Isolation of an Escherichia coli K4 kfoC mutant over-producing capsular chondroitin

Microb Cell Fact. 2010 May 17:9:34. doi: 10.1186/1475-2859-9-34.

Abstract

Background: Chondroitin sulphate is a complex polysaccharide having important structural and protective functions in animal tissues. Extracted from animals, this compound is used as a human anti-inflammatory drug. Among bacteria, Escherichia coli K4 produces a capsule containing a non-sulphate chondroitin and its development may provide an efficient and cheap fermentative production of the polysaccharide.

Results: A random N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis was performed on E. coli K4 to isolate mutants showing an increased production of chondroitin. Several mutants were isolated, one of which, here named VZ15, produced about 80% more chondroitin than the wild type E. coli. We found that the mutant has a missense mutation in the codon 313 of kfoC, the gene encoding chondroitin polymerase (K4CP), with a change from arginine to glutamine. A docking analysis to explain the increased productivity of the K4CP enzyme is presented.

Conclusion: The enhanced chondroitin production by the E. coli K4 mutant reported here shows the validity of the strain improvement strategy for more cost-friendly fermentative processes in the production of this pharmaceutically important but so-far expensive polysaccharide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal
  • Bacterial Capsules
  • Chondroitin Sulfates / biosynthesis*
  • Escherichia coli / cytology
  • Escherichia coli / genetics*
  • Escherichia coli Proteins / genetics
  • Gene Expression Regulation, Bacterial
  • Hexosyltransferases / genetics*
  • Mutation, Missense
  • Organisms, Genetically Modified / genetics*

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Escherichia coli Proteins
  • Chondroitin Sulfates
  • Hexosyltransferases
  • chondroitin polymerase, E coli