The gene encoding
S-adenosylhomocysteine hydrolase activity (SAHase: EC 3.3.1.1) from
Corynebacterium efficiens (YS-314) was cloned and expressed as a fusion protein in
Escherichiacoli Rosetta (DE3). The analyzed nucleotide sequence of the cloned gene proved to be identicalto those reported on the NCBI database. The recombinant enzyme is a tetramer, showing amolecular weight of approximately 210 kDa, as estimated by gel filtration. The
KM values ofthe enzyme for
S-adenosylhomocysteine (SAH), adenosine (Ado), and homocysteine (Hcy), weredetermined to be 1.4, 10, and 45
M. The overexpression of the recombinant enzyme produceda high level of protein (>40 mg of protein per gram of wet cells) and revealed certainthermostability when characterized at temperatures above 40
C. It also showed a high capacityfor the synthesis of SAH, thermal stability, and high kinetic similarity to human SAHase,indicating a high biotechnological and pharmacological potential.