論文 - 詳細
| RRC ID | 40707 |
|---|---|
| 著者 | Wang C, Luo H, Niu C, Shi P, Huang H, Meng K, Bai Y, Wang K, Hua H, Yao B. |
| タイトル | Biochemical characterization of a thermophilic β-mannanase from Talaromyces leycettanus JCM12802 with high specific activity. |
| ジャーナル | Appl Microbiol Biotechnol |
| Abstract |
Thermophilic β-mannanases are of increasing importance for wide industrial applications. In the current study, gene cloning, functional expression in Pichia pastoris, and characterization of a thermophilic β-mannanase (Man5A) from thermophilic Talaromyces leycettanus JCM12802 are reported. Deduced Man5A exhibits the highest identity with a putative β-mannanase from Talaromyces stipitatus ATCC10500 (70.3 %) and is composed of an N-terminal signal peptide, a fungal-type carbohydrate-binding module (CBM) of family 1, and a catalytic domain of glycosyl hydrolase (GH) family 5 at the C-terminus. Two recombinant proteins with different glycosylation levels, termed Man5A1 (72 kDa) and Man5A2 (60 kDa), were identified after purification. Both enzymes were thermophilic, exhibiting optimal activity at 85-90 °C, and were highly stable at 70 °C. Man5A1 and Man5A2 had a pH optimum of 4.5 and 4.0, respectively, and were highly stable over the broad pH range of 3.0-10.0. Most metal ions and sodium dodecyl sulfate (SDS) had no effect on the enzymatic activities. Man5A1 and Man5A2 exhibited high specific activity (2,160 and 1,800 U/mg, respectively) when using locust bean gum as the substrate. The CBM1 and two key residues D191 and R286 were found to affect Man5A thermostability. Man5A displays a classical four-site-binding mode, hydrolyzing mannooligosaccharides into smaller units, galactomannan into mannose and mannobiose, and glucomanman into mannose, mannobiose, and mannopentaose, respectively. All these properties make Man5A a good candidate for extensive applications in the bioconversion, pulp bleaching, textile, food, and feed industries. |
| 巻・号 | 99(3) |
| ページ | 1217-28 |
| 公開日 | 2015-2-1 |
| DOI | 10.1007/s00253-014-5979-x |
| PMID | 25104029 |
| MeSH | Amino Acid Sequence Cloning, Molecular Enzyme Stability Gene Expression Hydrogen-Ion Concentration Models, Molecular Molecular Sequence Data Molecular Weight Protein Conformation Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism Sequence Analysis, DNA Sequence Homology, Amino Acid Substrate Specificity Talaromyces / enzymology* Talaromyces / genetics Temperature beta-Mannosidase / chemistry beta-Mannosidase / genetics beta-Mannosidase / metabolism* |
| IF | 3.53 |
| 引用数 | 27 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| 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 | 6 |
| 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 | JCM 12802 |