Quá trình biểu hiện các gen đích của vi sinh vật trong tế bào Escherichia coli vẫn được sử dụng rộng rãi trong các nghiên cứu trên thế giới do các ưu điểm của hệ thống này như lượng sinh khối lớn, tốc độ biểu hiện lớn, điều kiện biểu hiện tương đối đơn giản và dễ kiểm soát. | Một số điều kiện ảnh hưởng đến quá trình biểu hiện gen mã hóa Endoglucanase từ DNA metagenome của suối nước nóng Bình Châu trong Escherichia coli TAP CHI SINH HOC 2019, 41(2): 111–118 DOI: THE EFFECT OF SOME FACTORS ON EXPRESSION OF GENE ENCODING ENDOGLUCANASE FROM DNA METAGENOME OF BINH CHAU HOT SPRING IN Escherichia coli Tran Thanh Thuy, Lai Thi Hong Nhung, Tran Dinh Man, Le Thi Thanh Xuan, Nguyen Kim Thoa* Institute of Biotechnology, VAST, Vietnam Received 3 April 2019, accepted 28 May 2019 ABSTRACT Expression of microbial target genes in Escherichia coli is broadly used due to its advantages namely: well established system, easy to manipulate, a huge biomass, high level productivity, safe and inexpensive to grow. Metagenomic technique has been applying in Vietnam recently for effective mining of uncultured gene resources, especially in endemic mini-ecologies such as hot springs where the cell densities are low. DNA metagenome of Binh Chau hot spring was isolated and sequenced by Illumia HiseqTM. Based on analyses of databases of cellulase-encoded genes, denovogenes 18736 gene sequence for thermal endoglucanase was selected for expression in E. coli. In this paper, some factors for expression of endoglucanase have been investigated. The results show that appropriate gene expression conditions are: Expression performed in E. coli C43 (DE3) on TB medium at 30oC with mM of IPTG as inducer, the culture volume of 20% compared with the bottle volume and the expression time is 42–48 hours. In this condition, the biomass production and soluble enzyme activity can reached up to – g /L and – U/mL, respectively. Our results show the prospect of exploiting microbial genes without culture. Keywords: Binh Chau hot spring, DNA-metagenomic, E. coli C43(DE3), gene expression, recombinant endoglucanase, thermostable enzyme. Citation: Tran Thanh Thuy, Lai Thi
