Journal Article

Synthesis of a Novel Class of Polyhydroxyalkanoates in Arabidopsis Peroxisomes, and Their Use in Monitoring Short-Chain-Length Intermediates of β-Oxidation

Yuko Arai, Hideo Nakashita, Yoshikatu Suzuki, Yumiko Kobayashi, Toshiyuki Shimizu, Michiko Yasuda, Yoshiharu Doi and Isamu Yamaguchi

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 5, pages 555-562
Published in print May 2002 | ISSN: 0032-0781
Published online May 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf068
Synthesis of a Novel Class of Polyhydroxyalkanoates in Arabidopsis Peroxisomes, and Their Use in Monitoring Short-Chain-Length Intermediates of β-Oxidation

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The poly[(R)-3-hydroxyalkanoate] (PHA) synthase gene (phaCAc) of Aeromonas caviae FA440 was modified by adding a peroxisome targeting signal encoding the last 10 amino acids at the carboxyl-terminal of spinach glycolate oxidase. The modified gene was introduced into Arabidopsis thaliana plants by Agrobacterium-mediated transformation. The transgenic Arabidopsis plant expressed the introduced gene and its protein, and it accumulated PHA in its tissues. Gas chromatography-mass spectrometry analysis demonstrated the accumulation of a novel type of PHA, poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate). This strongly suggests that short-chain-length (R)-3-hydroxyacyl-CoAs were generated from intermediates of peroxisomal β-oxidation. It was revealed by using this transgenic plant that Tween-20 can activate peroxisomal β-oxidation of short-chain-length fatty acids.

Keywords: Key words: Arabidopsis thaliana — Peroxisome — β-oxidation — Short-chain-length poly[(R)-3-hydroxyalkanoates] — Transgenic plant.; Abbreviations: PHA, poly[(R)-3-hydroxyalkanoate]; PHB, poly[(R)-3-hydroxybutyrate]; SCL, short-chain-length; MCL, medium-chain-length; LCL, long-chain-length; SC, short-chain; MC, medium-chain; LC, long-chain; 3HA, 3-hydroxyalkanoate; 3HB, 3-hydroxybutyrate; 3HV, 3-hydroxyvalerate; 3HH, 3-hydroxyhexanoate; ACOX, acyl-CoA oxidase; GC-MS, gas chromatography-mass spectrometry; HRP, horseradish peroxidase; Nos-T, Agrobacterium nopaline synthase terminator; PCR, polymerase chain reaction; Tween-20, polyethylene sorbitan monolaurate.

Journal Article.  4388 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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