Cloning, Expression, and Functional Characterization of Autoactivated Human Prethrombin-2 Synthetic Gene by Using Pichia pastoris SMD1168 As a Host

Subroto, Toto; Pertiwi, Wulan; Fadhillah, Muhammad; Hasan, Khomaini; Budiantoro, Ogi; Enus, Sutarya; Soemitro, Soetijoso

doi:10.5454/mi.10.2.1

Cited by 2 publications

(1 citation statement)

References 17 publications

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“…Prethrombin-2, structurally has one glycosylation site (at Asn373) and four disulphide bonds (Cys293-Cys439, Cys348-Cys364, Cys493-Cys507, and Cys521-Cys551) (Soejima et al, 2001). Pretrombin-2 can be converted to thrombin by FXa enzyme activators (Owen et al 1974), or with ecarin, namely metalloprotease in the venom of snake Echis carinatus species (So et al 1992;DiBella et al 1995;Jonebring et al 2012;Yonemura et al 2004;Subroto et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Temperature effect on expression of recombinant human prethrombin-2 in Escherichia coli BL21(DE3) ArcticExpress

Silaban

Simorangkir

Gaffar

et al. 2019

JPKim

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Many proteins produced in E. coli accumulate in inclusion bodies. This study aims to detect the role of temperature in reducing the formation of inclusion bodies during recombinant human prethrombin-2expressed in E. coli BL21 (DE3) Arctic Express host. In this study, we created a host growth curve to find out the right time to add an inducer. The inducer used in our experiment was IPTG 0.1 mM. The fermentation process use a temperature of 12°C and 22°C. The results showed that recombinant human prethrombin-2 was successfully expressed as protein soluble using an optimum temperature of 12°C in E. coli BL21 (DE3) Arctic Express. It was indicated from the 63kDa protein band obtained from the soluble fraction on SDS-PAGE. The higher temperature of fermentation increased the amount of protein in the insoluble fraction due. It concluded that the fermentation temperature affect the rate of prethrombin-2 expression.

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