Optimization of culture medium for alginate lyase by Photosbacterium phosphoreum
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摘要: 通过响应面方法对明亮发光杆菌液体发酵产褐藻胶裂解酶的最佳产酶培养基进行了优化。在单因素实验的基础上,通过部分因子实验对葡萄糖、牛肉膏、氯化钾、七水硫酸亚铁、磷酸氢二钾、MgSO4·7H2O、初始pH进行系统考察,筛选出两个影响较显著的因素MgSO4·7H2O和葡萄糖添加量,然后通过中心组合实验进一步优化,建立以褐藻胶裂解酶酶活力为响应值的二次回归方程模型,获得了最优的发酵培养基组成（g/100 mL）:葡萄糖1.211 g、牛肉膏0.2 g、氯化钾0.5 g、七水硫酸亚铁0.05 g、磷酸氢二钾0.02 g、七水硫酸镁0.005 g,初始p H8.6,在该优化的培养条件下,褐藻胶裂解酶酶活为69.05 U/mL。与基础培养基比,褐藻胶裂解酶产酶量提高了14.5倍,说明本优化工艺具有可行性。Abstract: In order to enhance the production of alginate lyase by Photosbacterium phosphoreum, response surface methodology was applied to optimize the culture medium. Based on the single factor experiment, a fractional factorial design was used to investigate the main factors affecting alginate lyase yield. The results showed that two factors played important roles in the production of alginate lyase.Central composite design and response surface analysis were applied to establish a second-order regression equation model for the yield of alginate lyase. At last, the optimal fermentation medium were obtained by response surface analysis as follows: glucose 1.211 g, beef extract 0.2 g, KCl 0.5 g, FeSO4·7H2O 0.05 g, K2HPO4 0.02 g, MgSO4·7H2O 0.005 g, initial pH8.6.Under the optimal conditions, the activity of alginate lyase reached 69.05 U/m L.By optimization of culture medium, production of alginate lyase was improved 14.5 times compared to basic culture medium.In conclusion, the optimized process has good reliability.
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