Examining How Different Carbon Entry Point Affects Recombinant Protein Production from Ethylene Glycol in Bacillus Subtilis

Wenfa Ng
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Abstract

Recombinant protein production is the first application task for nascent genetic engineering efforts in 1970s. Since then, a variety of basic science and process engineering approaches have been used in improving recombinant protein production. Given that protein production could serve as marker for how well rewired metabolism is functioning, possibility exists in using it to highlight the most suitable pathway for assimilating a particular unconventional substrate. To this end, different pathways with different carbon entry point in central carbon metabolism could be constructed, with fluorescence of a green fluorescent protein as readout. Good carbon entry point should ideally lead to even distribution of substrate flux to all precursors feeding amino acid synthesis, and should result in strong fluorescence intensity and high protein production. This perspective critically examines the literature for prior work on the above hypothesis in relation to utilization of ethylene glycol by Bacillus subtilis. A literature review suggests that the hypothesis is novel, and provides additional information that modulation of global regulatory protein (transcription factor) may serve as ancillary factors promoting high recombinant protein production.
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探讨枯草芽孢杆菌中不同碳入口点对乙二醇生产重组蛋白的影响
重组蛋白的生产是20世纪70年代新生的基因工程努力的第一个应用任务。从那时起,各种基础科学和工艺工程方法被用于改善重组蛋白的生产。考虑到蛋白质的产生可以作为重新连接代谢功能的标志,利用它来突出吸收特定非常规底物的最合适途径是存在可能性的。为此,可以构建中心碳代谢不同碳入口点的不同途径,以绿色荧光蛋白的荧光为读出。理想情况下,良好的碳入口点应导致底物通量均匀分布到所有用于氨基酸合成的前体,并应导致强荧光强度和高蛋白质产量。这一观点批判性地检查了文献先前的工作对上述假设有关利用乙二醇的枯草芽孢杆菌。文献综述表明,这一假设是新颖的,并提供了额外的信息,即全局调节蛋白(转录因子)的调节可能是促进高重组蛋白生产的辅助因素。
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