Isolation and cloning of an aryl-aldehyde dehydrogenase gene from the white-rot fungus Pycnoporus cinnabarinus strain MUCL 39533
- Author:
Khai Lun Ong
;
Siew Ling Liew
;
Sahilah Abdul Mutalib
;
Abdul Munir Abdul Murad
;
Farah Diba Abu Bakar
- Publication Type:Letter
- Keywords:
Aldehyde dehydrogenase super-family;
vanillin;
vanillic acid
- MeSH:
Vanillic Acid;
Flavoring Agents
- From:Malaysian Journal of Microbiology
2015;11(4):391-397
- CountryMalaysia
- Language:English
-
Abstract:
Aims: The white rot fungus Pycnoporus cinnabarinus MUCL 39533 is able to reduce vanillic acid to vanillin. Reduction of
vanillic acid to vanillin catalysed by the key enzyme aryl-aldehyde dehydrogenase has been reported. Here we report the
isolation and cloning of aryl-aldehyde dehydrogenase from P. cinnabarinus strain MUCL 39533.
Methodology and results: An aryl-aldehyde dehydrogenase gene (PcALDH) was isolated from P. cinnabarinus by
producing a partial cDNA sequence fragment of an aryl-aldehyde dehydrogenase gene through PCR. Degenerate PCR
primers were designed based on codons corresponding to conserved amino acid regions of aryl-aldehyde
dehydrogenases of several fungi and bacteria. The full-length PcALDH cDNA was obtained through ReverseTranscription-Polymerase
Chain Reaction (RT-PCR) and Rapid Amplification cDNA Ends (RACE) PCR. PcALDH cDNA
comprises an open reading frame of 1,506 bp that encodes a protein of 501 amino acids. The PcALDH predicted protein
showed the highest amino acid sequence identity (84%) to ALDH from Trametes versicolor. In silico analysis of PcALDH
indicated that it belongs to the ALDH super-family and Class 3 ALDH.
Conclusion, significance and impact study: PcALDH cDNA was successfully isolated and characterized. Important
motifs identified from the highly conserved PcALDH protein indicated that it belongs to the aldehyde dehydrogenase superfamily.
The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.
- Full text:P020151209444003917002.pdf
