Cloning and Functional Analysis of a Novel Aldo-Keto Reductase from Aloe arborescens

A novel aldo-keto reductase (AKR) was cloned and sequenced from roots of Aloe arborescens by a combination of RT-PCR using degenerate primers based on the conserved sequences of plant polyketide reductases (PKRs) and cDNA library screening by oligonucleotide hybridization. A. arborescens AKR share s...

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Bibliographic Details
Published inBiological & Pharmaceutical Bulletin Vol. 30; no. 12; pp. 2262 - 2267
Main Authors Morita, Hiroyuki, Mizuuchi, Yuusuke, Abe, Tsuyoshi, Kohno, Toshiyuki, Noguchi, Hiroshi, Abe, Ikuro
Format Journal Article
LanguageEnglish
Published Japan The Pharmaceutical Society of Japan 01.12.2007
Pharmaceutical Society of Japan
Japan Science and Technology Agency
Subjects
Alcohol Oxidoreductases - biosynthesis
Alcohol Oxidoreductases - genetics
Alcohol Oxidoreductases - metabolism
Aldehyde Reductase
aldo-keto reductase
Aldo-Keto Reductases
Aloe
Aloe - enzymology
Amino Acid Sequence
Cloning, Molecular
DNA Primers
DNA, Complementary - biosynthesis
DNA, Complementary - genetics
DNA, Plant - biosynthesis
DNA, Plant - genetics
Escherichia coli
Gene Library
Hordeum vulgare
Kinetics
Models, Molecular
Molecular Sequence Data
NADP - metabolism
Oligonucleotides
oxidoreductase
polyketide reductase
Reverse Transcriptase Polymerase Chain Reaction
Spectrophotometry, Ultraviolet
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Summary:A novel aldo-keto reductase (AKR) was cloned and sequenced from roots of Aloe arborescens by a combination of RT-PCR using degenerate primers based on the conserved sequences of plant polyketide reductases (PKRs) and cDNA library screening by oligonucleotide hybridization. A. arborescens AKR share similarities with known plant AKRs (40—66% amino acid sequence identity), maintaining most of the active-site residues conserved in the AKR superfamily enzymes. Interestingly, despite the sequence similarity with PKRs, recombinant enzyme expressed in Escherichia coli did not exhibit any detectable PKR activities. Instead, A. arborescens AKR catalyzed NADPH-dependent reduction of various carbonyl compounds including benzaldehyde and DL-glyceraldehyde. Finally, a homology model on the basis of the crystal structure of Hordeum vulgare AKR predicted the active-site architecture of the enzyme.
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ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.30.2262