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A new cotton SDR family gene encodes a polypeptide possessing aldehyde reductase and 3-ketoacyl-CoA reductase activities

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Abstract

To understand regulatory mechanisms of cotton fiber development, microarray analysis has been performed for upland cotton (Gossypium hirsutum). Based on this, a cDNA (GhKCR3) encoding a polypeptide belonging to short-chain alcohol dehydrogenase/reductase family was isolated and cloned. It contains an open reading frame of 987 bp encoding a polypeptide of 328 amino acid residues. Following its overexpression in bacterial cells, the purified recombinant protein specifically uses NADPH to reduce a variety of short-chain aldehydes. A fragment between Gly180 and Gly191 was found to be essential for its catalytic activity. Though the GhKCR3 gene shares low sequence similarities to the ortholog of Saccharomyces cerevisiae YBR159w that encodes 3-ketoacyl-CoA reductase (KCR) catalyzing the second step of fatty acid elongation, it was surprisingly able to complement the yeast ybr159wδ mutant. Gas chromatography-mass spectrometry analysis showed that very long-chain fatty acids, especially C26:0, were produced in the ybr159wδ mutant cells expressing GhKCR3. Applying palmitoyl-CoA and malonyl-CoA as substrates, GhKCR3 showed KCR activity in vitro. Quantitative real time-PCR analysis indicated GhKCR3 transcripts accumulated in rapidly elongating fibers, roots, and stems. Our results suggest that GhKCR3 is probably a novel KCR contributing to very long-chain fatty acid biosynthesis in plants.

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Abbreviations

IPTG:

isopropyl-β-D-1-thiogalactopyranoside

KCR:

3-ketoacyl-CoA reductase

MBP:

maltose-binding protein

SDR family:

short-chain alcohol dehydrogenase/reductase family

VLCFA:

very long-chain fatty acid

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Authors and Affiliations

  1. National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, 100871, Beijing, China

    Yu Pang, Wen-Qiang Song, Fang-Yuan Chen & Yong-Mei Qin

Authors
  1. Yu Pang
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  2. Wen-Qiang Song
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  3. Fang-Yuan Chen
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  4. Yong-Mei Qin
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Correspondence to Yong-Mei Qin.

Additional information

Published in Russian in Biokhimiya, 2010, Vol. 75, No. 3, pp. 391–399.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM09-205, January 31, 2010.

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Pang, Y., Song, WQ., Chen, FY. et al. A new cotton SDR family gene encodes a polypeptide possessing aldehyde reductase and 3-ketoacyl-CoA reductase activities. Biochemistry Moscow 75, 320–326 (2010). https://doi.org/10.1134/S0006297910030089

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  • DOI: https://doi.org/10.1134/S0006297910030089

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