| RRC ID |
47594
|
| 著者 |
Liu J, Asano Y, Ikoma K, Yamashita S, Hirose Y, Shimoyama T, Takahashi S, Nakayama T, Nishino T.
|
| タイトル |
Purification, characterization, and primary structure of a novel N-acyl-D-amino acid amidohydrolase from Microbacterium natoriense TNJL143-2.
|
| ジャーナル |
J Biosci Bioeng
|
| Abstract |
A novel N-acyl-D-amino acid amidohydrolase (DAA) was purified from the cells of a novel species of the genus Microbacterium. The purified enzyme, termed AcyM, was a monomeric protein with an apparent molecular weight of 56,000. It acted on N-acylated hydrophobic D-amino acids with the highest preference for N-acetyl-D-phenylalanine (NADF). Optimum temperature and pH for the hydrolysis of NADF were 45°C and pH 8.5, respectively. The k(cat) and K(m) values for NADF were 41 s⁻¹ and 2.5 mM at 37°C and pH 8.0, although the enzyme activity was inhibited by high concentrations of NADF. Although many known DAAs are inhibited by 1 mM EDTA, AcyM displayed a 65% level of its full activity even in the presence of 20 mM EDTA. Based on partial amino acid sequences of the purified enzyme, the full-length AcyM gene was cloned and sequenced. It encoded a protein of 495 amino acids with a relatively low sequence similarity to a DAA from Alcaligenes faecalis DA1 (termed AFD), a binuclear zinc enzyme of the α/β-barrel amidohydrolase superfamily. The unique cysteine residue that serves as a ligand to the active-site zinc ions in AFD and other DAAs was not conserved in AcyM and was replaced by alanine. AcyM was the most closely related to a DAA of Gluconobacter oxydans (termed Gox1177) and phylogenetically distant from AFD and all other DAAs that have been biochemically characterized thus far. AcyM, along with Gox1177, appears to represent a new phylogenetic subcluster of DAAs.
|
| 巻・号 |
114(4)
|
| ページ |
391-7
|
| 公開日 |
2012-10-1
|
| DOI |
10.1016/j.jbiosc.2012.05.015
|
| PII |
S1389-1723(12)00211-3
|
| PMID |
22721690
|
| MeSH |
Actinomycetales / enzymology*
Amidohydrolases / chemistry*
Amidohydrolases / genetics
Amidohydrolases / isolation & purification*
Amidohydrolases / metabolism
Amino Acid Sequence
Amino Acids / biosynthesis*
Amino Acids / chemistry
Amino Acids / genetics
Amino Acids / isolation & purification
Cloning, Molecular
Industrial Microbiology*
Molecular Sequence Data
Molecular Weight
Phylogeny
Sequence Alignment
Temperature
|
| IF |
2.366
|
| 引用数 |
5
|
|
WOS 分野
|
FOOD SCIENCE & TECHNOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 12611 |
