Recording date:

Duration:

Session:

Speaker:

Co-Authors:

Abstract:

Keywords

Broad-host-range tool, inducible promoter, chassis screening

Abstract

Microbes have proven extremely valuable for the bioproduction of proteins and compounds, with applications in medical, environmental, and industrial contexts. Even though a vast number of bacterial species has been identified, the process of choosing the most suitable one as a host for a desired  application still needs optimization. A barrier arises from the limited availability of genetic engineering approaches suitable for a broad range of organisms. This hinders comparison between model organisms and restricts the consideration of non-model organisms, often overlooked due to the lack of  functional engineering tools. In this study, it is provided a modular, single vector-based expression platform, centered around the well-known promoter system tetR-pTet, inducible by anhydrotetracycline. This system has been presented in several studies with different modifications to improve its  functionality in specific organisms. However, here, we prove that one single version can be compatible with a wide range of bacteria. In all the studied microbes, the promoter system was shown to be tight and titratable, within an 84-fold dynamic range. It enables easy screening of recombinant proteins  and pathways in both mesophilic and thermophilic, and Gram-negative and Gram-positive hosts. Overall, this platform enables simple screening of heterologous expression and production in a broad variety of hosts.