US 12,110,526 B2
Cell-free production of sugars
Daniel MacEachran, Medford, MA (US); Drew S. Cunningham, Winchester, MA (US); William Jeremy Blake, Winchester, MA (US); and Matthew Eduardo Moura, Cambridge, MA (US)
Assigned to GreenLight Biosciences, Inc., Medford, MA (US)
Filed by GreenLight Biosciences, Inc., Medford, MA (US)
Filed on Oct. 22, 2021, as Appl. No. 17/507,939.
Application 17/507,939 is a continuation of application No. 16/892,696, filed on Jun. 4, 2020, abandoned.
Application 16/892,696 is a continuation of application No. 16/745,164, filed on Jan. 16, 2020, granted, now 10,704,067.
Application 16/745,164 is a continuation of application No. 16/395,548, filed on Apr. 26, 2019, granted, now 10,577,635.
Application 16/395,548 is a continuation of application No. 16/033,317, filed on Jul. 12, 2018, granted, now 10,316,342.
Application 16/033,317 is a continuation of application No. PCT/US2018/012516, filed on Jan. 5, 2018.
Claims priority of provisional application 62/538,181, filed on Jul. 28, 2017.
Claims priority of provisional application 62/443,447, filed on Jan. 6, 2017.
Prior Publication US 2022/0282291 A1, Sep. 8, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 19/02 (2006.01); C12N 9/10 (2006.01); C12N 9/90 (2006.01); C12N 9/92 (2006.01); C12N 15/52 (2006.01); C12P 19/14 (2006.01)
CPC C12P 19/02 (2013.01) [C12N 9/1051 (2013.01); C12N 9/90 (2013.01); C12N 9/92 (2013.01); C12N 15/52 (2013.01); C12P 19/14 (2013.01); C12Y 204/01049 (2013.01); C12Y 503/01 (2013.01); C12Y 503/01009 (2013.01); C12Y 504/02002 (2013.01); C12Y 504/02005 (2013.01); C12Y 504/02006 (2013.01)] 20 Claims
 
1. 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.