| 摘要: |
| 采用响应面法对1株高产琼胶酶的印度尼西亚热泉菌Bacillus sp. BI-3的发酵条件进行优化。首先以温度、pH、不同碳源、不同氮源、不同金属离子及接种量作为唯一变量进行单因素实验, 筛选出对酶活有显著影响的单因素取值范围; 参考单因素实验结果, 采用Plackett-Burman实验设计筛选出影响酶活主要因素, 再利用Box-Behnken设计及响应面分析法进行回归分析以确定最佳发酵条件。结果表明, D-甘露糖、氯化锶和氯化钙与酶活存在着显著的相关性, 通过求解回归方程得到Bacillus sp. BI-3的发酵条件系统优化的结果为: 酵母粉0.3%、D-甘露糖0.66%、蛋白胨0.6%、氯化钙7.21mmol/L、氯化锶6.00mmol/L、氯化钠添加量6‰、培养温度为55℃、接种量1%、初始pH 6.0, 优化后发酵上清液的酶活达到6.0U/mL, 比优化前提高了2.4倍。 |
| 关键词: 琼胶酶 嗜热菌 响应面法 优化 |
| DOI：10.11693/hyhz201305035035 |
| 分类号: |
| 基金项目:海洋公益性行业科研专项经费, 201205024号; 山东省中青年科学家科研奖励基金, DS2010HZ001号; 国家海洋局第一海洋研究所基本科研业务费专项基金, GY02-2011G17号。 |
附件 |
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| OPTIMIZATION OF FERMENTATION CONDITION FOR AGARASE PRODUCTION OF STRAIN BACILLUS sp. BI-3 FROM INDONESIAN HOT SPRING |
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SHA Yu-Jie1, YANG Ping-Ping1, HE Pei-Qing2, LI Jiang2, XU Yuan2, ZHANG Qu2
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1.Shandong Polytechnic University;2.The First Institute of Oceanography, State Oceanic Administration
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| Abstract: |
| To study the optimal culture medium and fermentation conditions of an agarase-produced thermophilic bacterium Bacillus sp. BI-3 from Indonesia, we used response surface methodology to optimize the fermentation medium and the conditions of fermentation. Ranges of single factors (temperature, pH, carbon sources, nitrogen sources, metal ions, and inoculum) were selected for single-factor test, and then three important factors influencing agarase activity were identified in initial experimental design of Plackett-Burman by Design-Expert software, i.e., D-mannose, strontium chloride, and calcium chloride. Box-Behnken design and response surface analysis were adopted to further investigate the mutual interaction among the variables and obtain optimal condition to maximize agarase activity. The optimal culture medium and fermentation conditions of Bacillus sp. BI-3 are as follow: yeast extract 0.3%, D-mannose 0.66%, peptone 0.6%, calcium chloride 7.21mmol/L, strontium chloride 6.00mmol/L, NaCl 6‰, incubation temperature was 55℃, 1% of the inoculum size, initial pH 6.0. The maximum yield of agarase was 6.0U/mL, which is 2.4 times high of that before optimization. |
| Key words: agarase thermophilic bacterium response surface optimization |
