Document Detail


Self-inducible Bacillus subtilis expression system for reliable and inexpensive protein production by high-cell-density fermentation.
MedLine Citation:
PMID:  21803899     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel technically compliant expression system was developed for heterologous protein production in Bacillus subtilis with the aim of increasing product yields at the same time as decreasing production costs. Standard systems involve the positively regulated manP promoter of the mannose operon, which led to relatively high product yields of 5.3% (5.3 g enhanced green fluorescent protein [eGFP] per 100 g cell dry weight [CDW]) but required large quantities of mannose to induce the reactions, thus rendering the system's technical application rather expensive. To improve this situation, mutant B. subtilis strains were used: the ΔmanA (mannose metabolism) strain TQ281 and the ΔmanP (mannose uptake) strain TQ356. The total amount of inducer could be reduced with TQ281, which, however, displayed sensitivity to mannose. An inducer-independent self-induction system was developed with TQ356 to further improve the cost efficiency and product yield of the system, in which glucose prevents induction by carbon catabolite repression. To create optimal self-induction conditions, a glucose-limited process strategy, namely, a fed-batch process, was utilized as follows. The initiation of self-induction at the beginning of the glucose-restricted transition phase between the batch and fed-batch phase of fermentation and its maintenance throughout the glucose-limiting fed-batch phase led to a nearly 3-fold increase of product yield, to 14.6%. The novel B. subtilis self-induction system thus makes a considerable contribution to improving product yield and reducing the costs associated with its technical application.
Authors:
Marian Wenzel; Alexander Müller; Martin Siemann-Herzberg; Josef Altenbuchner
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Publication Detail:
Type:  Journal Article     Date:  2011-07-29
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  77     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2011 Sep 
Date Detail:
Created Date:  2011-09-09     Completed Date:  2011-12-21     Revised Date:  2013-06-28    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  6419-25     Citation Subset:  IM    
Affiliation:
Institut für Industrielle Genetik, Universität Stuttgart, Stuttgart, Germany.
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MeSH Terms
Descriptor/Qualifier:
Bacillus subtilis / genetics*,  metabolism*
Biotechnology / economics,  methods*
Fermentation
Gene Deletion
Gene Expression Regulation, Bacterial*
Genetic Vectors*
Glucose / metabolism
Mannose / metabolism
Recombinant Proteins / biosynthesis*,  genetics
Transcriptional Activation
Chemical
Reg. No./Substance:
0/Recombinant Proteins; 31103-86-3/Mannose; 50-99-7/Glucose
Comments/Corrections

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