Literature summary for 1.13.12.6 extracted from

Inouye, S.; Sahara, Y.
Soluble protein expression in E. coli cells using IgG-binding domain of protein A as a solubilizing partner in the cold induced system (2008), Biochem. Biophys. Res. Commun., 376, 448-453.

Search for more literature for 1.13.12.6

Application

Application	Comment	Organism
molecular biology	construction of a cold-induced expression vector (pCold-ZZ-VL vector) in Escherichia coli cells that results in soluble bioluminescent fusion enzyme with binding ability to monoclonal antibodies, however, the Cypridina luciferase fusion enzyme is soluble but not bioluminescent (in contrast to other luciferases fused to the ZZ-domain of Staphylococcuss aureus protein A)	Cypridina noctiluca

Cloned(Commentary)

Cloned (Comment)	Organism
Staphylococcus aureus ZZ-domain gene of protein A is amplified by PCR using pEZZ18 plasmid, the gene for Cypridina luciferase (obtained by PCR) is fused to the C-terminus of the ZZ-domain in the vector to give the expression vector pCold-ZZ-VL, the vector is then expressed in Escherichia coli BL21 grown in Luria-Bertani broth at 37°C	Cypridina noctiluca

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of a cold-induced expression vector (pCold-ZZ-VL vector) in Escherichia coli cells that consists of a histidine tag sequence for nickel chelate affinity purification, IgG-binding domain of Staphylococcuss aureus protein A (ZZ-domain) and multiple cloning sites, fusing Cypridina luciferase to the ZZ-domain results in expression by Escherichia coli of a soluble cytoplasm enzyme but it is not bioluminescent (in contrast to other luciferases fused to the ZZ-domain)	Cypridina noctiluca

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
Cypridina luciferin + O2	Cypridina noctiluca	fusion enzyme + 10 mM luciferin (dissolved in ethanol in 30 mM Tris-HCl (pH 7.6) with 10 mM EDTA) with ATP (250 mM), CoA (250 microM), and MgCl2 (5 mM) in 100 mM Tris-HCl, pH 7.8	?	Cypridina luciferase fusion enzyme expressed in Escherichia coli is soluble but does not exhibit bioluminescence	?

Organism

Organism	UniProt	Comment	Textmining
Cypridina noctiluca	-	sea-firefly	-

Purification (Commentary)

Purification (Comment)	Organism
Escherichia coli cell culture with expression vector is cooled on ice-water bath, expression induced, incubation for 18 h at 15°C, cell concentration by centrifugation, solution in 50 mM Tris-HCl (pH 7.6) and 10 mM EDTA, disruption by sonication, centrifugation, SDS-PAGE	Cypridina noctiluca

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
Cypridina luciferin + O2	fusion enzyme + 10 mM luciferin (dissolved in ethanol in 30 mM Tris-HCl (pH 7.6) with 10 mM EDTA) with ATP (250 mM), CoA (250 microM), and MgCl2 (5 mM) in 100 mM Tris-HCl, pH 7.8	Cypridina noctiluca	?	Cypridina luciferase fusion enzyme expressed in Escherichia coli is soluble but does not exhibit bioluminescence	?

Synonyms

Synonyms	Comment	Organism
Cypridina luciferase	-	Cypridina noctiluca
Vargula luciferase	-	Cypridina noctiluca

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Release 2023.1 (January 2023)