Molecular cloning and functional characterization of two kinds of betaine-aldehyde dehydrogenase in betaine-accumulating mangrove Avicennia marina (Forsk.) Vierh

• Published in: Plant molecular biology 2001-02, Vol.45 (3), p.353-363
• Main Authors: Hibino, T, Meng, Y L, Kawamitsu, Y, Uehara, N, Matsuda, N, Tanaka, Y, Ishikawa, H, Baba, S, Takabe, T, Wada, K, Ishii, T
• Format: Article
• Language: English
• Subjects: Aldehyde Oxidoreductases - genetics; Aldehyde Oxidoreductases - metabolism; Amino Acid Sequence; Avicennia marina; Betaine - metabolism; Betaine-Aldehyde Dehydrogenase; Calcium Chloride - pharmacology; Carbohydrate Metabolism; Cloning; Cloning, Molecular; Dehydrogenase; DNA, Complementary - chemistry; DNA, Complementary - genetics; Dose-Response Relationship, Drug; E coli; Enzyme Stability; Escherichia coli; gamma-Aminobutyric Acid - metabolism; Gene Expression Regulation, Enzymologic - drug effects; Gene Expression Regulation, Plant - drug effects; glycinebetaine; High temperature; Hot Temperature; Isoenzymes - genetics; Molecular Sequence Data; Osmolar Concentration; Oxidation; Oxidation-Reduction - drug effects; Oxygenases - metabolism; Plant Leaves - drug effects; Plant Leaves - enzymology; Plant Leaves - genetics; Plants, Medicinal - enzymology; Plants, Medicinal - genetics; Plants, Medicinal - metabolism; Potassium Chloride - pharmacology; Proline - metabolism; RNA, Messenger - drug effects; RNA, Messenger - genetics; RNA, Messenger - metabolism; Salinity; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Sodium Chloride - pharmacology; Species Specificity; Spinacia oleracea - enzymology; Substrate Specificity; Tissue Distribution
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1023/A:1006497113323

Glycinebetaine is an important osmoprotectant in bacteria, plants, and animals, but only little information is available on the synthesis of glycinebetaine in tree plants. Among four mangrove species, glycinebetaine could be detected only in Avicennia marina. Pinitol was the main osmoprotectant in the other three species. The level of glycinebetaine in A. marina increased under high salinity. Betaine-aldehyde dehydrogenase (BADH) was detected in all four species, but choline monooxygenase could not be detected. A cDNA library was constructed from the leaves of A. marina. Two kinds of BADH cDNAs were isolated, one homologous to the spinach chloroplast BADH, and the other with unique residues SKL at the end of C-terminus. The BADH transcription levels of the former were higher than those of the latter. The levels of the former BADH increased at high salinity whereas those of the latter were independent of salinity. BADHs were expressed in Escherichia coli and purified. Two kinds of A. marina BADHs exhibited similar kinetic and stability properties, but were significantly different from those of spinach BADH. A. marina BADHs efficiently catalyzed the oxidation of betainealdehyde, but not the oxidation of omega-aminoaldehydes and were more stable at high temperature than the spinach BADH.