Structure-guided consensus approach to create a more thermostable penicillin G acylase
Structure-guided consensus approach to create a more thermostable penicillin G acylase
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DOI：
10.1002/biot.200600029
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发表时间：
2006-05-01
影响因子：
4.7
通讯作者：
Bommarius, Andreas S.
中科院分区：
文献类型：
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作者：
Polizzi, Karen M.;Chaparro-Riggers, Javier F.;Bommarius, Andreas S.
青霉素G酰化酶（PGA）由于其体积大、成熟过程复杂，其热稳定化一直是一个难题。我们开发了一种数据驱动的蛋白质设计方法，该方法比传统的共识方法需要更少的同源序列，并利用结构信息来限制创建的变体数量。实验发现，在21个单点突变体中，约有50%的突变体比野生型PGA更耐热，其中两个突变体在50摄氏度时的半衰期几乎是野生型PGA的三倍，对活性几乎没有影响。对四个预测点突变热稳定性的程序的分析表明，程序与实验数据之间的一致性很小，强调所选择的稳定突变很难预测，但我们的数据驱动设计方法应该证明是有用的。
The thermostabilization of penicillin G acylase (PGA) is a difficult problem due to the large size of the protein and its complex maturation process. We developed a data-driven protein design method that requires fewer homologous sequences than the traditional consensus approach and utilizes structural information to limit the number of variants created. Approximately 50% of our 21 single-point mutants were found experimentally to be more thermostable than the wild-type PGA, two had almost threefold longer half-life at 50 degrees C, with very little effect on activity. An analysis of four programs that predict the thermostability conferred by point mutations shows little agreement between the programs and with the experimental data, emphasizing that the chosen stabilizing mutations are very difficult to predict, but that our data-driven design method should prove useful.