US 11,781,122 B2
Ectoine-producing yeast
Dominique Louis, Forges les Bains (FR); Karine Jaillardon, Saint Michel sur Orge (FR); and Dominique Thomas, Gif sur Yvette (FR)
Assigned to GIVAUDAN SA, Vernier (CH)
Appl. No. 16/630,262
Filed by ALDERYS, Orsay (FR)
PCT Filed Jul. 10, 2018, PCT No. PCT/EP2018/068719
§ 371(c)(1), (2) Date Jan. 10, 2020,
PCT Pub. No. WO2019/011947, PCT Pub. Date Jan. 17, 2019.
Claims priority of application No. 17305910 (EP), filed on Jul. 11, 2017.
Prior Publication US 2021/0222136 A1, Jul. 22, 2021
Int. Cl. C12N 9/12 (2006.01); C12N 9/04 (2006.01); C12N 9/02 (2006.01); C12N 9/10 (2006.01); C12N 9/80 (2006.01); C12N 9/88 (2006.01); C12P 17/12 (2006.01)
CPC C12N 9/1217 (2013.01) [C12N 9/0006 (2013.01); C12N 9/0008 (2013.01); C12N 9/1029 (2013.01); C12N 9/1096 (2013.01); C12N 9/80 (2013.01); C12N 9/88 (2013.01); C12P 17/12 (2013.01); C12Y 101/01003 (2013.01); C12Y 102/01011 (2013.01); C12Y 203/01031 (2013.01); C12Y 203/01178 (2013.01); C12Y 206/01076 (2013.01); C12Y 207/02004 (2013.01); C12Y 402/01108 (2013.01)] 15 Claims
 
1. An ectoine-producing recombinant yeast, in the genome of which:
(A) (i) at least one nucleic acid encoding an aspartokinase is overexpressed and/or is under the control of an inducible or repressible promoter; and/or
(ii) at least one nucleic acid encoding an aspartate kinase is overexpressed and/or is under the control of an inducible or repressible promoter; and
(B) at least one nucleic acid encoding an aspartate semi-aldehyde dehydrogenase that can use as coenzyme both NAD and NADP is overexpressed and/or is under the control of an inducible or repressible promoter; and
(C) at least one nucleic acid encoding a diaminobutyrate aminotransferase is overexpressed and/or is under the control of an inducible or repressible promoter; and
(D) (i) at least one nucleic acid encoding an homoserine-O-acetyltransferase METX is overexpressed and/or is under the control of an inducible or repressible promoter that is heterologous from the nucleic acid encoding METX, and
(ii) at least one nucleic acid encoding a diaminobutyric acid acetyltransferase is overexpressed and/or is under the control of an inducible or repressible promoter; and
(E) at least one nucleic acid encoding an ectoine synthase is overexpressed and/or is under the control of an inducible or repressible promoter; and
(F) (i) at least one endogenous nucleic acid encoding an homoserine dehydrogenase has been deleted and/or interrupted, and/or
(ii) at least one nucleic acid encoding a homoserine dehydrogenase is:
under the control of an inducible or repressible promoter;
under the control of a weak promoter; and/or
the homoserine dehydrogenase is degron-tagged.