Applied Biochemistry and Biotechnology

, Volume 170, Issue 3, pp 639–653 | Cite as

Recombinant S-Adenosylhomocysteine Hydrolase from Thermotoga maritima: Cloning, Overexpression, Characterization, and Thermal Purification Studies

  • J. D. Lozada-RamírezEmail author
  • A. Sánchez-Ferrer
  • F. García-Carmona


S-Adenosylhomocysteine hydrolase (SAHase) encoded by sahase gene is a determinant when catalyzing the reversible conversion of adenosine and homocysteine to S-adenosylhomocysteine in most living organisms. The sahase gene was isolated from the genome of the highly thermostable anaerobic bacteria Thermotoga maritima, and then it was cloned, characterized, overexpressed using Escherichia coli, and partially purified by thermal precipitation. The thermal purification of the recombinant SAHase resulted in changes in the circular dichroism spectra. As a result of this analysis, it was possible to determine the structural changes in the composition of the α-helix and β-sheet content of the recombinant enzyme after purification. Moreover, a predicted secondary structure and 3D structural model was rendered by comparative molecular modeling to further understand the molecular function of this protein including its attractive biotechnological use.


S-Adenosylhomocysteine hydrolase S-Adenosyl-l-homocysteine Thermal purification Thermotoga maritima Structural modeling 



S-Adenosyl-l-homocysteine hydrolase


Recombinant S-adenosyl-l-homocysteine hydrolase










Circular dichroism




S-Adenosylhomocysteine hydrolase coding gene



This study was partially supported by Spanish grants from MINECO-FEDER (BIO2010-22225-C02-01) and from Programa de Ayuda a Grupos de Excelencia de la Región de Murcia, Fundación Séneca (04541/GERM/06, Plan Regional de Ciencia y Tecnología 2007–2010), and by Proyecto CONACYT Ciencia Básica 2009–2012 (CB-133949).

Supplementary material

12010_2013_218_MOESM1_ESM.jpg (104 kb)
Additional material 1 (A) 3D structural model of a single unit of SAHase from Thermotoga maritima. Alpha-helix are indicated in black, beta-strand in gray, and random coils in white. (B) Amino acid residues of the conserved domains implied in SAHase catalytic activity (Ado binding), and in NAD+ binding. Amino acid residues are discussed in the text. (JPEG 103 kb)
12010_2013_218_MOESM2_ESM.jpg (175 kb)
Additional material 2 Ribbon diagrams of the tetrameric structure of rat SAHase (A) and the theoretical structure of T. maritima SAHase. (B) Enlargements. (C) and (D) Amino acid residues which are involved in the network interactions in the central channel of rat SAHase and T. maritima SAHase, respectively. Amino acid residues are discussed in the text. (JPEG 175 kb)


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • J. D. Lozada-Ramírez
    • 1
    Email author
  • A. Sánchez-Ferrer
    • 2
  • F. García-Carmona
    • 2
  1. 1.Department of Chemical and Biological Sciences, School of SciencesUniversidad de las Américas PueblaSanta Catarina Mártir CholulaMéxico
  2. 2.Department of Biochemistry and Molecular Biology-A, Faculty of BiologyUniversity of MurciaMurciaSpain

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