1. Isolation of an Escherichia coli K4 kfoC mutant over-producing capsular chondroitin.
- Author
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Zanfardino A, Restaino OF, Notomista E, Cimini D, Schiraldi C, De Rosa M, De Felice M, and Varcamonti M
- Subjects
- Anti-Inflammatory Agents, Non-Steroidal, Bacterial Capsules, Escherichia coli cytology, Escherichia coli Proteins genetics, Gene Expression Regulation, Bacterial, Mutation, Missense, Chondroitin Sulfates biosynthesis, Escherichia coli genetics, Hexosyltransferases genetics, Organisms, Genetically Modified genetics
- 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.
- Published
- 2010
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