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

Authors

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

DOI:

https://doi.org/10.15625/2615-9023/v40n4.12759

Keywords:

Escherichia coli, expression vector, protease inhibitor, recombinant protein, sponge.

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.

 

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Published

31-12-2019

How to Cite

Hong, T. T., Dat, T. T. H., Hoa, N. P., Cuong, P. V., & Cuc, N. T. K. (2019). Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam. Academia Journal of Biology, 40(4). https://doi.org/10.15625/2615-9023/v40n4.12759

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