Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam

In this study, we selected a novel gene encoding protease inhibitor from metagenome of marine sponge QT collected in Quang Tri and expressed in the Escherichia coli (E. coli) strain BL21(DE3) for finding and mining novel protease inhibitor. | Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam ACADEMIA JOURNAL OF BIOLOGY 2018, 40(4): 77–84 DOI: EXPRESSION OF A NOVEL GENE ENCODING PROTEASE INHIBITOR FROM METAGENOME OF SPONGE IN VIETNAM Tran Thi Hong1, 3*, Ton That Huu Dat1, Nguyen Phuong Hoa1, Pham Viet Cuong1, Nguyen Thi Kim Cuc2 1 Mien Trung Institute for Scientific Research, VAST 2 Institute of Marine Biochemistry, VAST 3 Graduate University of Science and Technology, VAST ABSTRACT Marine sponge is known as a “gold mine” of natural products from marine environment. Many novel bioactive compounds have been isolated from marine sponges and sponge-associated microorganisms such as antibiotics, anti-cancer compounds, protease inhibitors, etc. In this study, we selected a gene encoding protease inhibitor from metagenome of a sponge collected in Quang Tri to express in Escherichia coli (E. coli) BL21(DE3). The gene PI-DN9 encoding protease inhibitor ( kb) was cut off cloning vector pUC57/PI-DN9 containing gene PI-DN9 and inserted into expression vector pET-32a(+), the recombinant vector pET-32a(+)/PI-DN9 then was transformed and expressed in the E. coli strain BL21(DE3). Results showed that recombinant protein (50 kDa) was expressed successfully at 25°C, 1 mM of IPTG in 5 hours. The recombinant protein was purified using Ni-NTA affinity chromatography column. Western blot assay and bioactive assay showed good activity of the purified protein. Keywords: Escherichia coli, expression vector, protease inhibitor, recombinant protein, sponge. Citation: Tran Thi Hong, Ton That Huu Dat, Nguyen Phuong Hoa, Pham Viet Cuong, Nguyen Thi Kim Cuc, 2018. Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam. Academia Journal of Biology, 40(4): 77–84. . *Corresponding author email: .