Department of Chemical and Biological Sciences, School of Sciences, Universidad de las Américas Puebla, Santa Catarina Mártir Cholula 72820, Puebla, México.
Appl Biochem Biotechnol. 2013 Jun;170(3):639-53. doi: 10.1007/s12010-013-0218-y. Epub 2013 Apr 16.
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-腺苷同型半胱氨酸水解酶（SAHase）由 sahase 基因编码，是大多数生物中催化腺苷和同型半胱氨酸可逆转化为 S-腺苷同型半胱氨酸的关键酶。sahase 基因是从高度耐热的厌氧细菌 Thermotoga maritima 的基因组中分离出来的，然后通过大肠杆菌进行克隆、鉴定、过表达，并通过热沉淀进行部分纯化。重组 SAHase 的热纯化导致圆二色性光谱发生变化。通过该分析，可以确定纯化后重组酶的α-螺旋和β-折叠含量组成的结构变化。此外，通过比较分子建模进行了预测的二级结构和 3D 结构模型，以进一步了解该蛋白质的分子功能，包括其有吸引力的生物技术用途。
