An effective cold inducible expression system developed in Pseudoalteromonas haloplanktis TAC125.
A regulative two-component system previously identified in Pseudoalteromonas haloplanktis TAC125 was used to construct a cold inducible expression system that is under the control of l-malate. Performances of the inducible system were tested for both psychrophilic and mesophilic protein production using two "difficult" proteins as control. The results obtained demonstrated that both psychrophilic beta-galactosidase and yeast alpha-glucosidase are produced in a fully soluble and catalytically competent form. Optimal conditions for protein production, including growth temperature, growth medium and l-malate concentration were also investigated. Under optimized conditions yields of 620 and 27 mg/l were obtained for beta-galactosidase and alpha-glucosidase, respectively.
Papa
Rosanna
R
Department of Organic Chemistry and Biochemistry, Federico II University of Naples, Napoli, Italy.
Rippa
Valentina
V
Sannia
Giovanni
G
Marino
Gennaro
G
Duilio
Angela
A
D002384:Catalysis; D003080:Cold Temperature; D004269:DNA, Bacterial; D004790:Enzyme Induction; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D016680:Genome, Bacterial; D005959:Glucosidases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D031161:Pseudoalteromonas / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D005959:Glucosidases; D001616:beta-Galactosidase