| RRC ID |
41260
|
| 著者 |
Chen H, Zhang T, Sun T, Ni Z, Le Y, Tian R, Chen Z, Zhang C.
|
| タイトル |
Clostridium thermocellum Nitrilase Expression and Surface Display on Bacillus subtilis Spores.
|
| ジャーナル |
J Mol Microbiol Biotechnol
|
| Abstract |
Nitrilases are an important class of industrial enzymes. They require mild reaction conditions and are highly efficient and environmentally friendly, so they are used to catalyze the synthesis of carboxylic acid from nitrile, a process considered superior to conventional chemical syntheses. Nitrilases should be immobilized to overcome difficulties in recovery after the reaction and to stabilize the free enzyme. The nitrilase from Clostridium thermocellum was expressed, identified and displayed on the surface of Bacillus subtilis spores by using the spore coat protein G of B. subtilis as an anchoring motif. In a free state, the recombinant nitrilase catalyzed the conversion of 3-cyanopyridine to niacin and displayed maximum catalytic activity (8.22 units/mg protein) at 40 °C and pH 7.4. SDS-PAGE and Western blot were used to confirm nitrilase display. Compared with the free enzyme, the spore-immobilized nitrilase showed a higher tolerance for adverse environmental conditions. After the reaction, recombinant spores were recovered via centrifugation and reused 3 times to catalyze the conversion of 3-cyanopyridine with 75.3% nitrilase activity. This study demonstrates an effective means of nitrilase immobilization via spore surface display, which can be applied in biological processes or conversion.
|
| 巻・号 |
25(6)
|
| ページ |
381-7
|
| 公開日 |
2015-1-1
|
| DOI |
10.1159/000441642
|
| PII |
000441642
|
| PMID |
26629931
|
| MeSH |
Aminohydrolases / biosynthesis*
Aminohydrolases / genetics
Aminohydrolases / metabolism
Bacillus subtilis / chemistry
Bacillus subtilis / enzymology*
Bacillus subtilis / genetics
Bacterial Proteins / biosynthesis*
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Cell Wall / metabolism
Clostridium thermocellum / enzymology*
Clostridium thermocellum / genetics
Electrophoresis, Polyacrylamide Gel
Enzyme Activation
Enzyme Stability
Enzymes, Immobilized / biosynthesis*
Enzymes, Immobilized / chemistry
Enzymes, Immobilized / genetics
Escherichia coli / enzymology
Escherichia coli / genetics
Genes, Bacterial
Hydrogen-Ion Concentration
Industrial Microbiology
Niacin / metabolism
Pyridines / metabolism
Recombinant Fusion Proteins / biosynthesis
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Spores, Bacterial / chemistry
Spores, Bacterial / enzymology
Spores, Bacterial / metabolism
|
| IF |
0.803
|
| 引用数 |
8
|
|
WOS 分野
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 9323 |
