Metabolic engineering of Yarrowia lipolytica for terpenoids production: advances and perspectives

Figures & data

Figure 1. The biosynthetic pathways of terpenoids. (A) MVA pathway. (B) The decoration process of representative terpenes. (C) Noncanonical precursors derived from DMAPPs. Metabolites: HMG-CoA, 3-hydroxy-3-methylglutaryl-CoA; IPP, isopentenyl diphosphate; DMAPP, dimethylallyl diphosphate; NPP, neryl diphosphate; GPP, geranyl diphosphate; FPP, farnesyl diphosphate; GGPP, geranylgeranyl diphosphate; CPP, chrysanthemyl diphosphate; LPP, lavandulyl diphosphate; MPP, maconellyl diphosphate; PPP, planococcyl diphosphate. Enzymes: ERG10, acetyl-CoA acetyltransferase; ERG13, HMG-CoA synthase; HMGR, HMG-CoA reductase; ERG12, mevalonate kinase; ERG8, phosphomevalonate kinase; ERG19, mevalonate diphosphate decarboxylase; IDI, IPP isomerase; ERG20, farnesyl pyrophosphate synthetase; ERG9, squalene synthase; ERG1, squalene epoxidase; ERG7, lanosterol synthase; NDPS1, neryl diphosphate synthase; PS, pinene synthase; LS, limonene synthase; LIS, linalool synthase; FS, α-farnesene synthase; BFS, β-farnesene synthase; STS, α-santalene synthase; ADS, amorphadiene synthase; BS, bisabolene synthase; BBS, (−)-α-bisabolol synthase; VS, valencene synthase; CYP706M1, (+)-nootkatone synthase; bAS, β-amyrin synthase; CYP716A12, oleanolic acid synthase; LUS, lupeol synthase; CYP716A180, betulinic acid synthase; DS, dammarenediol-II synthase; PPDS, protopanaxadiol synthase; UGT1, UDP-glucose glucosyltransferase; CCD1, carotenoid cleavage dioxygenase; GGPPS, geranylgeranyl diphosphate synthase; CarRP, phytoene synthase/lycopene cyclase; CarB, phytoene dehydrogenase; CrtW, β-carotene ketolase; CrtZ, β-carotene hydrolase.

Table 1. Summary of metabolic engineering strategies for improving terpenoids biosynthesis in Y. lipolytica, and information of representative terpenes with the highest titers reported in E. coli and S. cerevisiae.

Products	Strains	Engineering strategies	Medium/ Carbon source	Fermentation	Titer	References
Monoterpenoids
Limonene	Y. lipolytica Po1f	HMGR, ERG12, tSlNDPS1, tArLS	YPD Glucose Pyruvate	Flask Batch	23.56 mg/L	[32]
Limonene	Y. lipolytica Po1f	HMGR, ERG12, tSlNDPS1, tArLS	YPG Glycerol Citrate	1.5-L fermenter Fed-batch	165.3 mg/L	[33]
Limonene	Y. lipolytica Po1g	Δku70, HMGR, l-MsLS, d-ClLS	YPDM Glucose	5-L fermenter Fed-batch	L-limonene11.705 mg/L D-limonene11.088 mg/L	[34]
Limonene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, ERG20^88W-N119W, PERG11-ERG9, PfLS	YPD Glucose	Glass tube Batch	35.9 mg/L	[28]
Limonene	Y. lipolytica Po1f	SsXR, SsXDH, XK, HMGR, ERG12, tSlNDPS1, tArLS,	YP Lignocellulose hydrolysate	Flask Batch	20.57 mg/L	[35]
Linalool	Y. lipolytica Po1f	HMGR, IDI, ERG20^88W-N119W, AaLIS	YP Citrate Pyruvate	Flask Batch	6.96 mg/L	[36]
α-Pinene	Y. lipolytica Po1f	AMPD, HMGR, ERG12, ERG8, tSlNDPS1, PtPS, tPtPS	YP Lignocellulosic hydrolysate	Flask Batch	36.1 mg/L	[37]
Limonene	E. coli BL21 (DE3)	atoB, ScERG13, tScHMG1, ScERG12, ScERG8, ScERG19, IDI, tAgGPPS, tMsLS	M9 Glycerol	3.1-L fermenter Fed-batch	3.63 g/L	[38]
Limonene	S. cerevisiae EGY48	EfmvaS, EfmvaE, ERG12, ERG8, ERG19, IDI, ERG20^N127W, ClLS Compartmentalization	SD Galactose Raffinose	Flask Fed-batch	2.58 g/L	[39]
Sesquiterpenoids
(+)-Nootkatone	Y. lipolytica ATCC 201249	tScHMG1, ERG20, CnVS, CnCYP706M1-tAtCPR1	YPD Glucose	Flask Batch	0.9782 mg/L	[40]
α-Santalene	Y. lipolytica ATCC 201249	ScHMG1, ERG8, ClSTS	YPD Glucose	5-L fermenter Fed-batch	27.92 mg/L	[41]
Valencene	Y. lipolytica ATCC 201249	tScHMG1, ERG20, CnVS, CnCYP706M1-tAtCPR1	YPD Glucose	Flask Batch	22.8 mg/L	[40]
Valencene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, ERG20, PERG11-ERG9, CnVS	YPD Glucose	Glass tube Batch	113.9 mg/L	[28]
Amorphadiene	Y. lipolytica Po1g	HMGR, ERG12, AaADS Supplementation of cerulenin	Complete synthetic media Glucose	Flask Batch	171.5 mg/L	[42]
α-Farnesene	Y. lipolytica Po1h	tScHMG1, IDI, MdFS-linker-ERG20	PM Glucose Fructose	1.5-L fermenter Batch	259.98 mg/L	[43]
α-Farnesene	Y. lipolytica Po1f	EcatoB, ERG13, BpHMGR, ERG12, ERG8, ERG19, IDI, GPPS, MdFS-linker-ERG20	YPD Glucose	1-L fermenter Fed-batch	25.55 g/L	[44]
β-Farnesene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, ERG20, AaBFS	YPD Glucose	Glass tube Batch	955 mg/L	[28]
Bisabolene	Y. lipolytica Po1g	Δku70, HMGR, GcABC-G1, α-AgBS, β-ZoBS, γ-HaBS	YP with Mg^2+ WCO	Flask Batch	α-bisabolene 973.1 mg/L β-bisabolene 68.2 mg/L γ-bisabolene 20.2 mg/L	[45]
(−)-α-Bisabolol	Y. lipolytica Po1f	Δku70, tHMGR, ERG20, PERG9_50bp-ERG9, POT1, MrBBS	YPD Glucose	Flask Batch	364.23 mg/L	[46]
β-Farnesene	E. coli BL21 (DE3)	atoB, ScERG13, tScHMG1, ScERG12, ScERG8, ScERG19, IDI, ispA, AaFS	2 × YT Glycerol	7-L fermenter Batch	8.74 g/L	[47]
β-Farnesene	S. cerevisiae Y227	Δgal80, Δhxt3, Δdit1, Δndt80, Δho, Δgal2, Δadh5, Δald4, Δald6, Δrhr2, Δygr250c, Δbdh1, Δadh1, GAL4, GAL4*, ERG10, ERG13, ERG13*, tHMG1, ERG12, ERG8, ERG19, ERG20, PMET3-ERG9, PDR3, BjHMGs, CaTHL, AaFS, ACS2, ZmPDC, DzeutE, LmPK, CkPTA, SpHMGr	Mineral salt Cane sirup	0.5-L fermenter Fed-batch	130 g/L	[48]
Triterpenoids
Protopanaxadiol	Y. lipolytica ATCC 201249	Δku70, Δpox1-3, TX, SsXR, SsXDH, XKS, TKL, TAL, tHMGR, ERG20, ERG9, PgDS, PgPPDS-linker-AtATR1	YPX Xylose	5-L fermenter Fed-batch	300.63 mg/L	[49]
Ginsenoside compound K	Y. lipolytica ATCC 201249	tHMGR, ERG20, ERG9, PgDS, PgPPDS-linker-AtATR1, PgUGT1	YPD Glucose	5-L fermenter Fed-batch	161.8 mg/L	[50]
2,3-Oxidosqualene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, ERG20, ERG9, ERG1, PERG7_50bp-ERG7	YPD Glucose	Deepwell plate Batch	22 mg/L	[28]
Oleanolic acid	Y. lipolytica ATCC 201249	tHMGR, ERG20, ERG9, GgbAS, MtCYP716A12-linker-AtATR1	YPD Glucose	5-L fermenter Fed-batch	540.7 mg/L	[51]
Lupeol	Y. lipolytica ATCC 201249	Δpah1, Δdgk1, HMGR, ERG9, ERG1, OLE1, RcLUS	YPD Glucose	Flask Batch	411.72 mg/L	[52]
Betulinic acid	Y. lipolytica ATCC 201249	tHMGR, ERG9, AtLUP1, MtCYP716A12-AtCPR1	YPG Glycerol	Flask Batch	26.53 mg/L	[53]
Betulinic acid	Y. lipolytica ATCC 201249	Δku70, Δku80, MFE1, HMGR, ERG9, ERG1, LjCPR, BPLO, RcLUS	YPD Glucose	Flask Batch	204.89 mg/L	[54]
Squalene	Y. lipolytica Po1f	ACL1, SeACS^L641P, HMGR	YPD Glucose Citrate	Flask Batch	10 mg/g DCW	[55]
Squalene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, ERG20, ERG9, PERG7_50bp-ERG7	YPD Glucose	Deepwell plate Batch	402.4 mg/L	[28]
Squalene	Y. lipolytica Po1g	MnDH2, HMGR, ERG9	Minimal media Glucose	Flask Fed-batch	502.75 mg/L	[56]
Squalene	E. coli DH5α	SpmvaK1, SpmvaD, SpmvaK2, IDI, EfmvaE, EfmvaS, ispA, tScERG9, tsr	2 × YT Glycerol	Flask Batch	612 mg/L	[57]
Squalene	S. cerevisiae Y2805	tHMG1, EcispA Supplementation of terbinafine	YPD Glucose	5-L fermenter Fed-batch	2.011 g/L	[58]
Tetraterpenoids
Lycopene	Y. lipolytica Po1f	PaCrtE, PaCrtB, PaCrtI flux balance analysis	Designed-medium	1.5-L fermenter Fed-batch	242 mg/L	[59]
Lycopene	Y. lipolytica Po1f	HMGR, EGR8, ERG19, PaCrtB, PaCrtE. PaCrtI	YPD Glucose	1-L fermenter Fed-batch	213 mg/L	[60]
Lycopene	Y. lipolytica ATCC 201249	ScERG20, ScERG8, ScIDI, POT1, EnCrtE, EnCrtB, EnCrtI,	SC Glucose	Flask Batch	259 mg/L	[61]
Lycopene	Y. lipolytica Po1f	AMPD, PaCrtE, PaCrtB, PaCrtI	MM Glucose	5-L fermenter Fed-batch	745 mg/L	[62]
Lycopene	Y. lipolytica Po1f	AtIPK, ScCHK, ERG20, PvIDI, LpCrtE, LpCrtB, LpCrtI	YNB Glucose Palmitic acid Isoprenol	3-L fermenter Batch	4.2 g/L	[63]
β-Carotene	Y. lipolytica Po1f	Δku70, Δku80, GGPPS, SsCarS	YPD Glucose	Flask Batch	0.41 mg/g DCW	[64]
β-Carotene	Y. lipolytica Po1f	Δku70, Δpox3-6, Δlip1, ΔYALI0F30987g, ERG10, ERG13, tHMGR, ERG12, ERG8, ERG19, IDI, ERG20, GGPPS, McCarRP, McCarB	YPD Glucose	1.5-L fermenter Fed-batch	4.00 g/L	[65]
β-Carotene	Y. lipolytica Po1d	Δpox1-6, Δtgl4, DGA2, GPD1, HMGR, GGPPS, McCarRP, McCarB	YPD Glucose	5-L fermenter Fed-batch	6.5 g/L	[66]
β-Carotene	Y. lipolytica ST6512	ACL1, SeACS, HMGR, ERG12, IDI, GGPPS, PERG11-ERG9, XdCrtYB, XdCrtI	YPD Glucose	Deepwell plate Batch	164 mg/L	[28]
β-Carotene	Y. lipolytica Po1f	Δmfe, ΔPox2-3, Δlip1 ERG13, HMGR, GGPPS, McCarRP, McCarB	YPD Glucose	5-L fermenter Fed-batch	4.5 g/L	[67]
β-Carotene	Y. lipolytica Po1f	Δku70, Δsnf, HXK, ERG13, tHMGR, GGPPS, BtCarRA, BtCarB	YPD Glucose	50-L fermenter Fed-batch	2.4 g/L	[68]
Astaxanthin	Y. lipolytica GB20	HMGR, XdCrtE, XdCrtYB, XdCrtI, PsCrtW, PaCrtZ, PERG9_50bp-ERG9	YPD Glucose	24-well plates	54.6 mg/L	[69]
Astaxanthin	Y. lipolytica GB20	HMGR, SsGGPPS, XdCrtE, XdCrtYB, XdCrtI, HpBKT, HpCrtZ, PERG9_50bp-ERG9	YPD Glucose	1-L fermenter Fed-batch	285 mg/L	[70]
α-Ionone	Y. lipolytica CLIB138	SsNphT7, ERG13, tHMGR, ERG12, ERG8, ERG19, HpIDI, GPPS, ERG20-GGPPS, McCarRP*-McCarB, LsLCYe*, OfCCD1	YPD Glucose	5-L fermenter Fed-batch	408 mg/L	[71]
β-Ionone	Y. lipolytica CLIB138	SsNphT7, ERG13, tHMGR, ERG12, ERG8, ERG9, HpIDI, GPPS, ERG20-GGPPS, McCarRP-McCarB, OfCCD1	YPD Glucose	2-L fermenter Fed-batch	380 mg/L	[72]
β-Ionone	Y. lipolytica Po1f	Δku70, Δku80, Δpox5, Δpox3, Δlip1, ΔD17, ERG10, ERG13, tHMGR, ERG12, ERG8, ERG19, IDI, ERG20, GGPPS, BbPK, BsPTA, McCarB, McCarRP, PhCCD1	YPD Glucose	3-L fermenter Fed-batch	0.98 g/L	[73]
β-Carotene	E. coli BL21 (DE3)	ScERG12, ScERG8, ScERG19, ScIDI, EfmvaE, EfmvaS, Bsdxs, Bsfni, AgGPPS2, EhCrtE, EhCrtB, EhCrtI, EhCrtY	Mineral salt with beef extract Glucose Glycerol	5-L fermenter Fed-batch	3.2 g/L	[74]
β-Carotene	S. cerevisiae D452-2	Δald6, SsXYL1, SsXYL2 XKS1, XdCrtYB, XdCrtI, XdCrtE*	Verduyn medium Xylose	3-L fermenter Fed-batch	772.8 mg/L	[75]

“Δ” refers to gene knockouts; “*” refers to gene with mutant; “-” refers to protein fusion, “P_XX-XX” refers to genes drived by specific promoters. The uppercase and lowercase letters before some genes refer to the source of the gene. The information about terpenes produced in E. coli and S. cerevisiae are marked with blue shading.

Figure 2. Overview of the engineering strategies for terpenoid production in Y. lipolytica. (A) Engineering substrate utilization to minimize production costs. HXK, hexokinase; SsXR, xylose reductase from Scheffersomyces stipitis; SsXDH, xylitol dehydrogenase from Scheffersomyces stipitis; XKS, xylulose kinase. (B) Increasing cytosolic acetyl-CoA supply by introducing heterologous pathways or redirecting native central carbon metabolism. AMPD, AMP deaminase; ICDH, isocitrate dehydrogenase; BbPK, phosphoketolase from Bifidobacterium bifidum; BsPTA, phosphotransacetylase from Bacillus subtilis; ACL, ATP citrate lyase; SeACS, acetyl-CoA synthetase from Salmonella enterica; MFE1, multifunctional β-oxidation enzyme. (C) Overexpressing the genes in MVA pathway and engineering cofactor supply. GND2, 6-phosphogluconate dehydrogenase; MnDH, mannitol dehydrogenase; MAE1, malic enzyme; UGA2, succinate semialdehyde dehydrogenase; IDP2, cytosolic NADP⁺-specific isocitrate dehydrogenase; AtIPK, isopentenyl phosphate kinase from Arabidopsis thaliana; ScCHK, bifunctional choline kinase/ethanolamine kinase from Saccharomyces cerevisiae. (D) Fine-tuning the expression of key enzymes: combinatorial optimization of P450s-CPRs among species and constructing self‐sufficient P450s by fusion with CPRs; protein engineering to improve the specificity and activity of enzymes; relieving side pathway competition. (E) Modulating hydrophobicity for terpenoid accumulation: increasing intracellular lipids by genetic manipulation or feeding with hydrophobic substrates and in situ trapping with organic overlay. (F) Engineering based on metabolomics analysis: modeling-guided fermentation optimization and metabolic engineering. FBA: flux balance analysis.

Cao X, Lv YB, Chen J, et al. Metabolic engineering of oleaginous yeast Yarrowia lipolytica for limonene overproduction. Biotechnol Biofuels. 2016;9:214.

 PubMed Web of Science ®Google Scholar

Cheng BQ, Wei LJ, Lv YB, et al. Elevating limonene production in oleaginous yeast Yarrowia lipolytica via genetic engineering of limonene biosynthesis pathway and optimization of medium composition. Biotechnol Bioproc E. 2019;24(3):500–506.

 Web of Science ®Google Scholar

Pang YR, Zhao YK, Li SL, et al. Engineering the oleaginous yeast Yarrowia lipolytica to produce limonene from waste cooking oil. Biotechnol Biofuels. 2019;12:241.

 PubMed Web of Science ®Google Scholar

Arnesen JA, Kildegaard KR, Pastor MC, et al. Yarrowia lipolytica strains engineered for the production of terpenoids. Front Bioeng Biotechnol. 2020;8:945.

 PubMed Web of Science ®Google Scholar

Yao F, Liu SC, Wang DN, et al. Engineering oleaginous yeast Yarrowia lipolytica for enhanced limonene production from xylose and lignocellulosic hydrolysate. FEMS Yeast Res. 2020;20(6):foaa046.

 PubMed Web of Science ®Google Scholar

Cao X, Wei LJ, Lin JY, et al. Enhancing linalool production by engineering oleaginous yeast Yarrowia lipolytica. Bioresour Technol. 2017;245(Pt B):1641–1644.

 PubMedGoogle Scholar

Wei LJ, Zhong YT, Nie MY, et al. Biosynthesis of α-pinene by genetically engineered Yarrowia lipolytica from low-cost renewable feedstocks. J Agric Food Chem. 2021;69(1):275–285.

 PubMed Web of Science ®Google Scholar

Rolf J, Julsing MK, Rosenthal K, et al. A gram-scale limonene production process with engineered Escherichia coli. Molecules. 2020;25(8):1881.

 Web of Science ®Google Scholar

Dusseaux S, Wajn WT, Liu YX, et al. Transforming yeast peroxisomes into microfactories for the efficient production of high-value isoprenoids. Proc Natl Acad Sci U S A. 2020;117(50):31789–31799.

 PubMed Web of Science ®Google Scholar

Guo XY, Sun J, Li DS, et al. Heterologous biosynthesis of (+)-nootkatone in unconventional yeast Yarrowia lipolytica. Biochem Eng J. 2018;137:125–131.

 Web of Science ®Google Scholar

Jia D, Xu S, Sun J, et al. Yarrowia lipolytica construction for heterologous synthesis of α-santalene and fermentation optimization. Appl Microbiol Biotechnol. 2019;103(8):3511–3520.

 PubMed Web of Science ®Google Scholar

Marsafari M, Xu P, Debottlenecking mevalonate pathway for antimalarial drug precursor amorphadiene biosynthesis in Yarrowia lipolytica. Metab Eng Commun. 2020;10:e00121.

 PubMedGoogle Scholar

Yang X, Nambou K, Wei LJ, et al. Heterologous production of α-farnesene in metabolically engineered strains of Yarrowia lipolytica. Bioresour Technol. 2016;216:1040–1048.

 PubMed Web of Science ®Google Scholar

Liu YH, Jiang X, Cui ZY, et al. Engineering the oleaginous yeast Yarrowia lipolytica for production of α-farnesene. Biotechnol Biofuels. 2019;12:296.

 PubMed Web of Science ®Google Scholar

Zhao YK, Zhu K, Li J, et al. High-efficiency production of bisabolene from waste cooking oil by metabolically engineered Yarrowia lipolytica. Microb Biotechnol. 2021:1–17.

 Web of Science ®Google Scholar

Ma YR, Li WJ, Mai J, et al. Engineering Yarrowia lipolytica for sustainable production of the chamomile sesquiterpene (−)-α-bisabolol. Green Chem. 2021;23(2):780–787.

 Web of Science ®Google Scholar

You SP, Yin QD, Zhang JY, et al. Utilization of biodiesel by-product as substrate for high-production of β-farnesene via relatively balanced mevalonate pathway in Escherichia coli. Bioresour Technol. 2017;243:228–236.

 PubMed Web of Science ®Google Scholar

Meadows AL, Hawkins KM, Tsegaye Y, et al. Rewriting yeast central carbon metabolism for industrial isoprenoid production. Nature. 2016;537(7622):694–697.

 PubMed Web of Science ®Google Scholar

Wu YF, Xu S, Gao X, et al. Enhanced protopanaxadiol production from xylose by engineered Yarrowia lipolytica. Microb Cell Fact. 2019;18(1):83.

 PubMedGoogle Scholar

Li DS, Wu YF, Zhang CB, et al. Production of triterpene ginsenoside compound K in the non-conventional yeast Yarrowia lipolytica. J Agric Food Chem. 2019;67(9):2581–2588.

 PubMed Web of Science ®Google Scholar

Li DS, Wu YF, Wei PP, et al. Metabolic engineering of Yarrowia lipolytica for heterologous oleanolic acid production. Cheml Eng Sci. 2020;218:115529.

 Web of Science ®Google Scholar

Zhang JL, Bai QY, Peng YZ, et al. High production of triterpenoids in Yarrowia lipolytica through manipulation of lipid components. Biotechnol Biofuels. 2020;13:133.

 PubMed Web of Science ®Google Scholar

Sun J, Zhang CB, Nan WH, et al. Glycerol improves heterologous biosynthesis of betulinic acid in engineered Yarrowia lipolytica. Chem Eng Sci. 2019;196:82–90.

 Web of Science ®Google Scholar

Jin CC, Zhang JL, Song H, et al. Boosting the biosynthesis of betulinic acid and related triterpenoids in Yarrowia lipolytica via multimodular metabolic engineering. Microb Cell Fact. 2019;18(1):77.

 PubMedGoogle Scholar

Huang YY, Jian XX, Lv YB, et al. Enhanced squalene biosynthesis in Yarrowia lipolytica based on metabolically engineered acetyl-CoA metabolism. J Biotechnol. 2018;281:106–114.

 PubMed Web of Science ®Google Scholar

Liu H, Wang F, Deng L, et al. Genetic and bioprocess engineering to improve squalene production in Yarrowia lipolytica. Bioresour Technol. 2020;317:123991.

 PubMed Web of Science ®Google Scholar

Meng YH, Shao XX, Wang Y, et al. Extension of cell membrane boosting squalene production in the engineered Escherichia coli. Biotechnol Bioeng. 2020;117(11):3499–3507.

 PubMedGoogle Scholar

Han JY, Seo SH, Song JM, et al. High-level recombinant production of squalene using selected Saccharomyces cerevisiae strains. J Ind Microbiol Biotechnol. 2018;45(4):239–251.

 PubMed Web of Science ®Google Scholar

Nambou K, Jian XX, Zhang XK, et al. Flux balance analysis inspired bioprocess upgrading for lycopene production by a metabolically engineered strain of Yarrowia lipolytica. Metabolites. 2015;5(4):794–813.

 PubMed Web of Science ®Google Scholar

Schwartz C, Frogue K, Misa J, et al. Host and pathway engineering for enhanced lycopene biosynthesis in Yarrowia lipolytica. Front Microbiol. 2017;8:2233.

 PubMed Web of Science ®Google Scholar

Liu D, Liu H, Qi H, et al. Constructing yeast chimeric pathways to boost lipophilic terpene synthesis. ACS Synth Biol. 2019;8(4):724–733.

 PubMed Web of Science ®Google Scholar

Zhang XK, Nie MY, Chen J, et al. Multicopy integrants of crt genes and co-expression of AMP deaminase improve lycopene production in Yarrowia lipolytica. J Biotechnol. 2019;289:46–54.

 PubMed Web of Science ®Google Scholar

Luo ZS, Liu N, Lazar Z, et al. Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity. Metab Eng. 2020;61:344–351.

 PubMed Web of Science ®Google Scholar

Gao SL, Tong YY, Zhu L, et al. Production of β-carotene by expressing a heterologous multifunctional carotene synthase in Yarrowia lipolytica. Biotechnol Lett. 2017;39(6):921–927.

 PubMed Web of Science ®Google Scholar

Gao SL, Tong YY, Zhu L, et al. Iterative integration of multiple-copy pathway genes in Yarrowia lipolytica for heterologous β-carotene production. Metab Eng. 2017;41:192–201.

 PubMed Web of Science ®Google Scholar

Larroude M, Celinska E, Back A, et al. A synthetic biology approach to transform Yarrowia lipolytica into a competitive biotechnological producer of β-carotene. Biotechnol Bioeng. 2018;115(2):464–472.

 PubMed Web of Science ®Google Scholar

Zhang XK, Wang DN, Chen J, et al. Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica. Biotechnol Lett. 2020;42(6):945–956.

 PubMed Web of Science ®Google Scholar

Qiang S, Wang J, Xiong XC, et al. Promoting the synthesis of precursor substances by overexpressing hexokinase (Hxk) and hydroxymethylglutaryl-CoA synthase (Erg13) to elevate β-carotene production in engineered Yarrowia lipolytica. Front Microbiol. 2020;11:1346.

 PubMed Web of Science ®Google Scholar

Kildegaard KR, Adiego-Perez B, Belda DD, et al. Engineering of Yarrowia lipolytica for production of astaxanthin. Synth Syst Biotechnol. 2017;2(4):287–294.

 PubMed Web of Science ®Google Scholar

Tramontin LRR, Kildegaard KR, Sudarsan S, et al. Enhancement of astaxanthin biosynthesis in oleaginous yeast Yarrowia lipolytica via microalgal pathway. Microorganisms. 2019;7(10):472.

 Web of Science ®Google Scholar

Czajka JJ, Kambhampati S, Tang YJ, et al. Application of stable isotope tracing to elucidate metabolic dynamics during Yarrowia lipolytica α-ionone fermentation. iScience. 2020;23(2):100854.

 PubMed Web of Science ®Google Scholar

Czajka JJ, Nathenson JA, Benites VT, et al. Engineering the oleaginous yeast Yarrowia lipolytica to produce the aroma compound β-ionone. Microb Cell Fact. 2018;17(1):136.

 PubMedGoogle Scholar

Lu YP, Yang QY, Lin ZL, et al. A modular pathway engineering strategy for the high-level production of β-ionone in Yarrowia lipolytica. Microb Cell Fact. 2020;19(1):49.

 PubMed Web of Science ®Google Scholar

Yang JM, Guo LZ, Biosynthesis of β-carotene in engineered E. coli using the MEP and MVA pathways. Microb Cell Fact. 2014;13:160.

 PubMed Web of Science ®Google Scholar

Sun L, Atkinson CA, Lee YG, et al. High-level β-carotene production from xylose by engineered Saccharomyces cerevisiae without overexpression of a truncated HMG1 (tHMG1). Biotechnol Bioeng. 2020;117(11):3522–3532.

 PubMed Web of Science ®Google Scholar