Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase
| Autor: | Karen F. Appel, Ane Quesada-Ganuza, Dörte Becher, Peter Fischer Hallin, Michael Dolberg Rasmussen, Morten Gjermansen, Allan Kent Nielsen, Mogens Kilstrup, Jürgen Bartel, Minia Antelo-Varela, Jeppe C. Mouritzen |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Protein Folding
Lipoproteins lcsh:QR1-502 Heterologous Bioengineering Bacillus subtilis Applied Microbiology and Biotechnology lcsh:Microbiology law.invention 03 medical and health sciences Bacterial Proteins law Secretion stress Extracellular Secretion Amylase 030304 developmental biology 0303 health sciences PrsA biology 030306 microbiology Chemistry Research Membrane Proteins biology.organism_classification Recombinant protein production Recombinant Proteins NIMA-Interacting Peptidylprolyl Isomerase Increased amylase Biochemistry Metabolic Engineering Yield (chemistry) Recombinant DNA biology.protein alpha-Amylases Biotechnology |
| Zdroj: | Microbial Cell Factories Quesada Ganuza, A, Antelo-Varela, M, Mouritzen, J C, Bartel, J, Becher, D, Gjermansen, M, Hallin, P F, Appel, K F, Kilstrup, M, Rasmussen, M D & Nielsen, A K 2019, ' Identification and optimization of PrsA in Bacillus subtilis for improved yield of amylase ', Microbial Cell Factories, vol. 18, no. 1, 158 . https://doi.org/10.1186/s12934-019-1203-0 Microbial Cell Factories, Vol 18, Iss 1, Pp 1-16 (2019) |
| ISSN: | 1475-2859 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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