{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["van Rossum T"],"funding":["7th framework program for research and technological development (FP7) of the European Union","Graduate School VLAG"],"pagination":["625-641"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC5404197"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["10(3)"],"pubmed_abstract":["The use of bioreporters in high-throughput screening for small molecules is generally laborious and/or expensive. The technology can be simplified by coupling the generation of a desired compound to cell survival, causing only positive cells to stay in the pool of generated variants. Here, a dual selection/screening system was developed for the in vivo detection of novel biocatalysts. The sensor part of the system is based on the transcriptional regulator AraC, which controls expression of both a selection reporter (LeuB or KmR; enabling growth) for rapid reduction of the initially large library size and a screening reporter (LuxCDABE; causing bioluminescence) for further quantification of the positive variants. Of four developed systems, the best system was the medium copy system with KmR as selection reporter. As a proof of principle, the system was tested for the selection of cells expressing an l-arabinose isomerase derived from mesophilic Escherichia coli or thermophilic Geobacillus thermodenitrificans. A more than a millionfold enrichment of cells with l-arabinose isomerase activity was demonstrated by selection and exclusion of false positives by screening. This dual selection/screening system is an important step towards an improved detection method for small molecules, and thereby for finding novel biocatalysts."],"journal":["Microbial biotechnology"],"pubmed_title":["A growth- and bioluminescence-based bioreporter for the in vivo detection of novel biocatalysts."],"pmcid":["PMC5404197"],"funding_grant_id":["265933"],"pubmed_authors":["van Rossum T","Muras A","Creutzburg SCA","Kengen SWM","van der Oost J","Baur MJJ"],"additional_accession":[]},"is_claimable":false,"name":"A growth- and bioluminescence-based bioreporter for the in vivo detection of novel biocatalysts.","description":"The use of bioreporters in high-throughput screening for small molecules is generally laborious and/or expensive. The technology can be simplified by coupling the generation of a desired compound to cell survival, causing only positive cells to stay in the pool of generated variants. Here, a dual selection/screening system was developed for the in vivo detection of novel biocatalysts. The sensor part of the system is based on the transcriptional regulator AraC, which controls expression of both a selection reporter (LeuB or KmR; enabling growth) for rapid reduction of the initially large library size and a screening reporter (LuxCDABE; causing bioluminescence) for further quantification of the positive variants. Of four developed systems, the best system was the medium copy system with KmR as selection reporter. As a proof of principle, the system was tested for the selection of cells expressing an l-arabinose isomerase derived from mesophilic Escherichia coli or thermophilic Geobacillus thermodenitrificans. A more than a millionfold enrichment of cells with l-arabinose isomerase activity was demonstrated by selection and exclusion of false positives by screening. This dual selection/screening system is an important step towards an improved detection method for small molecules, and thereby for finding novel biocatalysts.","dates":{"release":"2017-01-01T00:00:00Z","publication":"2017 May","modification":"2024-02-16T12:47:53.452Z","creation":"2019-03-27T02:42:00Z"},"accession":"S-EPMC5404197","cross_references":{"pubmed":["28393499"],"doi":["10.1111/1751-7915.12612"]}}