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US 12,110,526 B2Granted
Cell-free production of sugars
AssigneeGreenLight Biosciences, Inc.
InventorsDaniel MacEachran, Drew S. Cunningham, William Jeremy Blake, Matthew Eduardo Moura
Life of this patent
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Abstract
Provided herein, in some embodiments, are systems, methods, and compositions (e.g., cells and cell lysates) for enzymatically converting a polymeric glucose carbohydrate (e.g., starch) to sugar.
Claims20
A method for producing allulose, the method comprising:
- (i) converting fructose 6-phosphate (F6P) to allulose-6-phosphate (A6P), catalyzed by an allulose 6-phosphate epimerase (A6PE) selected from the group consisting of Thermobacterium thermosaccharolyticum A6PE, Thermoanaerobacter brockii A6PE, Caldanaerobacter subterraneus A6PE, Deferribacter desulfuricans A6PE, Thermocrinis ruber A6PE, Hydrogenivirga sp. 128-5-R1-1 A6PE, Thermosipho atlanticus A6PE, and Thermosulfidibacter takaii A6PE;
- and
- (ii) converting A6P to allulose, catalyzed by an allulose 6-phosphate phosphatase (A6PP) selected from the group consisting of Thermoanaerobacter wiegelii A6PP, Thermoanaerobacter ethanolicus A6PP, Thermus islandicus A6PP, Deinococcus geothermalis A6PP, Thermosphaera aggregans A6PP, Crenarchaeota archaeon A6PP, Pyrococcus horikoshii A6PP, Aquifex aeolicus A6PP, Desulfotomaculum kuznetsovii A6PP, Caldanaerobacter subterraneus A6PP, Acidothermus cellulolyticus A6PP, Methanothermobacter thermautotrophicus A6PP, Thermobifida fusca A6PP, Thermotoga neapolitana A6PP, Petrotoga mobilis A6PP, Thermodesulfatator indicus A6PP, Thermus thermophilus A6PP, Bacteroides vulgatus A6PP, and Bacteroides fragilis A6PP.
A method for producing allulose, the method comprising:
- converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by an allulose 6-phosphate epimerase (A6PE), and
- converting A6P to allulose, catalyzed by an allulose 6-phosphate phosphatase (A6PP), in a cell lysate mixture comprising cell lysates from microbial cells that express one or both of the allulose 6-phosphate epimerase (A6PE) and the allulose 6-phosphate phosphatase (A6PP),
- wherein the A6PE is derived from an organism selected from the group consisting of Thermobacterium thermosaccharolyticum, Thermoanaerobacter brockii, Caldanaerobacter subterraneus, Deferribacter desulfuricans, Thermocrinis ruber, Hydrogenivirga sp. 128-5-R1-1, Thermosipho atlanticus, and Thermosulfidibacter takaii;
- and
- wherein the A6PP is derived from an organism selected from the group consisting of Thermoanaerobacter wiegelii, Thermoanaerobacter ethanolicus, Thermus islandicus, Deinococcus geothermalis, Thermosphaera aggregans, Crenarchaeota archaeon, Pyrococcus horikoshii, Aquifex aeolicus, Desulfotomaculum kuznetsovii, Caldanaerobacter subterraneus, Acidothermus cellulolyticus, Methanothermobacter thermautotrophicus, Thermobifida fusca, Thermotoga neapolitana, Petrotoga mobilis, Thermodesulfatator indicus, Thermus thermophilus, Bacteroides vulgatus, and Bacteroides fragilis.
2 more elements
A method for producing allulose, the method comprising:
- (i) converting fructose 6-phosphate (F6P) to allulose-6-phosphate (A6P), catalyzed by an allulose 6-phosphate epimerase (A6PE);
- and
- (ii) converting A6P to allulose, catalyzed by an allulose 6-phosphate phosphatase (A6PP), wherein:
- (a) the A6PE is selected from the group consisting of Thermobacterium thermosaccharolyticum A6PE, Thermoanaerobacter brockii A6PE, Caldanaerobacter subterraneus A6PE, Deferribacter desulfuricans A6PE, Thermocrinis ruber A6PE, Hydrogenivirga sp. 128-5-R1-1 A6PE, Thermosipho atlanticus A6PE, and Thermosulfidibacter takaii A6PE;
- and/or
- (b) the A6PP is selected from the group consisting of Thermoanaerobacter wiegelii A6PP, Thermoanaerobacter ethanolicus A6PP, Thermus islandicus A6PP, Deinococcus geothermalis A6PP, Thermosphaera aggregans A6PP, Crenarchaeota archaeon A6PP, Pyrococcus horikoshii A6PP, Aquifex aeolicus A6PP, Desulfotomaculum kuznetsovii A6PP, Caldanaerobacter subterraneus A6PP, Acidothermus cellulolyticus A6PP, Methanothermobacter thermautotrophicus A6PP, Thermobifida fusca A6PP, Thermotoga neapolitana A6PP, Petrotoga mobilis A6PP, Thermodesulfatator indicus A6PP, Thermus thermophilus A6PP, Bacteroides vulgatus A6PP, and Bacteroides fragilis A6PP.
3 more elements
Dependencies are inferred from the claim text.