Literature summary for 3.4.21.9 extracted from

Simeonov, P.; Berger-Hoffmann, R.; Hoffmann, R.; Straeter, N.; Zuchner, T.
Surface supercharged human enteropeptidase light chain shows improved solubility and refolding yield (2011), Protein Eng. Des. Sel., 24, 261-268.

Application

Application	Comment	Organism
biotechnology	enteropeptidase is a serine protease used in different biotechnological applications. For many applications the smaller light chain can be used to avoid the expression of the rather large holoenzyme	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of enteropeptidase light chain as thioredoxin-fusion protein in Escherichia coli strain BL21(DE3) in aggregated form, subcloning in Escherichia coli strain DH5alpha	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
C112S	site-directed mutagenesis, replacement of the free cysteine residue with serine improves the refolding yield of the recombinant enzyme by 50%. The heat stability of this C112S variant was also significantly improved by supercharging	Homo sapiens
additional information	even mild protein surface supercharging by mutagenesis can have pronounced effects on protein solubility and stability, overview	Homo sapiens
N6D/G21D/G22D/N141D/K209E	site-directed mutagenesis, the mutations lead to supercharging of the protein surface leading to 100fold increased protein solubility	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P98073	-	-

Purification (Commentary)

Purification (Comment)	Organism
refolded, soluble, and detagged recombinant enteropeptidase light chain by affinity chromatography on soybean trypsin inhibitor agarose	Homo sapiens

Renatured (Commentary)

Renatured (Comment)	Organism
recombinant expression of thioredoxin-fusion enteropeptidase light chain from aggregates in 3 M guanidine-HCl, pH 8.0, in Escherichia coli strain BL21(DE3) by dilution of the protein in 0.7 M arginine-HCl, pH 8.5, 15% v/v glycerol, 3 mM reduced glutathione, and 1 mM EDTA. After 72 h at 4°C the refolding solution is dialyzed for 8 h against 50 mM Tris-HCl, pH 8.0, to facilitate complete autocatalytic activation by cleaving the fusion tag	Homo sapiens

Renatured (Comment)

Organism

recombinant expression of thioredoxin-fusion enteropeptidase light chain from aggregates in 3 M guanidine-HCl, pH 8.0, in Escherichia coli strain BL21(DE3) by dilution of the protein in 0.7 M arginine-HCl, pH 8.5, 15% v/v glycerol, 3 mM reduced glutathione, and 1 mM EDTA. After 72 h at 4°C the refolding solution is dialyzed for 8 h against 50 mM Tris-HCl, pH 8.0, to facilitate complete autocatalytic activation by cleaving the fusion tag

Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
commercial preparation	-	Homo sapiens	-

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
60	-	purified recombinant enteropeptidase light chain wild-type and N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chains show 60% remaining activity	Homo sapiens
65	-	purified recombinant enteropeptidase light chain wild-type and mutant C112S show 10% remaining activity, the purified recombinant N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chain shows 20% activity remaining	Homo sapiens
70	-	purified recombinant enteropeptidase light chain wild-type and mutant C112S are inactivated, purified recombinant N6D/G21D/G22D/N141D/K209E mutant enteropeptidase light chain shows 20% activity remaining	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Homo sapiens