Characterization of a new acidic NAD+-dependent formate dehydrogenase from thermophilic fungus Chaetomium thermophilum

Gülşah Özgün; Nevin Gül Karagüler; Ossi Turunen; Nicholas J. Turner; Bar Iş Binay

doi:10.1016/j.molcatb.2015.09.014

Characterization of a new acidic NAD⁺-dependent formate dehydrogenase from thermophilic fungus Chaetomium thermophilum

Gülşah Özgün, Nevin Gül Karagüler, Ossi Turunen, Nicholas J. Turner, Bar Iş Binay^*

^*Corresponding author for this work

Department of Molecular Biology and Genetics

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

11 Citations (Scopus)

Abstract

NAD⁺-dependent formate dehydrogenase (FDH) enzyme catalyses the NAD(P)⁺-dependent interconversion of formate anion to carbon dioxide coupled with the conversion of NAD(P)⁺ to NAD(P)H. NADH is widely used as cofactor by enzymes utilized in the biocatalytic synthesis of chiral pharmaceutical intermediates and chemical compounds. For this reason, FDH is used in the enzymatic synthesis of optically active prochiral carbonyl compounds in order to regenerate the NADH. For wider applicability in industrial usage of FDH, there is a demand for higher stability of the enzyme. We report here the overexpression in Escherichia coli and subsequent characterization of FDH from thermophilic fungus Chaetomium thermophilum (CtFDH). CtFDH was found to be more acidic than the typical FDHs from various sources, having its optimum at pH 5, and it also showed moderate thermostability at pH 5. The modelling of the enzyme and formate in the active site indicated that the enzyme functions with metal-independent hydride shift in the same way as the other NAD⁺-dependent FDHs of this enzyme group. Dimer interface properties were also analyzed. CtFDH shows potential being a template for engineering higher thermostability for applications in acidic conditions.

Original language	English
Pages (from-to)	212-217
Number of pages	6
Journal	Journal of Molecular Catalysis - B Enzymatic
Volume	122
DOIs	https://doi.org/10.1016/j.molcatb.2015.09.014
Publication status	Published - 1 Dec 2015

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V. All rights reserved.

Funding

This work was supported by a grant from the TUBİTAK (grant number: 214Z064 ), COST Action CM1303 ⿿Systems Biocatalysis⿿ and special thanks to Prof. Dr. Ed Hurt from Heidelberg University for the strain of C. thermophilum.

Funders	Funder number
TUBİTAK	214Z064
European Cooperation in Science and Technology	CM1303
Universität Heidelberg

Keywords

Biochemical and kinetic characterization
Chaetomium thermophilum
Computational modeling
NAD-dependent acidic formate dehydrogenase
Thermostability

Access to Document

10.1016/j.molcatb.2015.09.014

Cite this

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title = "Characterization of a new acidic NAD+-dependent formate dehydrogenase from thermophilic fungus Chaetomium thermophilum",

abstract = "NAD+-dependent formate dehydrogenase (FDH) enzyme catalyses the NAD(P)+-dependent interconversion of formate anion to carbon dioxide coupled with the conversion of NAD(P)+ to NAD(P)H. NADH is widely used as cofactor by enzymes utilized in the biocatalytic synthesis of chiral pharmaceutical intermediates and chemical compounds. For this reason, FDH is used in the enzymatic synthesis of optically active prochiral carbonyl compounds in order to regenerate the NADH. For wider applicability in industrial usage of FDH, there is a demand for higher stability of the enzyme. We report here the overexpression in Escherichia coli and subsequent characterization of FDH from thermophilic fungus Chaetomium thermophilum (CtFDH). CtFDH was found to be more acidic than the typical FDHs from various sources, having its optimum at pH 5, and it also showed moderate thermostability at pH 5. The modelling of the enzyme and formate in the active site indicated that the enzyme functions with metal-independent hydride shift in the same way as the other NAD+-dependent FDHs of this enzyme group. Dimer interface properties were also analyzed. CtFDH shows potential being a template for engineering higher thermostability for applications in acidic conditions.",

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AU - Karagüler, Nevin Gül

AU - Turunen, Ossi

AU - Turner, Nicholas J.

AU - Binay, Bar Iş

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AB - NAD+-dependent formate dehydrogenase (FDH) enzyme catalyses the NAD(P)+-dependent interconversion of formate anion to carbon dioxide coupled with the conversion of NAD(P)+ to NAD(P)H. NADH is widely used as cofactor by enzymes utilized in the biocatalytic synthesis of chiral pharmaceutical intermediates and chemical compounds. For this reason, FDH is used in the enzymatic synthesis of optically active prochiral carbonyl compounds in order to regenerate the NADH. For wider applicability in industrial usage of FDH, there is a demand for higher stability of the enzyme. We report here the overexpression in Escherichia coli and subsequent characterization of FDH from thermophilic fungus Chaetomium thermophilum (CtFDH). CtFDH was found to be more acidic than the typical FDHs from various sources, having its optimum at pH 5, and it also showed moderate thermostability at pH 5. The modelling of the enzyme and formate in the active site indicated that the enzyme functions with metal-independent hydride shift in the same way as the other NAD+-dependent FDHs of this enzyme group. Dimer interface properties were also analyzed. CtFDH shows potential being a template for engineering higher thermostability for applications in acidic conditions.

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