US 11,987,829 B2
Method for the biosynthesis of diosmin and/or hesperidin in a microorganism
Cyrille Pauthenier, Juvisy-sur-Orge (FR); André Le Jeune, Draveil (FR); Hélène Scornec, Longjumeau (FR); Célia Roussel, Saintry sur Seine (FR); and Laetitia Joubert, Palaiseau (FR)
Assigned to LES LABORATOIRES SERVIER, Suresnes (FR)
Appl. No. 17/429,459
Filed by LES LABORATOIRES SERVIER, Suresnes (FR)
PCT Filed Feb. 11, 2020, PCT No. PCT/EP2020/053503
§ 371(c)(1), (2) Date Aug. 9, 2021,
PCT Pub. No. WO2020/165189, PCT Pub. Date Aug. 20, 2020.
Claims priority of application No. 19305163 (EP), filed on Feb. 11, 2019.
Prior Publication US 2022/0042061 A1, Feb. 10, 2022
Int. Cl. C12P 19/60 (2006.01); C12N 9/00 (2006.01); C12N 9/02 (2006.01); C12N 9/10 (2006.01); C12N 9/88 (2006.01); C12N 9/90 (2006.01)
CPC C12P 19/60 (2013.01) [C12N 9/0042 (2013.01); C12N 9/0071 (2013.01); C12N 9/0073 (2013.01); C12N 9/1007 (2013.01); C12N 9/1051 (2013.01); C12N 9/1085 (2013.01); C12N 9/88 (2013.01); C12N 9/90 (2013.01); C12N 9/93 (2013.01); C12Y 106/02004 (2013.01); C12Y 114/11022 (2013.01); C12Y 114/13011 (2013.01); C12Y 114/14 (2013.01); C12Y 204/01 (2013.01); C12Y 204/01185 (2013.01); C12Y 205/01006 (2013.01); C12Y 402/01076 (2013.01); C12Y 403/01023 (2013.01); C12Y 403/01024 (2013.01); C12Y 505/01006 (2013.01); C12Y 602/01012 (2013.01)] 18 Claims
 
1. A recombinant microorganism comprising:
a heterologous nucleic acid sequence coding for a flavanone 7-O-beta-D-glucosyltransferase (UGT) belonging to EC 2.4.1.185, which is capable of adding a glucose in position 7 of hesperetin and/or diosmetin; and
a heterologous nucleic acid sequence coding for a 6″-O-rhamnosyltransferase (RhaT) belonging to EC 2.4.1.653, which is capable of transferring a rhamnose into position 6 of the glucose of hesperetin-7-O-glucoside and/or diosmetin-7-O-glucoside; and
a heterologous nucleic acid sequence coding for a UDP-glucose 4,6-dehydratase/UDP-4-keto-6-deoxy-D-glucose 3,5-epimerase/UDP-4-keto-L-rhamnose-reductase (RHM) belonging to EC 4.2.1.76, which is capable of producing UDP-rhamnose.