产精氨酸脱亚氨基酶大肠杆菌全细胞催化合成L-瓜氨酸  

Whole Cell Catalytic Synthesis of L-Citrulline by Escherichia coli Producing Arginine Deaminase

作　　者：连战 李天罡 王松江 郭传庄 李胜涛 吕志飞 李升梅 王建彬 LIAN Zhan;LI Tiangang;WANG Songjiang;GUO Chuanzhuang;LI Shengtao;LYUZhifei;LI Shengmei;WANG Jianbin(Dongxiao Biotechnology Co.,Ltd.,Weifang 262218,China)

机构地区：[1]东晓生物科技股份有限公司,山东潍坊262218

出　　处：《食品科技》2025年第1期18-24,共7页Food Science and Technology

基　　金：国家重点研发计划项目(2023YFD1300700)。

摘　　要：为实现产精氨酸脱亚氨基酶大肠杆菌全细胞催化合成L-瓜氨酸的工业化生产,对其催化工艺进行研究。首先探讨菌体处理方式,确定不同贮存时间较适处理方式如下:<24 h,常温处理;24~36 h,冷冻处理;>36 h,冷藏处理。在上述处理方式的基础上,采取单因素法分别探究全细胞催化产L-瓜氨酸的转化p H值、温度、底物浓度、酶加量、金属离子、转化周期,并对胞内胞外酶转化的占比、全细胞酶二次转化效果进行研究。结果显示,较适催化条件为pH5.5、温度40℃、底物浓度150 g/L、加酶量2 g/L,添加10 mmol/L Mg^(2+)、0.1 mmol/L Co^(2+),周期24 h,在该工艺条件下使用10 L反应釜进行小试转化,转化率可达99.21%,较初始条件提升了5.07%,同时转化周期缩短8 h,该工艺的确定为精氨酸脱亚氨基酶全细胞催化产L-瓜氨酸的工业化应用奠定了基础。Inorder to achieve industrial production of L-citrulline by whole cell catalytic synthesis of arginine deaminase in Escherichia coli,the catalytic process was studied.Firstly,the treatment methods for bacterial cells were studied,and the suitable treatment methods for different storage times were determined as follows:<24 hours,room temperature treatment;24–36 hours,frozen treatment;>36h,refrigerated treatment.On the basis of the above processing methods,a single factor approach was adopted to investigate the conversion pH,temperature,substrate concentration,enzyme dosage,metal ions,and conversion cycle of whole cell catalytic production of L-citrulline.The proportion of intracellular and extracellular enzyme conversion and the secondary conversion effect of whole cell enzymes were also studied.The results showed that the optimal catalytic conditions were pH 5.5,temperature 40℃,substrate concentration 150 g/L,enzyme dosage 2 g/L,addition of 10 mmol/L Mg^(2+)and 0.1 mmol/L Co^(2+),with a cycle of 24 h.Under these process conditions,a 10 L reaction kettle was used for small-scale conversion,and the conversion rate reached 99.21%,an increase of 5.07%compared to the initial conditions.At the same time,the conversion cycle was shortened by 8 h.The determination of this process laid the foundation for the industrial application of arginine deaminase whole cell catalysis for L-citrulline production.

关 键 词：L-瓜氨酸 全细胞催化 精氨酸脱亚氨基酶 转化率 

分 类 号：TS201.3[轻工技术与工程—食品科学]