葡萄糖酸钠流加工艺促进红球菌腈水合酶发酵表达  

Promoting the Fermentation Expression of Nitrile Hydratase in Rhodococcus rhodochrous by the Fed Batch Process of Sodium Gluconate

作　　者：张国辉 刘禹轩 刘燕枝 谢嘉怡 栗波 张思楠 王泽建[1] 吴诗勇 ZHANG Guohui;LIU Yuxuan;LIU Yanzhi;XIE Jiayi;LI Bo;ZHANG Sinan;WANG Zejian;WU Shiyong(State Key Laboratory of Bioreactor Engineering;State Key Laboratory of Coal Liquification,Gasification and Utilization with High Efficiency and Low Carbon Technology;Engineering Research Center of Resource Utilization of Carbon-Containing Waste with Carbon Neutrality,Ministry of Education,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学1.生物反应器工程国家重点实验室 [2]煤液化气化及高效低碳利用全国重点实验室 [3]含碳废弃物资源化利用与碳中和教育部工程研究中心,上海200237

出　　处：《华东理工大学学报(自然科学版)》2025年第2期209-218,共10页Journal of East China University of Science and Technology

基　　金：国家重大专项项目资助项目(2020YFA0906800);国家自然科学基金(32071471)。

摘　　要：首先,考察了不同质量浓度的葡萄糖和氯化钴对腈水合酶(NHase)合成的影响,结果表明当葡萄糖质量浓度为20 g/L、氯化钴质量浓度为0.050 g/L时,NHase的表达水平得到了显著提升;其次,建立了基于发酵过程生理代谢参数采集与分析系统,通过多参数相关性分析得出pH是影响红球菌NHase表达的关键因素,从而提出了发酵过程中流加葡萄糖酸钠控制pH的策略。结果表明,与分批发酵工艺相比,优化后的工艺在菌体浓度相同的情况下,总酶活提高了93.74%,比酶活提高了97.35%,糖转化率提高了71.36%。本研究结果为工业生产中提高NHase产量提供了理论指导。Nitrile hydratase(NHase)is one of the key enzymes in nitrile catabolism,which can catalyze the conversion of aliphatic nitriles or aromatic nitriles to the corresponding amides.Currently,NHase is used in the industrial production of acrylamide(AM)and niacinamide through the hydration of acrylonitrile(AN)and 3-cyanopyridine,respectively.Rhodococcus has been widely used in the biotransformation of AN due to their excellent properties such as high NHase activity,heat resistance and good stability in organic solvents.In this study,the effects of different concentrations of glucose and cobalt chloride on the synthesis of nitrile hydratase(NHase)were investigated.The results showed that the glucose mass concentration of 20 g/L and the cobalt chloride concentration of 0.050 g/L could effectively enhance the synthesis of NHase.Subsequently,a process physiology metabolic parameter collection and analysis system based on fermentation process off-gas mass spectrometer were established.Through multi-parameter correlation analysis,pH was identified as a key factor influencing the expression of NHase in Rhodococcus rhodochrous J1,and thus a strategy of continuous adding sodium gluconate during the fermentation process to control the pH was proposed.Compared to the original batch fermentation,the optimized process exhibited a total enzyme activity of NHase increased by 93.74%,the specific enzyme activity increased by 97.35%and the sugar conversion rate increased by 71.36%in continuous batch fermentation under the same bacterial concentration.This paper provides theoretical guidance for enhancing acrylonitrile hydratase production in industrial settings.

关 键 词：腈水合酶 红球菌 多参数相关性分析 葡萄糖酸钠 发酵 

分 类 号：Q815[生物学—生物工程]