Cloning, Overexpression, Purification, and Characterization of S-Adenosylhomocysteine Hydrolase from Corynebacterium efficiens YS-314
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- 作者：J. D. Lozada-Ramí ; rez ; I. Martí ; nez-Martí ; nez ; F. Garcí ; a-Carmona ; A. Sá ; nchez-Ferrer
- 刊名：Biotechnology Progress
- 出版年：2008
- 出版时间：February 2008
- 年：2008
- 卷：24
- 期：1
- 页码：120 - 127
- 全文大小：397K
- 年卷期：v.24,no.1(February 2008)
- ISSN：1520-6033
文摘
The gene encoding S-adenosylhomocysteine hydrolase activity (SAHase: EC 3.3.1.1) fromCorynebacterium 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.
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.