Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120.
Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120.
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改善鱼腥藻 PCC 7120 脂氧合酶热稳定性的共识设计
DOI：
10.1186/s12896-018-0468-4
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发表时间：
2018-09-20
影响因子：
3.5
通讯作者：
Yang GY
中科院分区：
文献类型：
--
作者：
Qian H;Zhang C;Lu Z;Xia B;Bie X;Zhao H;Lu F;Yang GY
来自鱼腥藻PCC 7120（Ana-rLOX）的脂氧合酶（LOX）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学特性。然而，LOX的工业应用受到其差的热稳定性的限制。在此，我们报告了一个基于生物信息学的共识概念的方法，工程热稳定Ana-rLOX。一系列突变（N130 D、G260 A、S437 T、N130 D/G260 Q、N130 D/S437 Y）显示出比野生型酶更高的热稳定性和活性。因此，N130 D/G260 Q显示半衰期增加6.6倍，解折叠温度增加2.45 °C; N130 D/S437 Y显示最佳温度增加10 °C。利用圆二色性详细研究了二级结构没有太大变化，这有助于改善热稳定性。同源建模表明，增强的热稳定性和比活性可能是由于有利的疏水相互作用。通过半理性突变，在有限的结构信息下，实现了一系列突变，显示出比野生型酶更高的热稳定性和活性。我们的研究结果提供了重要的新的见解，旨在提高Ana-rLOX热稳定性和活性的分子修饰。本文的在线版本（10.1186/s12896-018-0468-4）包含补充材料，可供授权用户使用。
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. The online version of this article (10.1186/s12896-018-0468-4) contains supplementary material, which is available to authorized users.
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DOI：
10.1016/j.bbalip.2014.10.002
发表时间：
2015-04
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Kuhn H;Banthiya S;van Leyen K
通讯作者：
van Leyen K
影响因子：
14.8
作者：
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通讯作者：
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影响因子：
4.7
作者：
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通讯作者：
Bommarius, Andreas S.
影响因子：
3
作者：
Diao, Hanwen;Zhang, Chong;Lu, Zhaoxin
通讯作者：
Lu, Zhaoxin
影响因子：
3.2
作者：
Vazquez-Figueroa, Eduardo;Chaparro-Riggers, Javier;Bommarius, Andreas S.
通讯作者：
Bommarius, Andreas S.