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Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase

BACKGROUND: PrsA is an extracytoplasmic folding catalyst essential in Bacillus subtilis. Overexpression of the native PrsA from B. subtilis has repeatedly lead to increased amylase yields. Nevertheless, little is known about how the overexpression of heterologous PrsAs can affect amylase secretion....

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Autores principales: Quesada-Ganuza, Ane, Antelo-Varela, Minia, Mouritzen, Jeppe C., Bartel, Jürgen, Becher, Dörte, Gjermansen, Morten, Hallin, Peter F., Appel, Karen F., Kilstrup, Mogens, Rasmussen, Michael D., Nielsen, Allan K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6749698/
https://www.ncbi.nlm.nih.gov/pubmed/31530286
http://dx.doi.org/10.1186/s12934-019-1203-0
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author Quesada-Ganuza, Ane
Antelo-Varela, Minia
Mouritzen, Jeppe C.
Bartel, Jürgen
Becher, Dörte
Gjermansen, Morten
Hallin, Peter F.
Appel, Karen F.
Kilstrup, Mogens
Rasmussen, Michael D.
Nielsen, Allan K.
author_facet Quesada-Ganuza, Ane
Antelo-Varela, Minia
Mouritzen, Jeppe C.
Bartel, Jürgen
Becher, Dörte
Gjermansen, Morten
Hallin, Peter F.
Appel, Karen F.
Kilstrup, Mogens
Rasmussen, Michael D.
Nielsen, Allan K.
author_sort Quesada-Ganuza, Ane
collection PubMed
description BACKGROUND: PrsA is an extracytoplasmic folding catalyst essential in Bacillus subtilis. Overexpression of the native PrsA from B. subtilis has repeatedly lead to increased amylase yields. Nevertheless, little is known about how the overexpression of heterologous PrsAs can affect amylase secretion. RESULTS: In this study, the final yield of five extracellular alpha-amylases was increased by heterologous PrsA co-expression up to 2.5 fold. The effect of the overexpression of heterologous PrsAs on alpha-amylase secretion is specific to the co-expressed alpha-amylase. Co-expression of a heterologous PrsA can significantly reduce the secretion stress response. Engineering of the B. licheniformis PrsA lead to a further increase in amylase secretion and reduced secretion stress. CONCLUSIONS: In this work we show how heterologous PrsA overexpression can give a better result on heterologous amylase secretion than the native PrsA, and that PrsA homologs show a variety of specificity towards different alpha-amylases. We also demonstrate that on top of increasing amylase yield, a good PrsA–amylase pairing can lower the secretion stress response of B. subtilis. Finally, we present a new recombinant PrsA variant with increased performance in both supporting amylase secretion and lowering secretion stress.
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spelling pubmed-67496982019-09-23 Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase Quesada-Ganuza, Ane Antelo-Varela, Minia Mouritzen, Jeppe C. Bartel, Jürgen Becher, Dörte Gjermansen, Morten Hallin, Peter F. Appel, Karen F. Kilstrup, Mogens Rasmussen, Michael D. Nielsen, Allan K. Microb Cell Fact Research BACKGROUND: PrsA is an extracytoplasmic folding catalyst essential in Bacillus subtilis. Overexpression of the native PrsA from B. subtilis has repeatedly lead to increased amylase yields. Nevertheless, little is known about how the overexpression of heterologous PrsAs can affect amylase secretion. RESULTS: In this study, the final yield of five extracellular alpha-amylases was increased by heterologous PrsA co-expression up to 2.5 fold. The effect of the overexpression of heterologous PrsAs on alpha-amylase secretion is specific to the co-expressed alpha-amylase. Co-expression of a heterologous PrsA can significantly reduce the secretion stress response. Engineering of the B. licheniformis PrsA lead to a further increase in amylase secretion and reduced secretion stress. CONCLUSIONS: In this work we show how heterologous PrsA overexpression can give a better result on heterologous amylase secretion than the native PrsA, and that PrsA homologs show a variety of specificity towards different alpha-amylases. We also demonstrate that on top of increasing amylase yield, a good PrsA–amylase pairing can lower the secretion stress response of B. subtilis. Finally, we present a new recombinant PrsA variant with increased performance in both supporting amylase secretion and lowering secretion stress. BioMed Central 2019-09-17 /pmc/articles/PMC6749698/ /pubmed/31530286 http://dx.doi.org/10.1186/s12934-019-1203-0 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.
spellingShingle Research
Quesada-Ganuza, Ane
Antelo-Varela, Minia
Mouritzen, Jeppe C.
Bartel, Jürgen
Becher, Dörte
Gjermansen, Morten
Hallin, Peter F.
Appel, Karen F.
Kilstrup, Mogens
Rasmussen, Michael D.
Nielsen, Allan K.
Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title_full Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title_fullStr Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title_full_unstemmed Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title_short Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
title_sort identification and optimization of prsa in bacillus subtilis for improved yield of amylase
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6749698/
https://www.ncbi.nlm.nih.gov/pubmed/31530286
http://dx.doi.org/10.1186/s12934-019-1203-0
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