論文 - 詳細
| RRC ID | 66681 |
|---|---|
| 著者 | Wang Z, Li X, Tian J, Chu Y, Tian Y. |
| タイトル | Cloning, heterologous expression and characterization of a novel streptomyces trypsin in Bacillus subtilis SCK6. |
| ジャーナル | Int J Biol Macromol |
| Abstract |
A novel streptomyces trypsin GM2938 was selected as the object of study. The active GM2938 contains 223 amino acid residues. Constructing recombinant plasmid and transforming Bacillus subtilis SCK6, the heterogenous expression of GM2938 was achieved. Through optimization of fermentation conditions, the expression level of GM2938 reached 1622.2 U/mL (esterase activity) and 33.8 U/mL (amidase activity). The recombinant trypsin was purified and measured: the specific activity of esterase was 5.6 × 103 U/mg, and the specific activity of amidase was 1.1 × 103 U/mg. Furthermore, the enzymatic properties of GM2938 were explore: the optimal reaction temperature and pH were 50 °C and 9.0, respectively; the recombinant enzyme show high stability at 25 °C and range of pH 5.0-9.0; Ca2+, K+, Mg2+, EDTA, DTT, DMSO, methanol, glycerin and ethanediol could promote the esterase and amidase activities at the investigated concentrations, while Fe2+, SDS, tritonx-100, acetone, chloroform and n-hexane inhibited the trypsin activities. Kinetic parameters of GM2938 were calculated: the Km of BAEE was 3.15 × 10-5 mol·L-1, Vmax value was 2.87 × 10-4 mol·L-1·min-1; the Km of BAPAN was 2.20 × 10-4 mol·L-1, the Vmax was 2.40 × 10-4 mol·L-1·min-1. These properties give trypsin GM2938 a potential application prospect. |
| 巻・号 | 147 |
| ページ | 890-897 |
| 公開日 | 2020-3-15 |
| DOI | 10.1016/j.ijbiomac.2019.09.248 |
| PII | S0141-8130(19)35613-2 |
| PMID | 31739056 |
| MeSH | Bacillus subtilis* / enzymology Bacillus subtilis* / genetics Bacterial Proteins* / biosynthesis Bacterial Proteins* / chemistry Bacterial Proteins* / genetics Bacterial Proteins* / isolation & purification Cloning, Molecular* Recombinant Proteins / biosynthesis Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Streptomyces* / enzymology Streptomyces* / genetics Trypsin* / biosynthesis Trypsin* / chemistry Trypsin* / genetics Trypsin* / isolation & purification |
| 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 | 3 |
| 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 | |
| 一般微生物 | JCM32175 |