亚栖热菌产海藻糖合酶培养基的优化

Medium Optimization for the Production of Trehalose Synthase by Meiothermus sp.CBS-01

对亚栖热菌 CBS-01菌株发酵生产海藻糖合酶的培养基进行了优化.首先通过碳源、氮源实验,选出了适宜的碳、氮源,随后采用 Plackett-Burman 设计法对培养基中的组分进行了筛选,找出影响产酶的主要因素为蛋白胨和酵母膏,最后利用中心组合设计及响应面分析法求出了主要影响因素的最佳浓度,得到优化的发酵培养基配方(g/L):可溶性淀粉5,蛋白胨8.6,酵母膏1.725,NaNO_3 0.7,氨三乙酸0.5,Na_2HPO_4 0.1,MgSO_4·7H_2O 0.2,CaCl_2 0.1.10 L 罐发酵实验表明,CBS-01菌株海藻糖合酶的合成属生长关联型,利用优化的发酵培养基,CBS-01菌株的产酶水平由0.2 U/mL 提高到0.41 U/m L.

Optimization of the fermentation tion by a new strain, Meiothermus sp. CBS-01, medium for maxium trehalose synthase producwas carried out. Single factor experiment was used to choose proper carbon and nitrogen source. The main factors among components of the medium influencing trehalose synthase production were selected by Plackett-Burman experiment design. Then central composite design and response surface methodology were used to optimize the main factors selected. The results showed that trehalose synthase production was growth-associated. Peptone and yeast were two main factors affecting the enzyme production. The optimum medium composition consisted of 5 g/L soluble starch, 8.6 g/L peptone, 1. 725 g/L yeast, 0.7 g/L NaNOs, 0.5 g/L nitrilotriacetic acid, 0. 1 g/L Na2HPO4,0.2 g/L MgSO4 · 7H2O,0.1 g/L CaCl2. When applying the optimized medium, the maximum trehalose synthase production was increased from 0.2 U/mL with non-optimized medium to 0.41 U/mL in a 10-L jar fermentor. It is demonstrated that Plackett-Burman design and response surface methodology are effective for optimization of fermentation medium.