| RRC ID |
28686
|
| 著者 |
Saito R, Nakayama A.
|
| タイトル |
Differences in malate dehydrogenases from the obligately piezophilic deep-sea bacterium Moritella sp. strain 2D2 and the psychrophilic bacterium Moritella sp. strain 5710.
|
| ジャーナル |
FEMS Microbiol Lett
|
| Abstract |
The gene encoding malate dehydrogenase (MDH) of the obligately piezophilic deep-sea bacterium Moritella sp. strain 2D2 was cloned and sequenced. There were two positions [close to the active site (Ala-180) and in the subunit interaction site (His-229)] with 2D2-specific substitutions. The MDH genes of strain 2D2 and a psychrophilic bacterium Moritella sp. strain 5710 exhibiting the highest sequence similarity were overexpressed in Escherichia coli. The 2D2 MDH was more heat-stable than the 5710 MDH. The apparent Km value at 62.1 MPa for NADH of the 2D2 MDH was higher than that of the 5710 MDH. The 2D2 MDH in which a His-Gln substitution was introduced at position 229 decreased the thermal stability and Km value at 62.1 MPa. The 5710 MDH that was substituted Gln-229 with His increased the thermal stability and Km value at 62.1 MPa. These results indicate that the His residue at position 229 of the 2D2 MDH may play a role in the thermal stability and the MDH function at high pressure.
|
| 巻・号 |
233(1)
|
| ページ |
165-72
|
| 公開日 |
2004-4-1
|
| DOI |
10.1016/j.femsle.2004.02.004
|
| PII |
S0378109704001193
|
| PMID |
15043884
|
| MeSH |
Amino Acid Sequence
Amino Acid Substitution
Cloning, Molecular
Cold Temperature
DNA, Bacterial / chemistry
DNA, Bacterial / isolation & purification
DNA, Ribosomal / chemistry
DNA, Ribosomal / isolation & purification
Enzyme Stability
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Genes, Bacterial
Genes, rRNA
Hydrostatic Pressure
Kinetics
Malate Dehydrogenase / chemistry
Malate Dehydrogenase / genetics
Malate Dehydrogenase / isolation & purification
Malate Dehydrogenase / metabolism*
Molecular Sequence Data
Moritella / enzymology*
Moritella / genetics
Moritella / physiology
Mutagenesis, Site-Directed
Mutation, Missense
Phylogeny
RNA, Ribosomal, 16S / genetics
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Seawater / microbiology*
Sequence Alignment
Sequence Analysis, DNA
|
| IF |
1.987
|
| 引用数 |
10
|
|
WOS 分野
|
MICROBIOLOGY
|
| リソース情報 |
| 遺伝子材料 |
pET41b FactorXa 2D2 MDH (RDB08301)
pET41b FactorXa H229Q 2D2 MDH (RDB08302)
pET41b FactorXa 5710 MDH (RDB08303)
pET41b FactorXa Q229H 5710 MDH (RDB08304) |
