Overproduction of Bacillus amyloliquefaciens extracellular glutamyl-endopeptidase as a result of ectopic multi-copy insertion of an efficiently-expressed mpr gene into the Bacillus subtilis chromosome
2011

Producing More Enzymes with Bacillus subtilis

publication Evidence: moderate

Author Information

Author(s): Yomantas Yurgis AV, Abalakina Elena G, Golubeva Lyubov I, Gorbacheva Lyubov Y, Mashko Sergey V

Primary Institution: Ajinomoto-Genetika Research Institute

Hypothesis

Can a plasmid-less Bacillus subtilis strain produce glutamyl-specific protease more efficiently through multi-copy gene integration?

Conclusion

The study developed a method for integrating multiple copies of a gene into the Bacillus subtilis chromosome, resulting in efficient production of glutamyl-specific protease.

Supporting Evidence

  • The engineered Bacillus subtilis strain produced approximately 0.5 g/L of secreted glutamyl-specific protease.
  • The new method allowed for stable integration of multiple gene copies into the bacterial chromosome.
  • The performance of the new strain was comparable to that of a strain carrying the gene on a multi-copy plasmid.

Takeaway

Scientists figured out how to make a type of bacteria produce more of a special enzyme by putting extra copies of the gene for that enzyme into its DNA.

Methodology

The study involved constructing a plasmid-less Bacillus subtilis strain and integrating multiple copies of the mpr gene into its chromosome using double-cross recombination.

Limitations

The efficiency of gene conversion decreased with an increase in the number of integrated cassettes, which could affect strain stability.

Digital Object Identifier (DOI)

10.1186/1475-2859-10-64

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