PDF(Sci-hub)
Abstract：
Lipoxygenase (LOX) from Anabaena sp. PCC 7120 (Ana-rLOX) offers important applications in the food industry, especially for improving aroma and dough rheological properties. However, industrial applications of LOXs have been limited by their poor thermostability. Herein, we report a bioinformatics-based consensus concept approach for the engineering of thermostable Ana-rLOX.
A series of mutations (N130D, G260A, S437T, N130D/G260Q, N130D/S437Y) showed higher thermostability and activity than the wild-type enzyme. Thus, N130D/G260Q exhibited a 6.6-fold increase in half-life and 2.45 °C increase in unfolding temperature; N130D/S437Y showed a 10 °C increase in optimal temperature. The secondary structure did not change much that contributed to improved thermostability were investigated in detail using circular dichroism. Homology modeling suggested that enhanced thermostability and specific activity may result from favorable hydrophobic interactions.
A series of mutations were achieved, showing higher thermostability and activity than the wild-type enzyme by semi-rational mutagenesis with limited structure information. Our findings provide important new insights into molecular modifications aimed at improving Ana-rLOX thermostability and activity.
A series of mutations (N130D, G260A, S437T, N130D/G260Q, N130D/S437Y) showed higher thermostability and activity than the wild-type enzyme. Thus, N130D/G260Q exhibited a 6.6-fold increase in half-life and 2.45 °C increase in unfolding temperature; N130D/S437Y showed a 10 °C increase in optimal temperature. The secondary structure did not change much that contributed to improved thermostability were investigated in detail using circular dichroism. Homology modeling suggested that enhanced thermostability and specific activity may result from favorable hydrophobic interactions.
A series of mutations were achieved, showing higher thermostability and activity than the wild-type enzyme by semi-rational mutagenesis with limited structure information. Our findings provide important new insights into molecular modifications aimed at improving Ana-rLOX thermostability and activity.
摘要：
脂氧合酶（LOX）来自鱼腥草属。PCC7120（Ana-rLOX）在食品工业中提供了重要的应用，特别是用于改善香气和面团流变特性。然而,LOX的工业应用受到其差的热稳定性的限制。在这里，我们报告了一种基于生物信息学的共识概念方法，用于热稳定的Ana-rLOX工程。
一系列突变（N130D，G260A,S437T,N130D/G260Q,N130D/S437Y）显示出比野生型酶更高的热稳定性和活性。因此，N130D/G260Q的半衰期增加了6.6倍，展开温度增加了2.45°C；N130D/S437Y的最佳温度增加了10°C。二级结构变化不大，这有助于提高热稳定性，使用圆二色性进行了详细研究。同源性建模表明，增强的热稳定性和比活性可能是有利的疏水相互作用的结果。
实现了一系列突变，通过具有有限结构信息的半理性诱变显示出比野生型酶更高的热稳定性和活性。我们的发现为旨在改善Ana-rLOX热稳定性和活性的分子修饰提供了重要的新见解。
一系列突变（N130D，G260A,S437T,N130D/G260Q,N130D/S437Y）显示出比野生型酶更高的热稳定性和活性。因此，N130D/G260Q的半衰期增加了6.6倍，展开温度增加了2.45°C；N130D/S437Y的最佳温度增加了10°C。二级结构变化不大，这有助于提高热稳定性，使用圆二色性进行了详细研究。同源性建模表明，增强的热稳定性和比活性可能是有利的疏水相互作用的结果。
实现了一系列突变，通过具有有限结构信息的半理性诱变显示出比野生型酶更高的热稳定性和活性。我们的发现为旨在改善Ana-rLOX热稳定性和活性的分子修饰提供了重要的新见解。
