Abstract

Membrane proteins represent a class of proteins that are difficult targets to characterize. Their structural and functional characterization requires that they first be produced at quantities that enable their biophysical and biochemical analysis. Because they are natively produced at levels much lower than their soluble counterparts, extraction from their natural sources is not sufficient to produce enough material for these studies. Recombinant protein expression and production has become a popular method to produce large amounts of proteins for research and industrial purposes. Significant effort has been spent finding new ways to optimize and increase protein expression. As cutting edge techniques in synthetic biology continue to advance they offer a potential well of opportunities to tune expression through better control of the transcription and translation processes. Many techniques being developed are geared toward the production of soluble proteins, but in the following review, a focus on effective strategies to maximize membrane protein production in yeast is presented and includes many of the most innovative approaches to maximize expression using synthetic biology. Synthetic biology utilizes modern techniques in molecular biology and genetic engineering to optimize the production of compounds produced in microbes by altering gene elements required for transcription and translation of critical genes responsible for their synthesis. Compounds include natural products, hydrocarbon-based compounds for biofuels, and therapeutic proteins. Producing membrane proteins recombinantly using similar methods to increase expression yields is described in this review along with cutting edge techniques like cell-free expression, which circumvents many of the common problems that plague overexpression of membrane proteins microbial-based platforms.