响应面法优化假单胞菌HFE733产脂肪酶发酵培养基的研究

Study on Optimization of Culture Medium for Lipase Production by Pseudomonas sp. HFE733 Using Response Surface Methodology

研究采用响应面法对假单胞菌HFE733产脂肪酶的发酵培养基进行了优化。在前期运用单因素试验优化的基础上,利用Plackett-Burman试验设计筛选出培养基组分中影响脂肪酶活力的2个显著性因素:酵母膏和橄榄油。运用单因素试验研究非显著因素的最低添加量来降低生产成本,然后运用最陡爬坡试验找到中心复合试验的中心点,最后利用中心复合设计确定显著性因素之间的交互作用及最佳组成。结果表明:最佳培养基组分为酵母膏34g/L、橄榄油5.92 mL/L、蔗糖12.5g/L、CuSO_4·5H_2O 0.4g/L、MnSO_4·H_2O 0.2g/L,在此条件下脂肪酶酶活可达到9.27U/mL,与模型预测值相近,且与单因素优化后的酶活(4.53U/mL)相比,提高了104.64%。

In this study, response surface methodology （RSM） is employed to optimize the fermentation culture medium for lipase production by Pseudomonas sp. HFE733 in submerged fermentation. Based on single factor experiments, two significant influence factors are screened by the method of Plackett-Burman design as： yeast extract and olive oil. To reduce the production cost, the minimum additive amount experiment is used to find out the minimum additive amount of non-significant factors, and the steepest ascent design is applied to obtain the center point of central composite design, and the optimal concentration and correlations of two factors are identified by central composite design. The result indicates that the optimal fermentation components for lipase production are as follows= yeast extract is 34 g/L, olive oil is 5.92 mL/L, sucrose is 12.5 g/L, CuSO4 ·5H2O is 0. 4 g/L, MnSO4 · H2O is 0. 2 g/L. The enzyme activity in verification test reaches 9.27 U/mL, consistent with the model predicted value, which is 104.64 hi＇gher than that of single factor experiments＂ results （4.53 U/mL）.