Effects of disulphide bridges on the activity and stability of the formate dehydrogenase from Candida methylica

Karagueler N. G., Sessions R. B., Clarke A. R.

BIOTECHNOLOGY LETTERS, vol.29, no.9, pp.1375-1380, 2007 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 29 Issue: 9
  • Publication Date: 2007
  • Doi Number: 10.1007/s10529-007-9392-8
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.1375-1380
  • Istanbul Technical University Affiliated: No


Wild-type cmFDH contains no cystines, hence it is a good candidate to test the hypothesis that thermostability can be achieved by introducing new disulphide bridges. Three cysteine double mutants of cmFDH were designed, using a homology model reported previously, to introduce cystine bridges in the C-domain (T169C-T226C) in the N-domain (V88C-V112C) and between the two monomers (M156C-L159C) to form two cystine bridges across the dimer interface. These mutants were constructed and the proteins were over-expressed in E. coli. The mutants V88C-V112C and M156C-L159C lost FDH activity. The mutant T169C-T226C was both less active and less thermostable than wild-type FDH.