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| 海洋产纤维素酶草螺菌的筛选及产酶条件优化 |
| Screening of Marine Cellulase Producing Herbaspirillum huttiense and Enzyme Producing Conditions Optimization |
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| DOI：doi:10.3969/j.issn.1005-7021.2020.04.002 |
| 中文关键词: 草螺菌 纤维素酶 筛选 发酵优化 正交实验 |
| 英文关键词: Herbaspirillum huttiense cellulase screen fermentation optimization orthogonal experiment |
| 基金项目:辽宁省自然科学基金项目（20180550858） |
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| 中文摘要: |
| 采用羧甲基纤维素钠（CMC-Na）培养基和刚果红染色法，从大连黑石礁海域潮间带沙泥的混合水样中筛选出1株产纤维素酶的海洋杆菌。进行了形态学与生理生化鉴定和16S rDNA鉴定及构建系统发育进化树，鉴定结果证明，该菌株属于草螺菌（Herbaspirillum huttiense）。单因素实验表明，最优产酶条件为pH 8.5、接种量≥5%、CMC-Na为碳源，蛋白胨为氮源。在单因素实验基础上，选取影响纤维素酶活力较大的3个因素pH、CMC-Na和蛋白胨，利用正交试验优化草螺菌产纤维素酶的最佳条件为pH 8.0、CMC-Na 1.5 g、蛋白胨1.5 g，优化后酶活力提高为10.498 U/mL。筛选的海洋草螺菌，为大连海域首次发现，具有重要的研究意义。 |
| 英文摘要: |
| Adopting sodium carboxymethylcellulose (CMC-Na) medium and Congo red staining method, one cellulase producing marine bacillus strain was isolated from the mixed water sand sample in Heishi (black stone) reef sea area in Dalian. Morphological, physiological biochemical characterization, and 16S rDNA sequences analysis as well as phylogenetic tree construction were carried out, the results of the identification had proved that the strain belonged to Herbaspirillum huttiense. Single factor experiments showed that its optimized fermentation conditions were as follows: pH 8.5, inoculation volume was ≥5% (v/v); carbon source was CMC-Na and nitrogen source was peptone. And three most affecting factors, pH, CMC-Na and peptone were selected for orthogonal experiment, and the optimized fermentation conditions were as follows: pH 8.0, CMC-Na 1.5 g, and peptone 1.5 g, the cellulase activity increased to 10.498 U/mL after the optimization. The screened Herbaspirillum huttiense is the first discovered in Dalian sea area and possessed important significance in future research. |
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