title: Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli.
abstract: α-Galactosidase (α-PsGal) of the cold-adapted marine bacterium Pseudoalteromonas sp. KMM 701 was cloned into the pET-40b(+) vector to study its properties and to develop an effective method for modifying human B-erythrocytes into O-blood group. The use of heat-shock as a pre-induction treatment, IPTG concentration of 0.2 mM and post-induction cultivation at 18 °C for 20 h in the developed MX-medium allowed increasing the recombinant Escherichia coli Rosetta (DE3)/40Gal strain productivity up to 30 times and the total soluble α-PsGal yield up to 40 times.
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia; Far-Eastern Federal University, Vladivostok, Russia. Electronic address: golotin@bk.ru.
authors.forename: V A
authors.initials: VA
authors.lastname: Golotin
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia; Far-Eastern Federal University, Vladivostok, Russia.
authors.forename: L A
authors.initials: LA
authors.lastname: Balabanova
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia.
authors.forename: Yu A
authors.initials: YA
authors.lastname: Noskova
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia.
authors.forename: L V
authors.initials: LV
authors.lastname: Slepchenko
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia.
authors.forename: I Yu
authors.initials: IY
authors.lastname: Bakunina
authors.affiliation: Far-Eastern Federal University, Vladivostok, Russia.
authors.forename: N S
authors.initials: NS
authors.lastname: Vorobieva
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia.
authors.forename: N A
authors.initials: NA
authors.lastname: Terenteva
authors.affiliation: G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS, Vladivostok, Russia.
authors.forename: V A
authors.initials: VA
authors.lastname: Rasskazov
mesh_terms: D000064:Acclimatization; D018929:Cell Culture Techniques / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods*; D003080:Cold Temperature; D004912:Erythrocytes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018869:Heat-Shock Response; D006801:Humans; D031161:Pseudoalteromonas / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000519:alpha-Galactosidase / Q000235:genetics* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D000519:alpha-Galactosidase
keywords: Alpha-galactosidase; Prokaryotic gene expression; Pseudoalteromonas sp.; Psychrophilic enzyme