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, Abdul Munir Abdul Murad, Rozida Mohd. Khalid, Doris Huai Xia Quay, Margit Winkler, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

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 PcCAR enzymes 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 that possess a conserved modular functional tri-domain arrangement. Phylogenetic analyses showed that all PcCAR enzymes cluster 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, preferring to reduce sterically-rigid carboxylic acids such as benzoic acid. These findings thus present a new functionally-distinct member of the CAR enzyme class.

Original languageEnglish
Pages (from-to)55-62
Number of pages8
JournalJournal of Biotechnology
Volume307
DOIs
Publication statusPublished - 10 Jan 2020

Fingerprint

Pycnoporus
Biocatalysis
Cloning
Fungi
Carboxylic acids
Escherichia coli
Organism Cloning
Enzymes
Genes
Substrates
Carboxylic Acids
Genome
Synthetic Chemistry Techniques
Basidiomycota
Ascomycota
Benzoic Acid
carboxylic acid reductase
Oxidoreductases
Biotransformation
Substrate Specificity

Keywords

  • Biocatalysis
  • Carboxylic acid reductase
  • Genome mining
  • Substrate profiling
  • Whole-cell biotransformation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

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; Abdul Murad, Abdul Munir; Mohd. Khalid, Rozida; Quay, Doris Huai Xia; Winkler, Margit; Abu Bakar, Farah Diba.

In: Journal of Biotechnology, Vol. 307, 10.01.2020, p. 55-62.

Research output: Contribution to journalArticle

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