摘要
硫酸软骨素是一种重要的糖胺聚糖,广泛应用于医药、食品、保健品行业。为了提高硫酸软骨素类似物果糖软骨素的生产,通过过量表达磷酸葡糖胺变位酶(GlmM)、葡萄糖胺合酶(GlmS),优化融合蛋白glmM-glmS的表达水平,获得了最优工程菌株E.coli K4-H-glmM-glmS,并确定了最适IPTG浓度(0.4 mmol/L)以及诱导温度(37℃)。最后,在5-L罐水平下,借助DO-stat补料模式,果糖软骨素的产量达到了3.99 g/L,较原始菌株提高了108.9%,为工业化生产果糖软骨素奠定了基础。
Sulfated chondroitin is an important glycosaminoglycans,which is widely used in medical,food and cosmetic fields.To improve the production of fructosylated chondroitin,GlcN synthase(GlmM)and GlcN-6-P synthase(GlmS)were overexpressed,the confusion protein was constructed and a recombinant strain E.coli K4-H-glmM-glmS was obtained.Then,the optimal induction conditions including IPTG concentration and induction temperature were estalished.Finally,with the DO-stat feeding strategy in a 5 L fermenter,the yield was up to 3.99 g/L,which was 108.90%higher than that with wild type strain.The study provides a basis for industrial-scale production of fructosylated chondroitin.
作者
张权
酉相成
陈修来
刘佳
罗秋玲
刘立明
ZHANG Quan;YOU Xiangcheng;CHEN Xiulai;LIU Jia;LUO Qiuling;LIU Liming(State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;Laboratory of Food Microbial-Manufacturing Engineering,Jiangnan University,Wuxi 214122,China)
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2020年第3期71-80,共10页
Journal of Food Science and Biotechnology
基金
国家自然科学基金项目(21676118)
江南大学自主科研计划重点项目(JUSRP51611A)。
关键词
大肠杆菌K4
代谢路径
果糖软骨素
代谢工程
发酵工程
Escherichia coli K4
metabolic pathway
fructosylated chondroitin
metabolic engineering
fermentation engineeringorganic acid