The Expression of Elastin-like Polypeptide Tag in Periplasmic Space of Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.031

Abstract

Abstract: This work aims to investigate the expression of elastin-like polypeptide[I]₄₀（ELP[I]₄₀）in the periplasmic space of Escherichia coli. The expression vectors of pIG6LH/ELP[I]₄₀and pIG6LH/ELP[I]₄₀+Trx were constructed and transfected into E. coli BLR（DE3）and induced to express by IPTG. Recombinant ELP fusion protein were purified by inverse transition cycling（ITC）. Inverse temperature transition（T_t）of ELP[I]₄₀ and ELP[I]₄₀+Trx protein were measured. The effects of the concentrations of ELP[I]₄₀ and ELP[I]₄₀+Trx proteins and NaCl on phase-transition temperature were investigated. ELP[I]₄₀ and ELP[I]₄₀+Trx protein were purified by 3 round ITC. When the protein concentration of ELP[I]₄₀and ELP[I]₄₀+Trx were 10 μmol/L,25 μmol/L,50 μmol/L,75 μmol/L,and 100 μmol/L,the T_t were 31.5℃,29℃,27℃,26℃,25℃ and 31.8℃,29.5℃,27.5℃,26℃,25.5℃ respectively. When the final concentrations of ELP[I]₄₀ and ELP[I]₄₀+Trx were 25 mmol/L respectively and the NaCl concentration was 0.25 mol/L,0.5 mol/L,0.75 mol/L,1.00 mol/L,and 1.25 mol/L respectively,the T_t of ELP[I]₄₀ decreased from 29℃ to 24.5℃,22℃,19℃,15℃,and 11.5℃ respectively and the T_t of ELP[I]₄₀+Trx from 29.5℃ to 25℃,23℃,20.2℃,15.5℃,and 11.8℃ respectively. The ELP[I]₄₀ in the periplasmic space of E. coli possessed the same physicochemical properties as the intracellular one,and the ELP could be used as an isolation and purification tag of expressed proteins in E. coli’s periplasmic space.

Key words: elastin-like polypeptide tag, thioredoxin, Escherichia coli, periplasmic space, purification

LI Jing-quan, LIN Heng, DING Ning, ZUO Xiao-xue, SHI Jin-ping, XU Yong-ping, LENG Chun-ling, YANG Chun-guang. The Expression of Elastin-like Polypeptide Tag in Periplasmic Space of Escherichia coli[J]. Biotechnology Bulletin, 2016, 32(8): 214-220.

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