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

Tran Thi Hong, Ton That Huu Dat, Nguyen Phuong Hoa, Pham Viet Cuong, Nguyen Thi Kim Cuc

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 Da Nang to express in Escherichia coli BL21(DE3). The gene PI-DN9 encoding protease inhibitor (1.3 kb) was cut off cloning vector pUC57/PI-DN9 containing gene PI-DN9 and inserted into expression vector pET32a(+), the recombinant vector pET32a(+)/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.

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DOI: https://doi.org/10.15625/2615-9023/v40n4.12759 Display counter: Abstract : 137 views. PDF : 61 views.

 

                 

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