RRC ID 40782
著者 Satomura T, Ishikura M, Koyanagi T, Sakuraba H, Ohshima T, Suye S.
タイトル Dye-linked D-amino acid dehydrogenase from the thermophilic bacterium Rhodothermus marinus JCM9785: characteristics and role in trans-4-hydroxy-L-proline catabolism.
ジャーナル Appl Microbiol Biotechnol
Abstract A gene from the thermophilic Gram-negative bacterium Rhodothermus marinus JCM9785, encoding a dye-linked D-amino acid dehydrogenase homologue, was overexpressed in Escherichia coli, and its product was purified and characterized. The expressed enzyme was a highly thermostable dye-linked D-amino acid dehydrogenase that retained more than 80% of its activity after incubation for 10 min at up to 70 °C. When enzyme-catalyzed dehydrogenation of several D-amino acids was carried out using 2,6-dichloroindophenol as the electron acceptor, D-phenylalanine was the most preferable substrate among the D-amino acids tested. Immediately upstream of the dye-linked D-amino acid dehydrogenase gene (dadh) was a gene encoding a 4-hydroxyproline 2-epimerase homologue (hypE). That gene was successfully expressed in E. coli, and the gene product exhibited strong 4-hydroxyproline 2-epimerase activity. Reverse transcription PCR and quantitative real-time PCR showed that the six genes containing the dadh and hypE genes were arranged in an operon and were required for catabolism of trans-4-hydroxy-L-proline in R. marinus. This is the first description of a dye-linked D-amino acid dehydrogenase (Dye-DADH) with broad substrate specificity involved in trans-4-hydroxy-L-proline catabolism.
巻・号 99(10)
ページ 4265-75
公開日 2015-5-1
DOI 10.1007/s00253-014-6263-9
PMID 25472442
MeSH Amino Acid Oxidoreductases / chemistry Amino Acid Oxidoreductases / genetics Amino Acid Oxidoreductases / metabolism* Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* Enzyme Stability Escherichia coli / genetics Escherichia coli / metabolism Hydrogen-Ion Concentration Isomerism Kinetics Proline / chemistry Proline / metabolism* Rhodothermus / chemistry Rhodothermus / enzymology* Rhodothermus / genetics Substrate Specificity
IF 3.53
引用数 5
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
一般微生物 JCM 9785