APPLICATIONS OF TECHNOLOGY:
- Improved production of C2-C6 linear alcohols and aldehydes
- Alcohol-based induction system for gene regulation and protein overexpression
ADVANTAGES:
- High throughput screening and selection
- Improved sensitivity for measuring enzyme activity and small molecule production
- Broad applicability
- Single cell resolution capabilities—individual biofuels producing microbes can be screened by fluorescence-activated cell sorting (FACS)
ABSTRACT:
Researchers at the Joint BioEnergy Institute (JBEI) have invented a method of using transcription factors expressed in vivo to evolve, screen, and select for microorganisms producing an intracellular small molecule of interest, such as a short chain alcohol. In the JBEI invention, biosensors composed of transcription factors and their cognate promoters are designed and constructed to be capable of binding the particular molecule of interest.
For the biofuels industry, an in vivo biosensor in a microorganism, such as E. coli, for the detection of short-chain alcohols can be used to develop high-throughput selections and screens to engineer microorganisms producing alcohol-based biofuel molecules. Currently, low throughput chromatography methods are used for metabolite detection. The JBEI invention improves screening throughput over current methods by several orders of magnitude. In addition, the invention exhibits greater sensitivity as compared to high-throughput colorimetric screens. Downstream application of the invention as a selection method could allow for direct, dynamic evolution of strains without the need for screening.
The JBEI technology focuses on bacterial transcription factors and their cognate promoters. This class of transcription factors has been shown to bind to a broad range of industrial and commercial molecules.
The Joint BioEnergy Institute (JBEI, www.jbei.org) is a scientific partnership led by the Lawrence Berkeley National Laboratory and including the Sandia National Laboratories, the University of California campuses of Berkeley and Davis, the Carnegie Institution for Science and the Lawrence Livermore National Laboratory. JBEI’s primary scientific mission is to advance the development of the next generation of biofuels.
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STATUS:
Issued US Patent 8552169 available at www.uspto.gov. Available for licensing or collaborative research.
REFERENCE NUMBER: EJIB-2710