Genetic improvement of Bacillus licheniformis strains for efficient deproteinization of shrimp shells and production of high-molecular-mass chitin and chitosan

Appl Environ Microbiol. 2010 Dec;76(24):8211-21. doi: 10.1128/AEM.01404-10. Epub 2010 Oct 22.

Abstract

By targeted deletion of the polyglutamate operon (pga) in Bacillus licheniformis F11, a derivative form, F11.1 (Δpga), was obtained that, along with lacking polyglutamate (PGA) formation, displayed enhanced proteolytic activities. The phenotypic properties were maintained in a strain in which the chiBA operon was additionally deleted: F11.4 (ΔchiBA Δpga). These genetically modified strains, carrying the Δpga deletion either alone (F11.1) or together with the ΔchiBA (F11.4) deletion, were used in fermentations (20-liter scale) aiming at the deproteinization of shrimp shells in order to obtain long-chain chitin. After chemical deacetylation, the resulting chitosan samples were analyzed by nuclear magnetic resonance spectroscopy, size exclusion chromatography, and viscometry and compared to a chitosan preparation that was produced in parallel by chemical methods by a commercial chitosan supplier (GSRmbH). Though faint lipid impurities were present in the fermented polysaccharides, the viscosity of the material produced with the double-deletion mutant F11.4 (Δpga ΔchiBA) was higher than that of the chemically produced and commercially available samples (Cognis GmbH). Thus, enhanced proteolytic activities and a lack of chitinase activity render the double mutant F11.4 a powerful tool for the production of long-chain chitosan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus / genetics*
  • Bacillus / metabolism*
  • Chitin / chemistry
  • Chitin / isolation & purification
  • Chitin / metabolism*
  • Chitosan / chemistry
  • Chitosan / isolation & purification
  • Chitosan / metabolism*
  • Chromatography, Gel
  • Genes, Bacterial
  • Lipids / analysis
  • Molecular Weight
  • Organisms, Genetically Modified / genetics
  • Organisms, Genetically Modified / metabolism
  • Penaeidae / microbiology*
  • Polyglutamic Acid / metabolism
  • Sequence Deletion

Substances

  • Lipids
  • Chitin
  • Polyglutamic Acid
  • Chitosan