首页> 外文会议>Conference on biochemical and molecular engineering >IMPROVING THE CATALYTIC ACTIVITY OF ISOPENTENYL PHOSPHATE KINASE THROUGH PROTEIN COEVOLUTION ANALYSIS
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IMPROVING THE CATALYTIC ACTIVITY OF ISOPENTENYL PHOSPHATE KINASE THROUGH PROTEIN COEVOLUTION ANALYSIS

机译:通过蛋白质共进化分析提高磷酸异戊酯磷酸激酶的催化活性

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Protein rational design has become more and more popular for protein engineering with the advantage of biological big-data. In this study, we described a method of rational design that is able to identify desired mutants by analyzing the coevolution of protein sequence. We employed this approach to evolve an archaeal isopentenyl phosphate kinase that can convert dimethylallyl alcohol (DMA) into precursor of isoprenoids. By designing 9 point mutations, we improved the catalytic activities of IPK about 8-fold in vitro. After introducing the optimal mutant of IPK into engineered E. coli strain for p-carotenoids production, we found that (3-carotenoids production exhibited 97% increase over the starting strain. The process of enzyme optimization presented here could be used to improve the catalytic activities of other enzymes.
机译:蛋白质理性设计与生物大数据的优势变得越来越流行的蛋白质工程。在该研究中,我们描述了一种理性设计方法,其能够通过分析蛋白质序列的共同识别所需的突变体。我们采用这种方法来发展脑面戊烯基磷酸激酶,其可以将二甲基丙醇醇(DMA)转化为异戊二烯的前体。通过设计9点突变,我们改善了IPK的催化活性约8倍。在将IPK的最佳突变体引入工程化的大肠杆菌菌株后,我们发现(3类胡萝卜素生产在起始菌株上显示出97%。此处呈现的酶优化方法可用于改善催化剂其他酶的活动。

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