Abstract
| Original language | English |
|---|---|
| Article number | 158 |
| Journal | Microbial Cell Factories |
| Volume | 18 |
| Issue number | 1 |
| Number of pages | 16 |
| ISSN | 1475-2859 |
| DOIs | |
| Publication status | Published - 2019 |
Keywords
- Bacillus subtilis
- Recombinant protein production
- Secretion stress
- PrsA
Cite this
}
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 Fischer; Appel, Karen Fuglede; Kilstrup, Mogens; Rasmussen, Michael D.; Nielsen, Allan Kent.
In: Microbial Cell Factories, Vol. 18, No. 1, 158, 2019.Research output: Contribution to journal › Journal article › Research › peer-review
TY - JOUR
T1 - Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
AU - Quesada Ganuza, Ane
AU - Antelo-Varela, Minia
AU - Mouritzen, Jeppe C.
AU - Bartel, Jürgen
AU - Becher, Dörte
AU - Gjermansen, Morten
AU - Hallin, Peter Fischer
AU - Appel, Karen Fuglede
AU - Kilstrup, Mogens
AU - Rasmussen, Michael D.
AU - Nielsen, Allan Kent
PY - 2019
Y1 - 2019
N2 - 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.
AB - 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.
KW - Bacillus subtilis
KW - Recombinant protein production
KW - Secretion stress
KW - PrsA
U2 - 10.1186/s12934-019-1203-0
DO - 10.1186/s12934-019-1203-0
M3 - Journal article
VL - 18
JO - Microbial Cell Factories
JF - Microbial Cell Factories
SN - 1475-2859
IS - 1
M1 - 158
ER -