Purification, crystallization and preliminary X-ray analysis of recombinant betaine aldehyde dehydrogenase 2 (OsBADH2), a protein involved in jasmine aroma, from Thai fragrant rice (Oryza sativa L.)
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Author list: Kuaprasert B., Silprasit K., Horata N., Khunrae P., Wongpanya R., Boonyalai N., Vanavichit A., Choowongkomon K.
Publication year: 2011
Volume number: 67
Issue number: 10
Start page: 1221
End page: 1223
Number of pages: 3
ISSN: 1744-3091
eISSN: 1744-3091
Languages: English-Great Britain (EN-GB)
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Abstract
Fragrant rice (Oryza sativa L.) betaine aldehyde dehydrogenase 2 (OsBADH2) is a key enzyme in the synthesis of fragrance aroma compounds. The extremely low activity of OsBADH2 in catalyzing the oxidation of acetaldehyde is believed to be crucial for the accumulation of the volatile compound 2-acetyl-1-pyrroline (2AP) in many scented plants, including fragrant rice. Recombinant fragrant rice OsBADH2 was expressed in Escherichia coli as an N-terminal hexahistidine fusion protein, purified using Ni Sepharose affinity chromatography and crystallized using the microbatch method. Initial crystals were obtained within 24 h using 0.1 M Tris pH 8.5 with 30%(w/v) PEG 4000 and 0.2 M magnesium chloride as the precipitating agent at 291 K. Crystal quality was improved when the enzyme was cocrystallized with NAD +. Improved crystals were grown in 0.1 M HEPES pH 7.4, 24%(w/v) PEG 4000 and 0.2 M ammonium chloride and diffracted to beyond 2.95 ล resolution after being cooled in a stream of N2 immediately prior to X-ray diffraction experiments. The crystals belonged to space group C2221, with unit-cell parameters a = 66.03, b = 183.94, c = 172.28 ล. An initial molecular-replacement solution has been obtained and refinement is in progress. ฉ 2011 International Union of Crystallography All rights reserved.
Keywords
betaine aldehyde dehydrogenases, fragrant rice, Oryza sativa L., OsBADH2
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