Labeled nucleotide analogs having protein shields
US-2015330987-A1 · Nov 19, 2015 · US
US9657279B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9657279-B2 |
| Application number | US-74553908-A |
| Country | US |
| Kind code | B2 |
| Filing date | Nov 26, 2008 |
| Priority date | Nov 30, 2007 |
| Publication date | May 23, 2017 |
| Grant date | May 23, 2017 |
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The invention relates to systems and methods for production of compounds by yeast and other organisms. In one approach yeast engineered for production of a compound of commercial value is cultured together with a cellulosic bacteria, and the yeast uses a metabolic product produced by the bacteria as a carbon source. Methyl halides are an example of compounds that may be produced by this process. The invention also relates to production of organic compounds using genetically engineered organisms expressing a S-adenosylmethionine (SAM)-dependent methyl halide transferase. In one approach the organism, halides and a carbon source are incubated in a cultivation medium under conditions in which methyl halide is produced. The methyl halide may be collected and converted into non-halogenated organic molecules.
Opening claim text (preview).
The invention claimed is: 1. A co-culture system comprising a culture medium and (i) a cellulosic bacterium component, wherein the bacteria metabolize cellulose and produce one or more metabolic products, (ii) a cellulosic material or cellulose source, and (iii) a yeast component, wherein the yeast uses at least one metabolic product of the bacteria as a carbon source and produces a product different from the one or more metabolic products in (i), wherein the at least one metabolic product is selected from the group consisting of ethanol, acetate, lactate, succinate, citrate, formate, and malate, wherein growth of the bacteria is inhibited by the at least one metabolic product, wherein the bacteria and yeast have a symbiotic relationship in culture and grow together while maintaining a relatively constant ratio of species populations such that neither microorganism overtakes the other, wherein the yeast is recombinantly modified to express a heterologous protein or over-express an endogenous protein or is a recombinantly modified to knock out expression of an endogenous protein, and wherein the yeast is Saccharomyces cerevisiae and the bacteria is Actinotalea fermentans. 2. The co-culture of claim 1 that comprises cellulose and wherein the yeast is metabolically incapable of degrading cellulose. 3. The co-culture system of claim 1 in which the heterologous protein is a methyl halide transferase. 4. A yeast culture method comprising culturing cellulosic bacteria and yeast together in a liquid culture medium in the presence of cellulose or a cellulose-source, under conditions in which: (i) the bacteria metabolize cellulose and produce one or more metabolic products, and, (ii) the yeast component uses at least one metabolic product of the bacteria as a carbon source and produces a product different from the one or more metabolic products in (i) wherein the at least one metabolic product is selected from the group consisting of ethanol, acetate, lactate, succinate, citrate, formate, and malate, wherein growth of the bacteria is inhibited by the at least one metabolic product, wherein the bacteria and yeast have a symbiotic relationship in culture and grow together while maintaining a relatively constant ratio of species populations, and wherein the yeast is S. cerevisiae and the bacteria is Actinotalea fermentans. 5. The method of claim 4 wherein the yeast is recombinantly modified to express a heterologous protein. 6. The method of claim 5 in which the heterologous protein is a methyl halide transferase. 7. The method of claim 4 further comprising recovering a product from the culture medium which product is produced by the yeast. 8. The method of claim 7 wherein the product is a methyl halide. 9. A method for production of methyhalide comprising culturing a cellulosic bacteria which metabolizes cellulose and produces one or more metabolic products together with a yeast which does not metabolize cellulose and which is recombinantly modified to express a heterologous methyl halide transferase protein in a medium containing a cellulose source and a halide, under conditions in which methyl halide is produced wherein the yeast is S. cerevisiae and the bacterium is Actinotalea fermentans. 10. The method of claim 9 wherein the carbon source is ethanol, acetate, lactate, succinate, formate, citrate, or malate. 11. The method of claim 9 further comprising recovering methyl halide from the culture medium. 12. The method of claim 11 further comprising the step of converting the methyl halide into a non-halogenated organic molecule or a mixture of non-halogenated organic molecules. 13. The co-culture system of claim 1 , wherein the co-culture system comprises one species of yeast and one species of bacteria.
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