Optimization of Solvent Media to Solubilize TEV Protease Using Response Surface Method

Overview

Journal Res Pharm Sci

Specialty Chemistry

Date 2020 Dec 14

PMID 33312211

Citations 1

Authors

Niloufar Mohammadian

Hossein Mohammadian

Fatemeh Moazen

Mohammad Hosein Pakdel

Ali Jahanian-Najafabadi

Hamid Mir Mohammad Sadeghi

Affiliations

Soon will be listed here.

Abstract

Background And Purpose: Tobacco etch virus (TEV) protease, a 27 KDa protein, consists of the catalytic domain of nuclear inclusion a (NIa) protein produced by virus. Because of its unique sequence, TEV protease is used for purging fusion tags from proteins. It also has many advantages such as stability and activity in a board range of temperature and pH and overproduction in and these benefits make this protease valuable. Despite all these benefits, TEV protease has problems like low solubility (less than 1 mg/mL). There are methods for enhancing protein solubility and in this study, the effect of additives during cell lysis was studied.

Experimental Approach: Eleven different additives that made twelve different lysis buffers were used and their effect on TEV protease solubility analyzed by Plackett-Burman and response surface methodology methods.

Findings / Results: Three best effective additives on TEV solubility (L-proline, sodium selenite, and CuCl2) were selected according to software analysis and the best concentration of them was applied to optimize TEV protease solubility.

Conclusion And Implications: The obtained results provided the composition of an optimum solvent for obtaining soluble TEV protease.

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