| 摘要: |
| 从青岛周边沿海海藻中分离到一株降解卡拉胶的菌株c.2.1，通过形态学观察和16S rDNA序列分析对其进行鉴定，鉴定为食纤维菌属（Cellulophaga sp.c.2.1）。将其卡拉胶硫酸酯酶（sulA）基因克隆到外源表达载体pET-28a上，并转化到大肠杆菌BL21（DE3）中进行表达。卡拉胶硫酸酯酶基因序列全长1455bp，编码484个氨基酸，该酶的分子量为55.1kDa。红外光谱法分析表明，经该酶酶解后κ-卡拉胶的硫酸基含量有显著降低。 |
| 关键词: 食纤维菌 硫酸酯酶 克隆表达 |
| DOI： |
| 分类号: |
| 基金项目:国家自然基金面上项目(31670002) |
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| Screening the carrageenan hydrolyzing bacterium, exogenous expression of carrageenan sulfate esterase and enzymatic products analysis |
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QIN Fang, GENG Li-Ha, ZHANG Quan-Bin, ZHANG De-Chao
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Institute of Oceanology,Chinese Academy of Sciences
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| Abstract: |
| The carrageenan hydrolyzing marine strain c.2.1 was isolated from coastal algae samples around Qingdao city. The isolated strain was identified by morphology observation and 16S rDNA sequencing. It was identified as Cellulophaga sp. c.2.1. The carrageenan sulfatase (sulA) gene was cloned into the exogenous vector pET-28a and transformed into E. coli BL21 (DE3) for expression. The gene comprised an open reading frame of 1455 bp encoding 484 amino acids. The deduced protein had a calculated molecular weight of 55.1 kDa. Infrared analysis of the enzymatic products showed that the sulfate content of κ-carrageenan was significantly reduced. |
| Key words: Cellulophaga sulfatase cloning and expression |