Intramembrane protease RasP boosts protein production in Bacillus

Autor: Jan Maarten van Dijl, Cristina Bongiorni, Jolanda Neef, Vivianne J. Goosens, Brian Schmidt
Přispěvatelé: Microbes in Health and Disease (MHD), Translational Immunology Groningen (TRIGR)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
ALPHA-AMYLASE
medicine.medical_treatment
lcsh:QR1-502
Bacillus
Bacillus subtilis
SUBTILIS
Applied Microbiology and Biotechnology
lcsh:Microbiology
NUCLEOTIDE-SEQUENCE
Bacillus licheniformis
Secretory Pathway
biology
SIGNAL PEPTIDASE-I
RasP
Protein Transport
Biochemistry
SECRETORY PROTEINS
ESCHERICHIA-COLI
Biotechnology
EXPRESSION
Proteases
Bacillus amyloliquefaciens
GRAM-POSITIVE BACTERIA
Intramembrane protease
030106 microbiology
CELL-DIVISION
Bioengineering
Protein Sorting Signals
03 medical and health sciences
Bacterial Proteins
medicine
Secretion
Site-2 protease
Serine protease
Protease
Research
Bacillus clausii
Cell Membrane
Amylase
Gene Expression Regulation
Bacterial
biology.organism_classification
GENE
030104 developmental biology
biology.protein
alpha-Amylases
Peptide Hydrolases
Zdroj: Microbial Cell Factories, 16(57). BioMed Central Ltd.
Microbial Cell Factories, Vol 16, Iss 1, Pp 1-9 (2017)
Microbial Cell Factories
ISSN: 1475-2859
Popis: Background: The microbial cell factory Bacillus subtilis is a popular industrial platform for high- level production of secreted technical enzymes. Nonetheless, the effective secretion of particular heterologous enzymes remains challenging. Over the past decades various studies have tackled this problem, and major improvements were achieved by optimizing signal peptides or removing proteases involved in product degradation. On the other hand, serious bottlenecks in the protein export process per se remained enigmatic, especially for protein secretion at commercially significant levels by cells grown to high density. The aim of our present study was to assess the relevance of the intramembrane protease RasP for high- level protein production in B. subtilis.Results: Deletion of the rasP gene resulted in reduced precursor processing and extracellular levels of the overproduced a- amylases AmyE from B. subtilis and AmyL from Bacillus licheniformis. Further, secretion of the overproduced serine protease BPN' from Bacillus amyloliquefaciens was severely impaired in the absence of RasP. Importantly, overexpression of rasP resulted in threefold increased production of a serine protease from Bacillus clausii, and 2.5- to 10-fold increased production of an AmyAc alpha-amylase from Paenibacillus curdlanolyticus, depending on the culture conditions. Of note, growth defects due to overproduction of the two latter enzymes were suppressed by rasP- overexpression.Conclusion: Here we show that an intramembrane protease, RasP, sets a limit to high- level production of two secreted heterologous enzymes that are difficult to produce in the B. subtilis cell factory. This finding was unexpected and suggests that proteolytic membrane sanitation is key to effective enzyme production in Bacillus.
Databáze: OpenAIRE
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