論文 - 詳細
| RRC ID | 63327 |
|---|---|
| 著者 | Gu S, Dai X, Xu Z, Niu Q, Jiang J, Liu Y. |
| タイトル | Molecular, structural and biochemical characterization of a novel recombinant chlorophyllase from cyanobacterium Oscillatoria acuminata PCC 6304. |
| ジャーナル | Microb Cell Fact |
| Abstract |
Background
Chlorophyllase catalyzes the hydrolysis of chlorophyll and produces chlorophyllide and phytol. Cyanobacterial chlorophyllases are likely to be more highly heterologously expressed than plant chlorophyllases. A novel recombinant chlorophyllase from the cyanobacterium Oscillatoria acuminata PCC 6304 was successfully expressed in Escherichia coli BL21(DE3).
Results
The putative N-terminal 28-amino-acid signal peptide sequence of O. acuminata chlorophyllase (OaCLH) is essential for its activity, but may confer poor solubility on OaCLH. The C-terminal fusion of a 6 × His tag caused a partial loss of activity in recombinant OaCLH, but an N-terminal 6 × His tag did not destroy its activity. The optimal pH and temperature for recombinant OaCLH activity are 7.0 and 40 °C, respectively. Recombinant OaCLH has hydrolysis activities against chlorophyll a, chlorophyll b, bacteriochlorophyll a, and pheophytin a, but prefers chlorophyll b and chlorophyll a as substrates. The results of site-directed mutagenesis experiments indicated that the catalytic triad of OaCLH consists of Ser159, Asp226, and His258.
Conclusions
The high-level expression and broad substrate specificity of recombinant OaCLH make it suitable for genetically engineering and a promising biocatalyst for industrial production, with applications in vegetable oil refining and laundry detergents. |
| 巻・号 | 20(1) |
| ページ | 14 |
| 公開日 | 2021-1-12 |
| DOI | 10.1186/s12934-020-01507-w |
| PII | 10.1186/s12934-020-01507-w |
| PMID | 33430874 |
| PMC | PMC7802212 |
| MeSH | Amino Acid Sequence Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* Biocatalysis Carboxylic Ester Hydrolases / chemistry Carboxylic Ester Hydrolases / genetics Carboxylic Ester Hydrolases / metabolism* Chlorophyll / metabolism* Chlorophyll A / metabolism* Cloning, Molecular Escherichia coli / genetics Hydrogen-Ion Concentration Hydrolysis Kinetics Models, Molecular Oscillatoria / enzymology* Oscillatoria / genetics Protein Structure, Tertiary Recombinant Proteins / chemistry Recombinant Proteins / metabolism* Sequence Homology, Amino Acid Substrate Specificity Temperature |
| IF | 4.187 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 藻類 | NIES-2108 |