Isolation and cloning of an aryl-aldehyde dehydrogenase gene from the white-rot fungus Pycnoporus cinnabarinus strain MUCL 39533

Research output: Contribution to journalArticle

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 Reverse-Transcription-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 super-family. The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.

Original languageEnglish
Pages (from-to)391-397
Number of pages7
JournalMalaysian Journal of Microbiology
Volume11
Issue number4
Publication statusPublished - 2015

Fingerprint

Pycnoporus
Aldehyde Dehydrogenase
Organism Cloning
Fungi
Complementary DNA
Vanillic Acid
Genes
Polymerase Chain Reaction
Trametes
Amino Acids
Proteins
Enzymes
Codon
Computer Simulation
Open Reading Frames
Reverse Transcription
Amino Acid Sequence
Clone Cells
Bacteria

Keywords

  • Aldehyde dehydrogenase super-family
  • Vanillic acid
  • Vanillin

ASJC Scopus subject areas

  • Infectious Diseases
  • Microbiology (medical)

Cite this

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title = "Isolation and cloning of an aryl-aldehyde dehydrogenase gene from the white-rot fungus Pycnoporus cinnabarinus strain MUCL 39533",
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 Reverse-Transcription-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 super-family. The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.",
keywords = "Aldehyde dehydrogenase super-family, Vanillic acid, Vanillin",
author = "Ong, {Khai Lun} and Liew, {Siew Ling} and {Abd. Mutalib}, Sahilah and {Abd. Murad}, {Abdul Munir} and {Abu Bakar}, {Farah Diba}",
year = "2015",
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pages = "391--397",
journal = "Malaysian Journal of Microbiology",
issn = "1823-8262",
publisher = "Malaysian Society for Microbiology",
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TY - JOUR

T1 - Isolation and cloning of an aryl-aldehyde dehydrogenase gene from the white-rot fungus Pycnoporus cinnabarinus strain MUCL 39533

AU - Ong, Khai Lun

AU - Liew, Siew Ling

AU - Abd. Mutalib, Sahilah

AU - Abd. Murad, Abdul Munir

AU - Abu Bakar, Farah Diba

PY - 2015

Y1 - 2015

N2 - 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 Reverse-Transcription-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 super-family. The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.

AB - 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 Reverse-Transcription-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 super-family. The cDNA clone will be used in expression studies to confirm the catalytic function of the enzyme.

KW - Aldehyde dehydrogenase super-family

KW - Vanillic acid

KW - Vanillin

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