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

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.

Keywords: carbon entry point, substrate utilization, recombinant protein, flux distribution, amino acid synthesis