Strategies for Altering Enzyme Reaction Specificity for Applied Biocatalysis
Strategies for Altering Enzyme Reaction Specificity for Applied Biocatalysis
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DOI：
10.2174/1385272053174967
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发表时间：
2005-02
影响因子：
2.6
通讯作者：
Per Berglund;Seongsoon Park
中科院分区：
文献类型：
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作者：
Per Berglund;Seongsoon Park
事实上，许多酶具有广泛的底物特异性，这是酶在合成化学中广泛应用的重要基础。此外，许多酶可以催化与天然酶完全不同的反应。利用分子生物学技术控制酶的这种催化可塑性，从而在活性位点建立全新的反应特异性的可能性是本综述的主题。这些例子被细分为六种不同的方法（i - vi）用于工程的反应特异性。第一种方法(i)是定向进化的随机方法，以获得新的反应特异性。其他方法包括将已知酶的反应特异性引入到另一个密切相关的酶中，通过替换两个酶的（ii）序列或（iii）结构重叠选择的关键氨基酸残基。然而，其他方法包括替代关键氨基酸残基以引入新的反应特异性，而无需与模板酶进行比较（iv）和引入完整的催化机制(v)。最后一种方法是在非催化蛋白中引入活性位点（vi）。这六种不同的方法改变酶的反应化学性质，每一种都是控制酶的催化可塑性的有力工具。尽管实际应用的例子很少，而且仍然具有挑战性，但这些酶作为合成化学催化剂的前景非常广阔。在改变酶的反应特异性方面取得的进展将导致朝着为合成化学创造量身定制的酶的目标不断发展。
The fact that many enzymes have broad substrate specificity has been a property, of fundamental importance for the widespread applications of enzymes in synthetic chemistry. Many enzymes can, in addition, catalyze completely different reactions compared to their natural ones. The possibility of using molecular biology techniques to control Such catalytic plasticity of enzymes in order to establish completely new reaction specificity in the active site is the topic for this review. The examples are subdivided according to six different approaches used (i - vi) for engineering of the reaction specificity. The first approach (i) is the random method of directed evolution to achieve new reaction specificity. Other approaches involve strategies where tire reaction specificity of a known enzyme is implemented into another, closely related, enzyme by substituting key amino acid residues selected either by (ii) sequence or (iii) structural overlap of the two enzymes. Yet other approaches involve substitution of key amino acid residues to introduce new reaction specificity without comparing with a template enzyme (iv) and the introduction of a complete catalytic machinery (v). The final approach is the introduction of an active site into a non-catalytic protein (vi). These six different approaches for altering the reaction chemistry of enzymes each represent a powerful tool for controlling the catalytic plasticity or enzymes. The prospect for these altered enzymes as catalysts in synthetic chemistry is very large although examples of practical use are rare and still challenging. The progress in the area of altering enzyme reaction specificity will result in a Continued development towards the goal of creating tailor-made enzymes for synthetic chemistry.