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Cloning, overexpression and isolation of the type II FDP aldolase from E. coli for specificity study and synthetic
I Henderson1, E Garcia-Junceda, K K Liu
1Department of Chemistry, Scripps Research Institute, La Jolla, CA 92037.
Bioorganic & Medicinal Chemistry
|August 1, 1994
Abstract:
A stable overexpression E. coli strain containing the plasmid pKEN 2 for the production of the Zn(2+)-dependent FDP aldolase from E. coli has been developed. Approximately 14,000 U of the enzyme (specific activity = 23.3 U/mg) can be obtained from 4-L of growth medium. The enzyme was isolated, purified to homogeneity and used for the studies of stability, substrate specificity and metal ion replacement and dissociation. Crystals of the enzyme have been obtained for structural analysis. This E. coli strain was deposited with the American Type Culture Collection (ATCC #77472).