食品与发酵工业2023，Vol.49Issue(23)：33-40,8.DOI:10.13995/j.cnki.11-1802/ts.035136
肌酐酶在大肠杆菌中外源表达及发酵优化
Exogenous expression and fermentation optimization of creatininase in Escherichia coli
摘要
Abstract
Creatininase,an important tool enzyme for in vitro detection reagents,is currently produced mainly by microbiological methods,which suffer from low yield and poor activity,limiting its practical application.Herein,recombinant Escherichia coli BL21(DE3)/pET28a-CA was constructed by whole gene synthesis and heterologous expression,obtaining high-performance creatininase.The optimal medium composition was determined by single-factor test and response surface design analysis,and the fermentation conditions were systematically optimized,resulting in improved creatininase expression.The medium optimization results showed that the optimal me-dium consisted of glucose 10.33 g/L,yeast extract 21.80 g/L,phosphate 137.63 mmol/L,tryptone 15.60 g/L.After optimization of the fermentation process in 5 L fermenters,the enzyme activity and specific enzyme activity of creatinine was greatly improved and reached 470.24 U/mL and 367.38 U/mg,respectively,under the conditions of 4％inoculum,25 ℃ induction temperature,18 h induction time,0.2 mmol/L IPTG and 30％dissolved oxygen value.This study provides a theoretical basis and technical support for the production and industrial application of creatininase.关键词
肌酐酶/外源表达/发酵优化/大肠杆菌/响应面法Key words
creatininase/heterologous expression/fermentation optimization/Escherichia coli/response surface methodology引用本文复制引用
周丽亚,张悦,刘瑞琪,陈佳立,刘运亭,姜艳军,马丽..肌酐酶在大肠杆菌中外源表达及发酵优化[J].食品与发酵工业,2023,49(23):33-40,8.基金项目
河北省省级科技计划项目(21372805D,21372804D,20372802D) （21372805D,21372804D,20372802D）
国家自然科学基金项目(21878068,21576068) （21878068,21576068）
河北省自然科学基金项目(B2020202036) （B2020202036）
