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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.).

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 Å resolution after being cooled in a stream of N(2) immediately prior to X-ray diffraction experiments. The crystals belonged to space group C222(1), with unit-cell parameters a = 66.03, b = 183.94, c = 172.28 Å. An initial molecular-replacement solution has been obtained and refinement is in progress.

SUBMITTER: Kuaprasert B

PROVIDER: S-EPMC3212367 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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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.).

Kuaprasert Buabarn B Silprasit Kun K Horata Natharinee N Khunrae Pongsak P Wongpanya Ratree R Boonyalai Nonlawat N Vanavichit Apichart A Choowongkomon Kiattawee K

Acta crystallographica. Section F, Structural biology and crystallization communications 20110929 Pt 10

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 ...[more]

PMID: 22102032

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Project description:The genetic diversity of rice germplasm is the basis for increases in rice yield and quality. The collection, assessment, and protection of the genetic diversity of rice germplasm is important for achieving sustainable agriculture and assuring food security. Many underdeveloped indigenous areas have abundant and valuable rice germplasm resources. However, in-depth assessments of the genetic diversity of rice germplasm from these areas and studies related to protecting these traditional cultures are not available. In this study, from 2005 to 2016, the authors have conducted in-depth evaluation of the genetic diversity of Kam fragrant glutinous rice germplasm resources in southeast Guizhou by using multidisciplinary comprehensive methods such as ethnobotany, cultural anthropology, and modern molecular markers. In total, 376 Kam fragrant glutinous rice samples from 42 villages in the Dong community in southeast Guizhou were collected. Agronomic traits of panicles were complex and exhibited diversity. Some varieties had good disease resistance and adaptation to cold and wet climates. The Dong people named the Kam fragrant glutinous rice varieties by using seven elements, including diverse traits, growth environment, and origin. Traditional folk classification, in addition to morphology and biological analysis using molecular markers, indicates that Kam fragrant glutinous rice includes 91 varieties. Kam fragrant glutinous rice comprises a very high number of varieties, most of which are japonica-type and exhibit a high level of genetic diversity. The traditional folk classification of Kam fragrant glutinous rice by the Dong community is consistent with the biological classification. The traditional naming of Kam fragrant glutinous rice provides an important reference for understanding its genetic diversity. The high level of genetic diversity in Kam fragrant glutinous rice is not only related to the natural environment of the area but also tightly linked with the abundant and diverse Dong ethnic traditional cultures, which has led to protection of Kam fragrant glutinous rice's genetic diversity.

Dataset Information

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.).

Publications

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.).

Similar Datasets

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