1. A novel strategy to improve protein secretion via overexpression of the SppA signal peptide peptidase in Bacillus licheniformis.
- Author
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Cai D, Wang H, He P, Zhu C, Wang Q, Wei X, Nomura CT, and Chen S
- Subjects
- Aspartic Acid Endopeptidases metabolism, Bacillus licheniformis enzymology, Bacillus licheniformis metabolism, Biomass, Protein Sorting Signals genetics, Protein Transport, Serine Endopeptidases genetics, Serine Endopeptidases metabolism, Subtilisins genetics, Transcription, Genetic, alpha-Amylases genetics, Aspartic Acid Endopeptidases genetics, Bacillus licheniformis genetics, Gene Expression, Subtilisins metabolism, alpha-Amylases metabolism
- Abstract
Background: Signal peptide peptidases play an important role in the removal of remnant signal peptides in the cell membrane, a critical step for extracellular protein production. Although these proteins are likely a central component for extracellular protein production, there has been a lack of research on whether protein secretion could be enhanced via overexpression of signal peptide peptidases., Results: In this study, both nattokinase and α-amylase were employed as prototypical secreted target proteins to evaluate the function of putative signal peptide peptidases (SppA and TepA) in Bacillus licheniformis. We observed dramatic decreases in the concentrations of both target proteins (45 and 49%, respectively) in a sppA deficient strain, while the extracellular protein yields of nattokinase and α-amylase were increased by 30 and 67% respectively in a strain overexpressing SppA. In addition, biomass, specific enzyme activities and the relative gene transcriptional levels were also enhanced due to the overexpression of sppA, while altering the expression levels of tepA had no effect on the concentrations of the secreted target proteins., Conclusions: Our results confirm that SppA, but not TepA, plays an important functional role for protein secretion in B. licheniformis. Our results indicate that the sppA overexpression strain, B. licheniformis BL10GS, could be used as a promising host strain for the industrial production of heterologous secreted proteins.
- Published
- 2017
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