Soluble Production of CMP-Neu5Ac Synthetase by Co-expression of Chaperone Proteins in Escherichia coli
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CMP-Neu5Ac synthetase is a key enzyme for the synthesis of CMP-Neu5Ac, which is an essential precursor of sialylated glycoconjugates. For the soluble expression of the CMP-Neu5Ac synthetase gene (neuA) from Escherichia coli K1, various heat shock proteins were coexpressed in E. coli BL21 (DE3) Star. In order to do this, a pG-KJE8 plasmid encoding genes for GroEL-ES and DnaK-DnaJ-GrpE was co-transformed with neuA and was expressed at 20°C by addition of 0.01 mM IPTG and 0.005 mg/mL L-arabinose. The coexpression of various heat shock proteins resulted in remarkably improved production of soluble CMP-Neu5Ac synthetase in E. coli.
Abstract
CMP-Neu5Ac synthetase is a key enzyme for the synthesis of CMP-Neu5Ac, which is an essential precursor of sialylated glycoconjugates. For the soluble expression of the CMP-Neu5Ac synthetase gene (neuA) from Escherichia coli K1, various heat shock proteins were coexpressed in E. coli BL21 (DE3) Star. In order to do this, a pG-KJE8 plasmid encoding genes for GroEL-ES and DnaK-DnaJ-GrpE was co-transformed with neuA and was expressed at 20°C by addition of 0.01 mM IPTG and 0.005 mg/mL L-arabinose. The coexpression of various heat shock proteins resulted in remarkably improved production of soluble CMP-Neu5Ac synthetase in E. coli.
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