A functionally-distinct carboxylic acid reductase (PcCAR4) unearthed from a repertoire of type IV CARs in the white-rot fungus Pycnoporus cinnabarinus

Jonathan Guyang Ling, Muhamad Hawari Mansor, Munir Abdul Murad, Rozida Mohd. Khalid, Doris Huai-Xia Quay, Margit Winkler, Farah Diba Abu Bakar

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Abstract

Carboxylic acid reductases (CARs) are attracting burgeoning attention as biocatalysts for organic synthesis of aldehydes and their follow-up products from economic carboxylic acid precursors. The CAR enzyme class as a whole, however, is still poorly understood. To date, relatively few CAR sequences have been reported, especially from fungal sources. Here, we sought to increase the diversity of the CAR enzyme class. Six new CAR sequences from the white-rot fungus Pycnoporus cinnabarinus were identified from genome-wide mining. Genome and gene clustering analysis suggests that these PcCARs play different natural roles in Basidiomycete systems, compared to their type II Ascomycete counterparts. The cDNA sequences of all six PcCAR genes were deduced and analysis of their corresponding amino acid sequence showed that they encode for proteins of similar properties and that they possess a conserved modular functional tri-domain arrangement. Phylogenetic analyses showed that all PcCARs cluster with together with the other type IV CARs. One candidate, PcCAR4, was cloned and over-expressed recombinantly in Escherichia coli. Subsequent biotransformation-based screening with a panel of structurally-diverse carboxylic acid substrates suggest that PcCAR4 possessed a more pronounced substrate specificity compared to previously reported CARs. These findings thus present a new functionally-distinct member of the CAR enzyme class.
Originalspracheenglisch
Seitenumfang27
FachzeitschriftJournal of Biotechnology
DOIs
PublikationsstatusAngenommen/In Druck - 2019

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Pycnoporus
Fungi
Carboxylic acids
Genes
Enzymes
Carboxylic Acids
Genome
Synthetic Chemistry Techniques
Basidiomycota
Ascomycota
carboxylic acid reductase
Oxidoreductases
Substrates
Biotransformation
Substrate Specificity
Aldehydes
Biocatalysts
Escherichia coli
Cluster Analysis
Amino Acid Sequence

Fields of Expertise

  • Human- & Biotechnology

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A functionally-distinct carboxylic acid reductase (PcCAR4) unearthed from a repertoire of type IV CARs in the white-rot fungus Pycnoporus cinnabarinus. / Ling, Jonathan Guyang; Mansor, Muhamad Hawari; Murad, Munir Abdul; Khalid, Rozida Mohd. ; Quay, Doris Huai-Xia; Winkler, Margit; Bakar, Farah Diba Abu.

in: Journal of Biotechnology, 2019.

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Ling, Jonathan Guyang ; Mansor, Muhamad Hawari ; Murad, Munir Abdul ; Khalid, Rozida Mohd. ; Quay, Doris Huai-Xia ; Winkler, Margit ; Bakar, Farah Diba Abu. / A functionally-distinct carboxylic acid reductase (PcCAR4) unearthed from a repertoire of type IV CARs in the white-rot fungus Pycnoporus cinnabarinus. in: Journal of Biotechnology. 2019.
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title = "A functionally-distinct carboxylic acid reductase (PcCAR4) unearthed from a repertoire of type IV CARs in the white-rot fungus Pycnoporus cinnabarinus",
abstract = "Carboxylic acid reductases (CARs) are attracting burgeoning attention as biocatalysts for organic synthesis of aldehydes and their follow-up products from economic carboxylic acid precursors. The CAR enzyme class as a whole, however, is still poorly understood. To date, relatively few CAR sequences have been reported, especially from fungal sources. Here, we sought to increase the diversity of the CAR enzyme class. Six new CAR sequences from the white-rot fungus Pycnoporus cinnabarinus were identified from genome-wide mining. Genome and gene clustering analysis suggests that these PcCARs play different natural roles in Basidiomycete systems, compared to their type II Ascomycete counterparts. The cDNA sequences of all six PcCAR genes were deduced and analysis of their corresponding amino acid sequence showed that they encode for proteins of similar properties and that they possess a conserved modular functional tri-domain arrangement. Phylogenetic analyses showed that all PcCARs cluster with together with the other type IV CARs. One candidate, PcCAR4, was cloned and over-expressed recombinantly in Escherichia coli. Subsequent biotransformation-based screening with a panel of structurally-diverse carboxylic acid substrates suggest that PcCAR4 possessed a more pronounced substrate specificity compared to previously reported CARs. These findings thus present a new functionally-distinct member of the CAR enzyme class.",
author = "Ling, {Jonathan Guyang} and Mansor, {Muhamad Hawari} and Murad, {Munir Abdul} and Khalid, {Rozida Mohd.} and Quay, {Doris Huai-Xia} and Margit Winkler and Bakar, {Farah Diba Abu}",
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AU - Mansor, Muhamad Hawari

AU - Murad, Munir Abdul

AU - Khalid, Rozida Mohd.

AU - Quay, Doris Huai-Xia

AU - Winkler, Margit

AU - Bakar, Farah Diba Abu

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