Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli.

Authors :: Golotin VA
Balabanova LA
Noskova YA
Slepchenko LV
Bakunina IY
Vorobieva NS
Terenteva NA
Rasskazov VA
Source :: Protein expression and purification [Protein Expr Purif] 2016 Jul; Vol. 123, pp. 14-8. Date of Electronic Publication: 2016 Mar 23.
Publication Year :: 2016
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.<br /> (Copyright © 2016 Elsevier Inc. All rights reserved.)

Subjects :: Acclimatization
Cell Culture Techniques methods
Cold Temperature
Erythrocytes metabolism
Escherichia coli growth & development
Escherichia coli metabolism
Heat-Shock Response
Humans
Pseudoalteromonas genetics
Recombinant Proteins genetics
Recombinant Proteins metabolism
alpha-Galactosidase metabolism
Cloning, Molecular methods
Escherichia coli genetics
Pseudoalteromonas enzymology
alpha-Galactosidase genetics

Full Text Access

Tools