响应面法优化重组大肠杆菌全细胞催化合成丙谷二肽  

Optimization of whole cell catalytic synthesis of Ala-Gln via recombinant E.coli by response surface methodology

作　　者：上官丁从 范玉娜 郭冯喆 李琪 杨静文 张洪斌 胡雪芹[1] SHANGGUAN Dingcong;FAN Yuna;GUO Fengzhe;LI Qi;YANG Jingwen;ZHANG Hongbin;HU Xueqin(School of Food and Biological Engineering,Hefei University of Technology,Hefei 230009)

机构地区：[1]合肥工业大学食品与生物工程学院,合肥230009

出　　处：《中国食品添加剂》2021年第9期58-65,共8页China Food Additives

摘　　要：L-丙氨酰-L-谷氨酰胺(L-alanyl-L-glutamine,丙谷二肽)是重要的肠外营养补充剂,有着广泛的应用。本文利用表达α-氨基酸酯酰基转移酶的重组大肠杆菌QC01,进行全细胞催化法合成丙谷二肽。通过最陡爬坡实验筛选出对全细胞催化影响最显著的三个因素:QC01细胞浓度、底物比例和反应温度,再利用响应面实验,研究了这三个因素的交互作用。最终确定全细胞催化条件为:QC01细胞浓度3g/L、底物丙氨酸甲酯盐酸盐与谷氨酰胺的摩尔比为1∶0.6、反应温度30℃、底物浓度(以丙氨酸甲酯为准)300mmol/L、反应起始pH9.0、反应时间为40min,优化后底物转化率从22.75%提高到73.85%,为全细胞法制备丙谷二肽的工业化生产提供借鉴。L-alanyl-L-glutamine(Ala-Gln)is an important nutritional supplement and has a wide range of applications.In this paper,Ala-Gln was synthesized by whole cell catalysis of recombinant Escherichia coli QC01 expressingα-amino acid ester acyltransferase.Through steepest ascent experiment,three factors which had the most significant effect on whole cell catalysis were selected:QC01 cell concentration,substrate ratio and reaction temperature.The interaction of these three factors was studied by response surface methodology.Finally,the whole cell catalytic conditions were determined:QC01 cell concentration 3 g/L,substrate ratio(methyl propionate hydrochloride:glutamine)1∶0.6,reaction temperature 30℃,substrate concentration 300 mmol/L,initial pH9.0,reaction time 40 min.The conversion rate was increased from 22.75%to 73.85%under the above conditions.The study can be used as a reference for the industrial production of Ala-Gln by whole cell catalysis.

关 键 词：丙谷二肽 全细胞催化 响应面 催化条件优化 

分 类 号：TS202.1[轻工技术与工程—食品科学] Q814[轻工技术与工程—食品科学与工程]