The Center of Analysis and Measurement, Hunan Agricultural University, Changsha, China.
Appl Biochem Biotechnol. 2011 Jan;163(1):186-94. doi: 10.1007/s12010-010-9027-8. Epub 2010 Jul 18.
In this work, we engineered the α/β fold of mannanase Man23 based on its molecular structure analysis to obtain more stable variants. By introducing 31 single-site mutations in the α/β fold and shuffling them, the incorporation of four mutations (K178R, K207R, N340R, and S354R) displayed a good balance between high activity and stability at higher temperature and broader pH. This quartet variant was characterized by an almost threefold increased activity and a sevenfold increased stability compared to native mannanase Man23. Our results suggest that such work is safe to increase our target protein stability with no loss of activity.
在这项工作中，我们根据甘露聚糖酶 Man23 的分子结构分析对其α/β 折叠进行了工程改造，以获得更稳定的变体。通过在 α/β 折叠中引入 31 个单点突变并对其进行混合，四个突变（K178R、K207R、N340R 和 S354R）的掺入在高温和更宽的 pH 值下表现出良好的高活性和高稳定性之间的平衡。与天然甘露聚糖酶 Man23 相比，该四重变体的活性几乎增加了三倍，稳定性增加了七倍。我们的结果表明，这种工作是安全的，可以增加我们的目标蛋白稳定性，而不会损失活性。
