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


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.

SUBMITTER: Quesada-Ganuza A 

PROVIDER: S-EPMC6749698 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Publications

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

Quesada-Ganuza Ane A   Antelo-Varela Minia M   Mouritzen Jeppe C JC   Bartel Jürgen J   Becher Dörte D   Gjermansen Morten M   Hallin Peter F PF   Appel Karen F KF   Kilstrup Mogens M   Rasmussen Michael D MD   Nielsen Allan K AK  

Microbial cell factories 20190917 1


<h4>Background</h4>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.<h4>Results</h4>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 heterolog  ...[more]

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