Using the information available in the sequences of well-characterized transglycosidases found in plants, mutations were introduced in the glycoside hydrolase of the bacterium Thermus thermophilus, with the aim of turning it into an efficient transglycosidase. All mutants happen to have fair catalytic efficiencies, being at worst 25 times less efficient than the wild type. Noteworthy, W120F, one of our high transglycosylation yield (≈ 50%) mutants, is only two times less efficient than the wild type. Interestingly, while in the wild type the sidechain of the acid-base is only found able to sample a pair of equivalent conformations during 0.5-μs-long molecular dynamics simulations, its flexibility is much higher in the case of the high transglycosylation yield mutants. Our results thus suggest that engineering the flexibility of the acid-base of a retaining glycoside hydrolase could be a general way to turn it into an efficient transglycosidase.

中文翻译：

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致力于设计高效的糖苷酶：以嗜热栖热菌（Thermus thermophilus）GH1为例。

利用植物中发现的特征明确的转糖苷酶序列中的可用信息，将突变引入嗜热菌嗜热菌的糖苷水解酶中，目的是将其转变为有效的转糖苷酶。所有突变体碰巧都具有相当的催化效率，最差的效率是野生型的25倍。值得注意的是，W120F是我们高转糖基化产率（约50％）的突变体之一，其效率仅比野生型低两倍。有趣的是，虽然在野生型中仅发现酸碱侧链能够在0.5μs长的分子动力学模拟过程中采样一对等效构象，但在高转糖基化产率突变体的情况下，其灵活性要高得多。