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<h4>Engineering for the desired enzyme catalytic properties plays an important role in the biosynthesis of bulk chemicals and natural products. However, it is a time-consuming task to improve enzyme catalysis by traditional random mutagenesis. And the utility of rational design based on protein structure often was limited by the lack of protein structure for target enzymes and professional backgrounds of bioinformatics.<br> | <h4>Engineering for the desired enzyme catalytic properties plays an important role in the biosynthesis of bulk chemicals and natural products. However, it is a time-consuming task to improve enzyme catalysis by traditional random mutagenesis. And the utility of rational design based on protein structure often was limited by the lack of protein structure for target enzymes and professional backgrounds of bioinformatics.<br> | ||
ceaS2 enzyme is the most important enzyme in our entire acrylic acid synthesis pathway, but the activity of wild type is not high. So it is exceedingly necessary to modify it on the basis of the "part" level to improve its catalytic reactivity. We used the AEMD platform to conduct the mutational design for ceaS2 enzyme in order to figure out a more accurate scheme of mutation, which can also exert great beneficial impact on the later experiments. <br> | ceaS2 enzyme is the most important enzyme in our entire acrylic acid synthesis pathway, but the activity of wild type is not high. So it is exceedingly necessary to modify it on the basis of the "part" level to improve its catalytic reactivity. We used the AEMD platform to conduct the mutational design for ceaS2 enzyme in order to figure out a more accurate scheme of mutation, which can also exert great beneficial impact on the later experiments. <br> | ||
− | We have totally identified | + | We have totally identified 32 mutational sites, and its point mutation transformation. The experimental results show that there are XX sites, where the enzyme activity gets boosted, after the transformation. Compared to wild type ceaS2 enzyme, the highest activity has increased by 11 times, whose effect is obviously noticeable. This also demonstrates the ability of this designing platform. </h4> |
<h3 style="text-align:center">Introduction</h3> | <h3 style="text-align:center">Introduction</h3> | ||
<h4>Enzyme engineering has been extensively used to optimize biocatalysts in industrial biotechnology since most of enzymes in nature prefer to organisms adaptation but not industrial production (Alvizo, et al., 2014; Ma, et al., 2009; Savile, et al., 2010). Traditionally, optimized enzymes were obtained by random site-directed or saturated mutagenesis such as Error Prone PCR, DNA shuffling and so on (Kabumoto, et al., 2009; Qi, et al., 2009; Reetz and Carballeira, 2007; Yep, et al., 2008). Due to the immense possibility of sequence mutation at amino acids level, it is a time-consuming and low efficiency task to obtain a high efficient biocatalyst by random mutation. <br> | <h4>Enzyme engineering has been extensively used to optimize biocatalysts in industrial biotechnology since most of enzymes in nature prefer to organisms adaptation but not industrial production (Alvizo, et al., 2014; Ma, et al., 2009; Savile, et al., 2010). Traditionally, optimized enzymes were obtained by random site-directed or saturated mutagenesis such as Error Prone PCR, DNA shuffling and so on (Kabumoto, et al., 2009; Qi, et al., 2009; Reetz and Carballeira, 2007; Yep, et al., 2008). Due to the immense possibility of sequence mutation at amino acids level, it is a time-consuming and low efficiency task to obtain a high efficient biocatalyst by random mutation. <br> |
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