretion, but the actual effect size depends on the investigated protein. Additionally, the chaperone DnaK is important for BPN’ secretion, while AmyE or AmyL secretion are not affected by a dnaK deletion. Further, we assessed the induction of secretion stress responses in mutant strains by examining AmyE- and AmyL-dependent induction of the quality control proteases HtrA and HtrB. Interestingly, the deletion of certain sip genes revealed a strong differential impact of particular signal peptidases on the magnitude of the secretion stress response. CONCLUSIONS: The results of the present study highlight the importance of SecDF, SecG and RasP for protein secretion and reveal unexpected differences in the induction of the secretion stress response in different mutant strains. BioMed Central 2020-02-28 /pmc/articles/PMC7048088/ /pubmed/32111210 http://dx.doi.org/10.1186/s12934-020-01315-2 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Neef, Jolanda
Bongiorni, Cristina
Schmidt, Brian
Goosens, Vivianne J.
van Dijl, Jan Maarten
Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title_full Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title_fullStr Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title_full_unstemmed Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title_short Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis
title_sort relative contributions of non-essential sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by bacillus subtilis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7048088/
https://www.ncbi.nlm.nih.gov/pubmed/32111210
http://dx.doi.org/10.1186/s12934-020-01315-2
work_keys_str_mv AT neefjolanda relativecontributionsofnonessentialsecpathwaycomponentsandcellenvelopeassociatedproteasestohighlevelenzymesecretionbybacillussubtilis
AT bongiornicristina relativecontributionsofnonessentialsecpathwaycomponentsandcellenvelopeassociatedproteasestohighlevelenzymesecretionbybacillussubtilis
AT schmidtbrian relativecontributionsofnonessentialsecpathwaycomponentsandcellenvelopeassociatedproteasestohighlevelenzymesecretionbybacillussubtilis
AT goosensviviannej relativecontributionsofnonessentialsecpathwaycomponentsandcellenvelopeassociatedproteasestohighlevelenzymesecretionbybacillussubtilis
AT vandijljanmaarten relativecontributionsofnonessentialsecpathwaycomponentsandcellenvelopeassociatedproteasestohighlevelenzymesecretionbybacillussubtilis