論文 - 詳細
| RRC ID | 47540 |
|---|---|
| 著者 | Hatada Y, Mizuno M, Li Z, Ohta Y. |
| タイトル | Hyper-production and characterization of the ι-carrageenase useful for ι-carrageenan oligosaccharide production from a deep-sea bacterium, Microbulbifer thermotolerans JAMB-A94T, and insight into the unusual catalytic mechanism. |
| ジャーナル | Mar Biotechnol (NY) |
| Abstract |
A gene of unknown function from the genome of the agar-degrading deep-sea bacterium Microbulbifer thermotolerans JAMB-A94(T) was functionally identified as a ι-carrageenase gene. This gene, designated as cgiA, is located together with two β-agarase genes, agaA and agaO in a cluster. The cgiA gene product is 569 amino acids and shares 29% identity over 185 amino acids with the ι-carrageenase from Zobellia galactanivorans Dsij DSM 12802. Recombinant, cgiA-encoded ι-carrageenase (55 kDa) was hyper-produced in Bacillus subtilis. The recombinant enzyme shows maximal activity at 50°C, the highest reported optimal temperature for a carrageenase. It cleaved β-1,4 linkages in ι-carrageenan to produce a high ratio of ι-carrageenan tetramer, more than 75% of the total product, and did not cleave the β-1,4 linkages in κ- or λ-carrageenan. Therefore, this enzyme may be useful for industrial production of ι-carrageenan oligosaccharides, which have demonstrated antiviral potential against diverse viruses. Furthermore, we performed site-directed mutagenesis on the gene to identify the catalytic amino acid residues. We demonstrated that a conserved Glu351 was essential for catalysis; however, this enzyme lacked a catalytic Asp residue, which is generally critical for the catalytic activity of most glycoside hydrolases. |
| 巻・号 | 13(3) |
| ページ | 411-22 |
| 公開日 | 2011-6-1 |
| DOI | 10.1007/s10126-010-9312-0 |
| PMID | 20686828 |
| MeSH | Alteromonadaceae / chemistry Alteromonadaceae / enzymology* Amino Acid Sequence Bacterial Proteins / genetics* Base Sequence Biotechnology / methods* Carrageenan / biosynthesis* Catalysis Genes, Bacterial / genetics* Glycoside Hydrolases / genetics* Hot Temperature Models, Molecular* Molecular Sequence Data Mutagenesis, Site-Directed Sequence Analysis, DNA Sequence Homology Species Specificity |
| IF | 2.798 |
| 引用数 | 26 |
| WOS 分野 | MARINE & FRESHWATER BIOLOGY 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 | 1 |
| 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 14709 |