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Table 5 Synthetic pathways based on isoprenoids, listed in chronological order

From: Yeast cell factories for fine chemical and API production

Host organism

Engineering steps

Substrate

Product/Outcome

Ref

Saccharomyces cerevisiae

1) Introduction of the Erwinia uredovora carotenoid biosynthesis genes [216]crtE, crtB, crtI and crtY (→ β-carotene) and crtE, crtB and crtI (→ lycopene), respectively, under the control of S. cerevisiae promoters and terminators [221]

galactose

β-Carotene: 0.103 mg/g [CDW]

 

[221–223]

 

2) Introduction of Erwinia herbicola carotenoid biosynthesis genes for lycopene, β-carotene and zeaxanthin production under the control of S. cerevisiae promoters and terminators [222]

 

Lycopene: 0.113 mg/g [CDW]

 

[221–223]

   

Zeaxanthin: 0.01% of CDW ~0.2 – 0.05 mg/g [CDW]

 

[222, 223]

Candida utilis

1) Introduction of synthetic, codon-optimized Erwinia uredovora carotenoid biosynthesis genes (crtE, crtB, crtI and crtY) [216] and Agrobacterium aurantiacum carotenoid biosynthesis genes (crtZ and crtW) [217] under the control ot C. utilis promoters and terminators:

glucose

Astaxanthin: 0.4 mg/g [CDW]

 

[219]

 

- Astaxanthin: crtE, crtB, crtI, crtY, crtZ and crtW

 

β-Carotene: 0.4 mg/g [CDW]

 

[219]

 

- β-Carotene: crtE, crtB, crtI and crtY

    
 

- Lycopene: crtE, crtB and crtI

    
 

2) Improving Lycopene yields [218] by disruption of the C. utilis squalene synthase gene (ERG9) and overex-pression of the catalytic domain of the C. utilis 3-hydroxy methylglutaryl CoA reductase gene (HMG)

 

Lycopene: 1.1 mg/g [CDW][219]

7.8 mg/g [CDW][218]

 

[218, 219]

Saccharomyces cerevisiae (fen1)

1) Transfer of expression cassettes for mature bovine adrenodoxin (ADX), adrenodoxin reductase (ADR), and side chain cleavage cytochrome P450 (P450scc)

galactose

60 mg/L

[241]

 

2) Transfer of Arabidopsis thaliana Δ7-sterol reductase

    
 

3) Disruption of Δ22-sterol desaturase (one step of endogenous ergosterol biosynthetic pathway)

    

Saccharomyces cerevisiae (fen1)

1) Transfer of expression cassettes for mature bovine adrenodoxin (ADX), adrenodoxin reductase (ADR), and side chain cleavage cytochrome P450 (P450scc)

galactose

No value given

[241]

 

2) Transfer of Arabidopsis thaliana Δ7-sterol reductase

    
 

3) Disruption of Δ22-sterol desaturase (one step of endogenous ergosterol biosynthetic pathway)

    
 

4) Introduction of type II human 3β-hydroxy-steroid dehydrogenase-isomerase (3β-HSD)

    

Saccharomyces cerevisiae

1) Rerouting the ergosterol biosynthesis pathway

glucose/ethanol

11.5 mg/L

[228]

 

2) Introduction of the mammalian-specific part of the hydrocortisone biosynthetic pathway

    
 

3) Inactivation of side reactions to steroid biosynthesis dead ends

    
 

4) Adjusting expression levels for optimized steroid channeling to hydrocortisone

    

Saccharomyces cerevisiae

1) Introduction of the Artemisia annua epicedrol synthase gene

galactose

0.37 mg/L

[229]

 

2) Overexpression of a truncated Hydroxy-methylglutaryl CoA reductase (trHmg1p)

    
 

3) Mutation of the Upc2p transcription factor → introduction of the upc2-1 allele with G888D [242]

    
 

4) Employing the S.c. haploid mating type a

    

Saccharomyces cerevisiae

1) Introduction of five Taxol biosynthetic genes from Taxus species: geranylgeranyl disphosphate synthase (GGPPS), taxadiene synthase (TS), taxadiene 5α-hydroxylase (THY5a), taxadienol 5α-O-acetyl trans-ferase (TAT), taxoid 10β-hydroxylase (THY10b) with necessary modifications for the expression in S. c.

simple sugar (glucose, galactose) and [2-14C] mevalonic acid for radio-HPLC analysis

taxadien-5α-ol: 0.025 mg/L no taxadien-5α-acetoxy-10β-ol in vivo

 

[230]

 

2) Due to restricted THY5a expression, only a very small amount of the intermediate taxadien-5α-ol and no taxadien-5α-acetoxy-10β-ol was detected in vivo [230]

    

Saccharomyces cerevisiae

1) Engineering the farnesyl pyrophosphate (FPP) biosynthetic pathway

simple sugar

~32 mg/L

[226]

 

2) Introduction of the Artemisia annua L amorphadiene synthase gene (FPP → amorphadiene)

    
 

3) Cloning the A. annua CYP71AV1/CPR (3-step oxidation: amorphadiene → artemisinic acid)

    

Saccharomyces cerevisiae

1) Follow-up study of [226]:

glucose

~380 mg/L

~120 mg/L

 

[227]

 

2) Engineering the pyruvate dehydrogenase bypass (pyruvate to acetyl-CoA) by overexpression of

    
 

- Salmonella acetyl-CoA synthetase variant (L641P)

    
 

- S. cerevisiae cytosolic acetaldehyde dehydrogenase (ALD6)

    
 

- In strain S. cerevisiae EPY224 [226]

    
 

3) Results: increased levels of mevalonate and amorpha-4,11-diene (~120 mg/L); generally applicable for isoprenoid production

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