Cell-based systems for production of methyl formate
View Patent ↗Disclosed is a process in which a recombinant organism, such as a yeast, expressing a heterologous S-adenosylmethionine (SAM)-dependent methyl halide transferase (MHT) protein is combined with a halide and a carbon source in a cultivation medium under conditions in which methyl formate is produced. The cell may genetically modified to express methyl formate synthase, methanol dehydrogenase and/or hydrolytic dehalogenase at levels higher than a cell of the same species that is not genetically modified. The methyl formate may be collected and used in a variety of applications. The halide may be chlorine, bromine or iodine.
1. A method of producing methyl formate comprising
a) combining
i) a recombinant yeast comprising a heterologous gene encoding S-adenosylmethionine (SAM)-dependent methyl halide transferase (MHT),
ii) a halide selected from the group consisting of chlorine, bromine and iodine; and
iii) a carbon source;
in a cultivation medium under conditions in which methyl formate is produced; and
b) collecting methyl formate.
2. The method of claim 1 in which methyl halide is not collected, or the amount of methyl formate collected is at least 10-fold greater than the amount of methyl halide collected.
3. The method of claim 2 further comprising removing methyl halide from the methyl formate that is collected.
4. The method of claim 1 further comprising the step of converting the collected methyl formate into methanol.
5. The method of claim 1 wherein the organism is a yeast that is from a genus selected from the group consisting of Saccharomyces, Pichia, Hansenula, Kluyveromyces, Yarrowia, Trichoderma and Scizosacchromyces.
6. The method of claim 1 wherein the organism is a yeast that is not a methylotrophic yeast.
7. The method of claim 1 wherein MHT is from Batis maritima.
8. The method of claim 1 wherein the organism is a yeast genetically modified to increase flux through a S-adenosyl-methionine (SAM) biosynthetic pathway.
9. The method of claim 1 wherein the organism is a yeast genetically modified to express a heterologous or modified methanol dehydrogenase.
10. The method of claim 1 wherein the organism is a yeast genetically modified to express a heterologous or modified methyl formate synthase.
11. The method of claim 1 wherein the organism is a yeast genetically modified to express a heterologous or modified hydrolytic dehalogenase.
12. The method of claim 11 wherein the yeast is genetically modified to express Sphingomonas paucimobilis LinB protein.
13. The method of claim 9 wherein the organism is a yeast genetically modified to express or over-express S. cerevisia ADH4 protein and/or S. cerevisia ADH3 protein.
14. The method of claim 13 wherein the yeast is S. cerevisia modified to over-express S. cerevisia ADH4 protein and/or S. cerevisia ADH3 protein.
15. A genetically engineered yeast cell expressing a heterologous S adenosylmethionine (SAM)-dependent methyl halide transferase (MHT) and expressing one, two or three of:
a) a heterologous methyl formate synthase,
b) a heterologous methanol dehydrogenase,
c) a heterologous hydrolytic dehalogenase.
16. A genetically engineered yeast cell expressing a heterologous S adenosylmethionine (SAM)-dependent methyl halide transferase (MHT) and over-expressing one or more of a methyl formate synthase, a methanol dehydrogenase, and a hydrolytic dehydrogenase.
17. A bacteria-yeast co-culture comprising:
(i) bacteria which metabolize cellulose and produce one or more metabolic products, and
(ii) a yeast according to claim 15 , wherein the yeast uses at least one metabolic product produced by the bacteria as a carbon source.
18. The co-culture system of claim 17 wherein the yeast is selected from the group consisting of Saccharomyces, Pichia, Hansenula, Kluyveromyces, Yarrowia, Trichoderma and Scizosacchromyces.
19. The co-culture of claim 17 in which the yeast is S. cerevisiae and the bacterium is Actinotalea fermentans.