A. Lehmacher et R. Hensel, CLONING, SEQUENCING AND EXPRESSION OF THE GENE ENCODING 2-PHOSPHOGLYCERATE KINASE FROM METHANOTHERMUS-FERVIDUS, MGG. Molecular & general genetics, 242(2), 1994, pp. 163-168
The gene encoding 2-phosphoglycerate kinase (2PGK), which catalyses th
e first step in the biosynthesis of cyclic 2,3-diphosphoglycerate in m
ethanogens, was cloned and sequenced from the hyperthermophilic Methan
othermus fervidus. The 2pgk gene codes for 304 amino acids, correspond
ing to a relative molecular mass of 35040. The 2pgk mRNA was estimated
to be 1600 nucleotides in size. Putative transcription signals and th
e ribosome-binding site of 2pgk are discussed. Production of 2PGK from
M. fervidus in Es-cherichia coli reveals the same apparent molecular
weights for the native enzyme and its denatured subunit as those shown
by the 2PGK purified from M. feruidus, Also the kinetic parameters of
2PKG produced in E. coli correspond well with those from the enzyme i
solated from the natural host M. fervidus.