This invention provides the genetic engineering technologies and the defined media to convert gram-positive microorganisms for a low-cost, consolidated bio-processing (CBP) platform. This technology can be applied for producing advanced biofuels and other bio-based products with lignocellulosic material as feedstock in the defined media without any expensive organic nutrients and cellulases.
The present invention provides a sonoporation-based method that can be universally applied for delivery of compounds into Gram positive bacteria. Gram positive bacteria which can be transformed by sonoporation include, for example, Bacillus, Streptococcus, Acetobacterium, and Clostridium. Compounds which can be delivered into Gram positive bacteria via sonoporation include nucleic acids (DNA or RNA), proteins, lipids, carbohydrates, viruses, small organic and inorganic molecules, and nano-particles.
Dr. Adam’s group at The University of Georgia has discovered the identity of a group of genes that enable a microorganism to convert untreated woody plant biomass, such as poplar wood chips, to soluble materials that can be used by the same organism or by another to produce biofuels, such as hydrogen and ethanol. The discovery was made from an analysis of the genomes of two very closely related microorganisms Anaerocellum thermophilum and Caldicellulosiruptor saccharolyticus, only one of which, Anaerocellum thermophilum, is able to grow on unprocessed woody plant biomass. The genes that confer this property to Anaerocellum thermophilum are termed PBU for plant biomass utilization. Many of the PBU genes are present in Anaerocellum thermophilum as gene clusters.
The present invention provides isolated nucleic acid molecules which encode a mutant acetaldehyde-CoA/alcohol dehydrogenase or mutant alcohol dehydrogenase and confer enhanced tolerance to ethanol. The invention also provides related expression vectors, genetically engineered microorganisms having enhanced tolerance to ethanol, as well as methods of making and using such genetically modified microorganisms for production of biofuels based on fermentation of biomass materials
ORNL inventors discovered a novel microorganism that displays exceptional tolerance to several growth inhibitors generated during dilute acid pretreatment of lignocellulosic biomass. Vanillin, furfural, hydroxymethylfurfural and lignin are known to inhibit the growth of microorganisms, such as yeast, during fermentation of sugars to alcohols. The new, isolated organism can be used as a source of resistance factors which could then be engineered into advanced biocatalysts for consolidated bioprocessing of biomass into fuels.
University of Georgia researchers have invented a method to more efficiently decompose biomass, which lowers the cost of producing biofuel. The method centers around a bacterium called Caldicellulosiruptor, which as has unique properties that make it more conducive for processing a type of biomass known as lignocellulsic. The modification of the bacterium DNA will improve the efficiency of converting biomass into fuels. Furthermore, the researchers have developed general procedures that can be utilized across other sections of the bacterium species.
National Renewable Energy Laboratory researchers discovery that the thermostable enzyme for C. bescii is a superior degrader of crystalline biomass compared to a traditional cellulase mixture (Cel7)/E1(Cel5) at equivalent enzyme loading. They have shown that CelA purified from C. bescii achieves a much higher extent of conversion on highly crystalline model substrates (Avicel) than traditionally used enzyme.
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