The oleaginous yeast Yarrowia lipolytica shows high potential as an isoprenoid platform. Considered
a GRAS microorganism, it has a high cytosolic acetyl-CoA
ux which is the main precursor of the
Mevalonate pathway for the synthesis of isoprenoids.
In this study we have engineered Y. lipolytica with the help of CRISPR/Cas9 to ...»»»»
The oleaginous yeast Yarrowia lipolytica shows high potential as an isoprenoid platform. Considered
a GRAS microorganism, it has a high cytosolic acetyl-CoA
ux which is the main precursor of the
Mevalonate pathway for the synthesis of isoprenoids.
In this study we have engineered Y. lipolytica with the help of CRISPR/Cas9 to optimize the Mevalonate
pathway in order to increase the production of the isoprenoids precursors. We have constructed
several strains for the production of limonene (monoterpene), squalene (triterpene), 2,3-oxidosqualene
(triterpene), valencene (sesquiterpene), -farnesene (sesquiterpene), ent-kaurine (diterpene) and -
Carotene (tetraterpene). Biosynthesis was con rmed for all of them but only -farnesene, squalene
and 2,3-oxidosqualene could be tested for production due to a lack of time.
It was possible to prove that Yarrowia lipolytica can be used for the production of several isoprenoids.
Improvement of the Mevalonate pathway has shown to have positive e ects in all tested production
strains. We succeeded in cloning the di erent genes which encode for the speci c synthases for each
product. We identi ed speci c changes in the precursor pathway of squalene and 2,3 oxidosqualene
that are bene cial for its production. Obtained concentrations for squalene are competitive compared
to the literature for Y. lipolytica.^^^^
El llevat oleagin os Yarrowia lipolytica mostra un gran potencial com a organisme plataforma per als
isoprenoides. Considerat un organisme GRAS, t e un gran
ux d'Acetil-CoA citoplasm atic el qual es
el precursor principal de la ruta del Mevalonat per a la s ntesi d'isoprenoides.
En aquest estudi hem fet enginyeria gen etica ...»»»»
El llevat oleagin os Yarrowia lipolytica mostra un gran potencial com a organisme plataforma per als
isoprenoides. Considerat un organisme GRAS, t e un gran
ux d'Acetil-CoA citoplasm atic el qual es
el precursor principal de la ruta del Mevalonat per a la s ntesi d'isoprenoides.
En aquest estudi hem fet enginyeria gen etica sobre Y. lipolytica amb l'ajuda de CRISPR/Cas9
per optimitzar la ruta del Mevalonat amb l'objectiu d'augmentar la producci o de precursors dels
isoprenoides. Hem constru t diverses soques per a la producci o de limonina (monoterp e), esqual e
(triterp e), 2,3-oxidoesqual e (triterp e), valencina (sesquiterp e), -farnes e (sesquiterp e), ent-kaurina
(diterp e) i -Carot e (tetraterp e). La bios ntesi es va veri car per a totes les soques per o nom es el
-farnes e, l'esqual e i el 2,3-oxidoesqual e varen poder ser testejats per a la producci o per falta de temps.
Va ser possible provar que Yarrowia lipolytica pot ser empleat per a la producci o de diversos isoprenoides.
L'optimitzaci o de la ruta del Mevalonat ha mostrat efectes positius en totes les soques
testejades. Hem tingut exit en la clonaci o de diversos gens que codi quen per sintases espec ques per
a cada producte. Hem identi cat canvis espec cs en la via metab olica del precursor de l'esqual e i el
2,3-oxidoesqual e que s on bene cioses per a la seva producci o. Les concentracions d'esqual e obtingudes
s on competents si les comparem amb la literatura de Y. lipolytica.^^^^
