Characterization of Cd36_03230p, a putative vanillin dehydrogenase from Candida dubliniensis

Suprama Datta, Uday S. Annapure, David Timson

Research output: Contribution to journalArticle

Abstract

A coding sequence (CD36-03230) from the yeast Candidadubliniensis had been previously annotated as a vanillin dehydrogenase (VDH). The corresponding protein (CD36-03230p) was recombinantly expressed in Escherichia coli and analysed. The protein is most likely a tetramer in solution as judged by crosslinking and gel filtration experiments. CD36-03230p is an active aldehyde dehydrogenase favouring cyclic and aromatic substrates. Positive cooperativity and substrate inhibition wereobserved with some substrates. The redox cofactor NADP+ andsubstrates affected the thermal stability of the protein. Interestingly, the enzyme had no detectable activity with vanillin suggesting that the annotation is incorrect. It has been previously hypothesized that a methionine residue at a key position in the active site of yeast aldehyde dehydrogenases sterically hinders cyclic substrates and restricts specificity to aliphatic aldehydes. Molecular modeling of CD36-03230p demonstrates that it has an isoleucine residue (Ile-156) at this position, further strengtheningthis hypothesis.
Original languageEnglish
Pages (from-to)99774-99780
Number of pages7
JournalRSC Advances
Volume6
Issue number102
DOIs
Publication statusPublished - 14 Oct 2016

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Candida
Aldehyde Dehydrogenase
Substrates
Yeast
CD36 Antigens
Molecular modeling
Isoleucine
NADP
Aldehydes
Methionine
Crosslinking
Escherichia coli
Proteins
Thermodynamic stability
Gels
vanillin dehydrogenase
Enzymes
Experiments

Bibliographical note

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.

Cite this

Datta, Suprama ; Annapure, Uday S. ; Timson, David. / Characterization of Cd36_03230p, a putative vanillin dehydrogenase from Candida dubliniensis. In: RSC Advances. 2016 ; Vol. 6, No. 102. pp. 99774-99780.
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Characterization of Cd36_03230p, a putative vanillin dehydrogenase from Candida dubliniensis. / Datta, Suprama; Annapure, Uday S.; Timson, David.

In: RSC Advances, Vol. 6, No. 102, 14.10.2016, p. 99774-99780.

Research output: Contribution to journalArticle

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T1 - Characterization of Cd36_03230p, a putative vanillin dehydrogenase from Candida dubliniensis

AU - Datta, Suprama

AU - Annapure, Uday S.

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