Abstract
Efficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is the fastest growing free-living bacterium with a doubling time of less than 10 min, which makes it highly attractive as a protein expression host. Here, 196 pET plasmids with different genes of interest (GOIs) were electroporated into the V. natriegens strain VnDX, which carries an integrated T7 RNA polymerase expression cassette. As a result, 65 and 75% of the tested GOIs obtained soluble expression in V. natriegens and Escherichia coli, respectively, 20 GOIs of which showed better expression in the former. Furthermore, we have adapted a consensus “what to try first” protocol for V. natriegens based on Terrific Broth medium. Six sampled GOIs encoding biocatalysts enzymes thus achieved 50–128% higher catalytic efficiency under the optimized expression conditions. Our study demonstrated V. natriegens as a pET-compatible expression host with a spectrum of highly expressed GOIs distinct from E. coli and an easy-to-use consensus protocol, solving the problem that some GOIs cannot be expressed well in E. coli.
Highlights
The Escherichia coli pET expression system remains one of the most popular systems for recombinant protein production (Shilling et al, 2020)
This study compared the soluble expression of 196 genes of interest (GOIs) encoding enzymes of various families from various sources between V. natriegens VnDX and E. coli BL21(DE3)
The results showed that the engineered V. natriegens strain VnDX is compatible with the pET system but had a distinct spectrum of highly expressed proteins compared with E. coli, indicating that it might be a valuable complementary expression system to E. coli
Summary
The Escherichia coli pET expression system remains one of the most popular systems for recombinant protein production (Shilling et al, 2020). Numerous strategies have been proposed to improve recombinant protein production, including codon optimization of target proteins (Burgess-Brown et al, 2008; Menzella, 2011; Chung and Lee, 2012), screening and construction of a superior host cell (Choi et al, 2015; Liu et al, 2015; Hayat et al, 2018), addition of fusion tags (Esposito and Chatterjee, 2006; Young et al, 2012; Hayat et al, 2018), optimization of expression elements (Gustafsson et al, 2012; Eichmann et al, 2019), or the development of high-density fermentation processes (Yee and Blanch, 1992; Zhang et al, 2000; Thongekkaew et al, 2008). V. natriegens as the host for GOI expression is allowed to own higher productivity, saving time and energy costs in industrial production (Hoffart et al, 2017)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
