Abstract: 【Objective】 he aim of this experiment is to optimize the main nutrient components of the fermentation medium of dihydrodaidzein producing strain by single factor test design and response surface methodology.【Method】 On the basic of BHI medium, the main factors (carbon source, nitrogen source, growth factor and inorganic salt) and the appropriate range of concentration were determined by single factor test. The best composition in the medium was analyzed by the Box-Behnken central composite experiment and response surface methodology.【Result】 The significant factors such as glucose, peptone,VB₁ and KH₂PO₄ were determined by the single factor experiment and response surface methodology analysis was determined that the supplemental doses to the strain medium were 10.22 g/L glucose, 6.00 g/L peptone, 0.49 g/L VB₁ and 0.82 g/L KH₂PO₄ on the basic of BHI medium. Validation experiments showed that the DHD concentration was 0.27 μg/mL, which was close to the predictive value 0.28 μg/mL.【Conclusion】 The results from the single factor experiment and response surface methodology showed that the optimal ingredients of medium for DHD production were 38 g/L BHI,10.22 g/L glucose,6.00 g/L peptone,0.49 g/L VB₁ and 0.82 g/L KH₂PO₄.

Key words: dihydrodaidzein; response surface methodology; BHI medium

摘要： 【目的】 研究采用单因素试验设计和响应面分析法对产二氢大豆苷元菌株发酵培养基的主要营养成分进行优化。【方法】 以脑浸心肉汤培养基(BHI)为基础,通过单因素试验确定主要因素(碳源、氮源、生长因子和无机盐)及其适宜的浓度范围,利用Box-Behnken中心组合试验设计,采用响应面法进行回归分析,确定培养基中的最佳组成成分。【结果】 通过单因素试验确定了葡萄糖、蛋白胨、VB₁和KH₂PO₄能够提高二氢大豆苷元的产量,响应面分析法进一步确定在以BHI为基础的条件下,葡萄糖的补充量为10.22 g/L、蛋白胨为6.00 g/L、VB₁为0.49 g/L、KH₂PO₄为0.82 g/L。验证试验显示在培养基优化后的条件下,DHD的产生量为0.27 μg/mL,与预测值0.28 μg/mL接近。【结论】 利用单因素试验和响应面法,分析明确了产二氢大豆苷元菌株发酵培养基最佳成分为38 g/L的BHI、10.22 g/L的葡萄糖、6.00 g/L的蛋白胨、0.49 g/L的VB₁、0.82 g/L的KH₂PO₄。

关键词: 二氢大豆苷元, 响应面法, BHI培养基