| RRC ID |
40782
|
| Author |
Satomura T, Ishikura M, Koyanagi T, Sakuraba H, Ohshima T, Suye S.
|
| Title |
Dye-linked D-amino acid dehydrogenase from the thermophilic bacterium Rhodothermus marinus JCM9785: characteristics and role in trans-4-hydroxy-L-proline catabolism.
|
| Journal |
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.
|
| Volume |
99(10)
|
| Pages |
4265-75
|
| Published |
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
|
| Times Cited |
5
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| Resource |
| General Microbes |
JCM 9785 |
