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

Nevin Gül Karagüler; Richard B. Sessions; Anthony R. Clarke

doi:10.1007/s10529-007-9392-8

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

Nevin Gül Karagüler^*, Richard B. Sessions, Anthony R. Clarke

^*Corresponding author for this work

Department of Molecular Biology and Genetics

University Walk

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	1375-1380
Number of pages	6
Journal	Biotechnology Letters
Volume	29
Issue number	9
DOIs	https://doi.org/10.1007/s10529-007-9392-8
Publication status	Published - Sept 2007

Keywords

Candida methylica
Disulphide bonds
Formate dehydrogenase
Modelling

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title = "Effects of disulphide bridges on the activity and stability of the formate dehydrogenase from Candida methylica",

abstract = "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.",

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T1 - Effects of disulphide bridges on the activity and stability of the formate dehydrogenase from Candida methylica

AU - Karagüler, Nevin Gül

AU - Sessions, Richard B.

AU - Clarke, Anthony R.

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AB - 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.

KW - Candida methylica

KW - Disulphide bonds

KW - Formate dehydrogenase

KW - Modelling

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JO - Biotechnology Letters

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

Abstract

Keywords

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