Yan Cheng, Zhen Wan; Xiaohong Sun, Vol. 4, No. 3, pp. 1-6, Sep. 2022
10.22662/IJASC.2022.4.3.001, Full Text:
Keywords: Protein Purification, HSD17B13, His tag, Gst tag, Wash Buffer.

Abstract

Background/Objectives: The Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease that affects nearly a quarter of the world population. Recent studies have shown that there is a close relationship between liver lipid-titrated 17β Hydroxysteroid Dehydrogenase Type 13（HSD17B13）protein and NAFLD. At the same time, the purification of HSD has been trying to find better conditions and methods, at present, the HSD protein separation and purification technology is incomplete, the cost is high, the purity is low, the purification quantity is small and so on. This paper first reviewed the research progress of Steatosis and some basic properties about HSD, such as HSD expression site, Lipid titration function, in vitro catalytic substrates, and the relationship between HSD and liver diseases.. Methods/Statistical analysis: The optimization of HSD purification method is considered be of great help to the further study of its structure and function, so this research put forward two hypotheses to explore its effect on the purification of HSD. The first is to change the kind of tag when expressing plasmid in E. coli, so as to change the adsorption degree of protein with corresponding group and beads, and the second is to change the concentration of different components in Wash Buffer, the aim is to find the best concentration for removing foreign protein and maintaining the adsorption capacity of HSD. Findings: The purity of HSD is closely related to tag species and Wash Buffer concentration in different experimental groups, his-Binding Buffer with 0.5% sodium deoxycholate powder added and Wash Buffer with Tris-HCl 50 mM, NACL 300 mM, Imidazole 40 mM for each component are used. Improvements/Applications: After the better purification effect, the higher purity HSD can pave the way for the future study of its potential substrate and mechanism of action.

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