Analysis of Methods to Improve the Solubility of Recombinant Bovine Sex Determining Region Y Protein

摘要

Background:Inclusion body formation in E. coli is a significant problem in recombinant protein production.The aim of this study was to improve the solubility of recombinant bovine sex determining region Y protein(SRY) in BL21 (DE3) E. coli cells.Methods:In this research two recombinant bovine SRY (rbSRY) sequences were analyzed; these were wildtypeSRY (wtbSRY) and codon-optimized SRY (cobSRY). Their expression in various culture conditionswas examined; these differences included IPTG concentrations, temperatures, and media stabilizers.Results:IPTG and temperature significantly affected rbSRY solubility (P 0.001). The optimum IPTGconcentration and temperatures for wtbSRY and cobSRY induction were 0.3 mM at 27 and 32 °C,respectively. In addition, arginine and sorbitol concentrations significantly affected rbSRY solubility (P 0.01). Solubility of rbSRY protein was highest from the cobSRY construct in the presence 0.2 M arginine and0.3 M sorbitol. The highest inclusion body production occurred with high glucose concentrations.Conclusions: We found that modifications in temperature and IPTG and stabilizer concentrations affectedrbSRY solubility.