Cobucci-Ponzano Beatrice, Perugino Giuseppe, Strazzulli Andrea, Rossi Mosè, Moracci Marco
Institute of Protein Biochemistry, National Research Council, Naples, Italy.
Methods Enzymol. 2012;510:273-300. doi: 10.1016/B978-0-12-415931-0.00015-X.
Glycosynthases are engineered glycoside hydrolases that in suitable reaction conditions promote the synthesis of oligosaccharides with exquisite stereoselectivity and enhanced regioselectivity, if compared to traditional chemical methods. This approach was demonstrated to be successful in a number of cases including β-glycosynthases acting at the termini or within an oligosaccharide chain (exo- and endo-glycosynthases, respectively) and, more recently, α-glycosynthases. This led to the production of a vast repertoire of products that include poly- and oligosaccharides, glycoconjugates, and glycopeptides. These molecules can be used as ligands of glycoside hydrolases, for the characterization of therapeutic enzymes, and as leads of drugs for the pharmaceutical industry. In this panorama, hyperthermophilic organisms, which thrive at temperatures as high as 80°C, which usually impede the growth of other living forms, have been used in the development of interesting novel glycosynthases. In fact, the extreme stability of these catalysts to extremes of pH and high concentrations of organics has allowed the exploration of novel reaction conditions, revealing new avenues for enzyme-catalyzed oligosaccharide synthesis.
糖基合成酶是经过改造的糖苷水解酶,在合适的反应条件下,与传统化学方法相比,能以优异的立体选择性和增强的区域选择性促进寡糖的合成。这种方法在许多情况下都被证明是成功的,包括作用于寡糖链末端或内部的β-糖基合成酶(分别为外切和内切糖基合成酶),以及最近的α-糖基合成酶。这导致了大量产物的产生,包括多糖和寡糖、糖缀合物和糖肽。这些分子可用作糖苷水解酶的配体,用于治疗性酶的表征,以及作为制药行业药物的先导化合物。在这种情况下,嗜热生物在高达80°C的温度下茁壮成长,而这种温度通常会阻碍其他生物的生长,已被用于开发有趣的新型糖基合成酶。事实上,这些催化剂对极端pH值和高浓度有机物具有极高的稳定性,这使得人们能够探索新的反应条件,为酶催化寡糖合成开辟了新途径。