摘要
胶原蛋白是一种生物高分子,具有良好的生物特性,被广泛应用于食品、化妆品、生物学以及医学等领域,但以传统方法在动物中提取的胶原存在水溶性差、具有传播病毒的风险及批间一致性差等问题。以基因工程技术得到的重组胶原蛋白组分单一、安全性高和生产过程可控。在大肠杆菌中表达重组Ⅲ型人源化胶原蛋白,通过正交实验对培养基的碳氮浓度进行优化,发现最佳添加量为蛋白胨15 g/L、酵母粉7.5 g/L和甘油12 g/L,经5 L发酵罐验证菌体量增加约120%,表达量达到10.31 g/L,提高了199%。胶原蛋白纯化后进行了氮端测序和等电点表征实验。
Collagen is a kind of biopolymer with good biological characteristics,which is widely used in food,cosmetics,biology and medicine.However,the collagen extracted from animals by traditional methods has some problems,such as poor water solubility,risk of virus transmission and poor consistency between batches.The recombinant collagen obtained by genetic engineering technology has the advantages of single component,high safety and controllable production process.In this study,the gene of recombinant human typeⅢcollagen protein was heterologously expressed in Escherichia coli.Then,the carbon and nitrogen content in the medium for recombinant human typeⅢcollagen protein production was optimized by orthogonal tests.The optimal medium carbon and nitrogen content was 15 g/L peptone,7.5 g/L yeast powder and 12 g/L glycerol.The carbon and nitrogen concentrations were optimized and verified in 5 L fermentor.the amount of biomass was increased by 120%and the expression yield reached 10.31 g/L(increased by 199%).Futhermore,after purification of collagen,nitrogen sequencing and isotonic characterization experiments were carried out.
作者
张发勇
宋富
齐磊
吴飞陶
刘鑫
于月欣
李华
冯毅
邵文亮
张玮倩
徐兰举
ZHANG Fayong;SONG Fu;QI Lei;WU Feitao;LIU Xin;YU Yuexin;LI Hua;FENG Yi;SHAO Wenliang;ZHANG Weiqian;XU Lanju(Hebei Province Medicines Medical Equipment Inspection Research Institute,Shijiazhuang 050000,China;Hebei NACOL Biotechnology Co.,Ltd.,Shijiazhuang 050000,China;College of Food Science and Technology,Hebei Agricultural University,Baoding 071001,China)
出处
《发酵科技通讯》
CAS
2022年第1期36-40,共5页
Bulletin of Fermentation Science and Technology
基金
石家庄市引进国外智力基金项目(20210013)。
关键词
大肠杆菌
重组胶原蛋白
培养基优化
工艺放大
Escherichia coli
recombinant collagen
optimization of culture medium
process amplification
