高产纤溶酶菌株CNY16发酵条件优化及其酶学特性初步研究

Optimization of fermentation conditions of a fibrinolytic enzyme stain CNY16 with high activity and preliminary study of its enzymatic propertie

摘要:

【目的】通过响应面试验对产纤溶酶菌株CNY16发酵条件进行优化，并对其酶学特性进行初步研究。【方法】采用Plackett-Burman设计得出酵母膏、氯化钠、转速3个最重要影响因素，通过最陡爬坡实验逼近酶活的最高区域，然后根据Box-Behnken中心组合设计实验对显著因素进行优化分析，最后对该酶学性质进行初步分析。【结果】最终得到3个因素的最优组合：酵母膏3.28%，氯化钠1.14%，转速166 r/min，在此培养条件下，纤溶酶活达到875.932 U/mL，比优化前提高了46%；该菌株产纤溶酶最适温度为30 °C，最适pH为6.5。【结论】确定了高产纤溶酶菌株CNY16的最优发酵条件及其部分酶学性质，为该酶的进一步深入研究及中试实验奠定基础。

Abstract:

[Objective] Response surface analysis was applied to optimize the fermentation conditions for producing fibrinolytic enzyme strain CNY16, and preliminarily studied its enzymology characteristics. [Methods] Through the Plackett-Burman design, the most significant effect on fibrinolytic enzyme production was obtained which include yeast extract, sodium chloride and revolving speed. The highest fibrinolytic activity area was investigated by the steepest climbing experiments, then, the optimal fermentation conditions of producing fibrinolytic enzyme strain CNY16 was analysised by Box-Behnken central composite design experiments. [Results] The optimal fermentation conditions of strain CNY16 was determined for 3.28% yeast extract, 1.14% sodium chloride, rotate speed at 166 r/min, the enzyme activity was 875.932 U/mL, the optimized enzyme activity was 46% higher than before. The optimum temperature for the fibrinolytic enzyme was 30 °C and the optimum pH was 6.5. [Conclusion] This experiment determined the most optimal fermentation conditions for producing fibrinolytic enzyme strain CNY16 and researched part of enzymatic properties, laied a foundation for further research and experiment of the the pilot study.