Tailor‐made biocatalysts enzymes for the fine chemical industry in China

Abstract

The Center of Industrial Biotechnology (CIBT) was established in Huzhou for fine chemicals in 2006 and CIBT Shanghai was founded for bulk chemicals in 2008. CIBT is a non‐profit organization under auspices of the Shanghai Institutes for Biological Sciences, Shanghai Branch of the Chinese Academy of Sciences (CAS) and Huzhou Municipal Government. CIBT is affiliated with the CAS, which enables it to take advantage of the rich R&D resources and support from CAS; yet CIBT operates as an independent legal entity. The goal of CIBT is to incubate industrial biotechnologies and accelerate the commercialization of these technologies with corporate partners in China.

Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli

“…Because different proteins had different optimal expression conditions, there can be no "right" answer a priori (Gräslund et al, 2008), which is the same in V. natriegens (Kormanova et al, 2020). In order to facilitate the decision-making, we compared with the general fermentation method for E. coli established in our laboratory (Jiang et al, 2016), proposed a consensus "what to try first" protocol for V. natriegens and verified the effectiveness of this protocol in the expression of multiple GOIs. This protocol provides a reference for researchers who are starting out with protein expression in V. natriegens.…”

“…In a previous study, we established an enzyme library (Supplementary Table 1) containing 196 GOIs, including many commonly used biocatalyst enzymes (Jiang et al, 2016), the Saccharomyces cerevisiae sodium-potassium pump Trk1 (Ko and Gaber, 1991), and plant-derived isoprenoid synthase and its mutants (Yang et al, 2016), covering seven families of enzymes from bacteria, fungi, and plants. The pET expression plasmids encoding the GOIs were electroporated into V. natriegens VnDX and E. coli BL21(DE3) to compare the two expression systems.…”

“…Therefore, a more general protocol for V. natriegens was needed, giving researchers the first-to-try choice for protein production in this novel host. We aimed to adapt a consensus "what to try first" protocol in TBv2 for V. natriegens based on our collective analysis of the expression of over 200 GOIs in E. coli BL21(DE3) (Gräslund et al, 2008;Jiang et al, 2016;Rongsheng et al, 2016).…”

“…Here, we electroporated each of 196 recombinant pET plasmids expressing commonly used biocatalysts enzymes (Jiang et al, 2016) into the non-pathogenic and fastest-growing V. natriegens ATCC14048 with an integrated T7 RNA polymerase cassette in advance. Analysis of the soluble expression of these GOIs in both V. natriegens VnDX and E. coli BL21(DE3) showed that the spectrum of highly expressed GOIs in V. natriegens was complementary to that of E. coli.…”