High level expression of human enteropeptidase light chain in Pichia pastoris

Electrochemical determination of basic biochemical properties of enzyme enterokinase

“…Enzyme enterokinase was expressed in Pichia pastoris and purified by several chromatographic methods as described in previous paper [7]. The synthetic peptide substrate GlyAsp-Asp-Asp-Asp-Lys-b-naphthylamine (GD4K-NA) was obtained from Sigma.…”

“…Light chain is the catalytic subunit of enteropeptidase and contains domain similar to one of the serine protease chymotrypsine. Enterokinase's high specificity rate makes it the enzyme of choice for cleavage of fusion proteins produced in bacteria [5][6][7].…”

Efficient Expression and Purification of Recombinant Human Enteropeptidase Light Chain in Esherichia coli

“…Among the previous reports, the highest activity of recombinant bEK L was reported by Peng et al (2004), reaching 4.5 times that of EKMax TM . The expression and purification of hEK L was achieved in Pichia pastoris with 3.8 mg/L, and the enzyme activity was three times that of EKMax TM (Pepeliaev et al 2011). hEK L was also expressed as inclusion bodies in E. coli and 10 mg purified enzyme with five times activity of EKMax TM was obtained from 1 L cell culture after denaturation, renaturation and purification (Gasparian et al 2003).…”

“…However, there is lack of study on the recombinant human enterokinase light chain (Gasparian et al 2006). The human enterokinase light hEK L gene has been expressed in yeast cells (Pepeliaev et al 2011). However, expression was very low.…”

Heterogeneous catalysis on the phage surface: Display of active human enteropeptidase

“…It has been shown that the recombinant L-BEP activates trypsinogen mush less effectively than the native heterodimeric enzyme but is superior to it in the ability to cleave artificial fusion proteins containing the D 4 K recognition sequence [11]. The L-HEP has been successfully expressed in E. coli [22,23] and the methylotrophic yeast P. pastoris [24]. Since disulfide bridges are essential for correct folding, E. coli may produce a misfolded protein that is usually inactive or insoluble [25].…”