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Synthesizing a Cellulase like Chimeric Protein by Recombinant Molecular Biology Techniques

机译:通过重组分子生物学技术合成纤维素酶像嵌合蛋白

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摘要

In order to meet the Renewable Fuels Standard demands for 30 billion gallons of biofuels by the end of 2020, new technologies for generation of cellulosic ethanol must be exploited. Breaking down cellulose by cellulase enzyme is very important for this purpose but this is not thermostable and degrades at higher temperatures in bioreactors. Towards creation of a more ecologically friendly method of rendering bioethanol from cellulosic waste, we attempted to produce recombinant higher temperature resistant cellulases for use in bioreactors. The project involved molecular cloning of genes for cellulose-degrading enzymes based on bacterial source, expressing the recombinant proteins in E. coli and optimizing enzymatic activity. We were able to generate in vitro bacterial expression systems to produce recombinant His-tag purified protein which showed cellulase like activity.
机译:为了满足到2020年底可再生燃料标准对300亿加仑生物燃料的需求,必须开发用于生产纤维素乙醇的新技术。为此目的,通过纤维素酶分解纤维素非常重要,但这不是热稳定的,并且在生物反应器中会在较高温度下降解。为了创造一种从纤维素废料中提取生物乙醇的更加生态友好的方法,我们试图生产用于生物反应器的重组耐高温纤维素酶。该项目涉及基于细菌来源的纤维素降解酶基因的分子克隆,在大肠杆菌中表达重组蛋白并优化酶活性。我们能够产生体外细菌表达系统,以产生重组的His标签纯化的蛋白,该蛋白显示出纤维素酶样活性。

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