Protein Expression and Purification, Vol.123, 14-18, 2016
Optimization of cold-adapted alpha-galactosidase expression in Escherichia coli
alpha-Galactosidase (alpha-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 degrees 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 alpha-PsGal yield up to 40 times. (C) 2016 Elsevier Inc. All rights reserved.