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

Ane Quesada Ganuza, Minia Antelo-Varela, Jeppe C. Mouritzen, Jürgen Bartel, Dörte Becher, Morten Gjermansen, Peter Fischer Hallin, Karen Fuglede Appel, Mogens Kilstrup, Michael D. Rasmussen, Allan Kent Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

151 Downloads (Pure)

Abstract

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.
Original languageEnglish
Article number158
JournalMicrobial Cell Factories
Volume18
Issue number1
Number of pages16
ISSN1475-2859
DOIs
Publication statusPublished - 2019

Keywords

  • Bacillus subtilis
  • Recombinant protein production
  • Secretion stress
  • PrsA

Fingerprint Dive into the research topics of 'Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase'. Together they form a unique fingerprint.

Cite this