Metabolic Engineering for Production of<i>β</i>-Carotene and Lycopene in<i>Saccharomyces cerevisiae</i>

Yamano, Shigeyuki; Ishii, Takeaki; Nakagawa, Masaya; Ikenaga, Hiroshi; Misawa, Norihiko

doi:10.1271/bbb.58.1112

Cited by 144 publications

(86 citation statements)

References 9 publications

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“…Accumulation of intermediates was not observed in the production of carotenoids in E. coli with the carotenogenic genes from E. uredovora (16). However, in S. cerevisiae cells overexpressing carotenogenic genes from E. uredovora on episomal vectors, intermediates also accumulated; 78% of the total carotenoids consisted of ␤-carotene, 11% accumulated as phytoene, and 11% accumulated as lycopene (41). Apparently, phytoene desaturation becomes a rate-limiting step in heterologous ␤-carotene production by S. cerevisiae when using carotenogenic genes from X. dendrorhous or E. uredovora.…”

Section: Resultsmentioning

confidence: 90%

“…In summary, we have been able to construct S. cerevisiae strains that produce various amounts of carotenoids by integration and overexpression of carotenogenic genes from X. dendrorhous. We succeeded in the construction of a strain producing 5.9 mg ␤-carotene/g (dw), which is 57-fold more than previously reported for heterologous ␤-carotene production in S. cerevisiae (41). This was achieved by overexpression of the catalytic domain of HMG1 from S. cerevisiae and additional overexpression of the crtI gene from X. dendrorhous in carotenoid-producing S. cerevisiae cells transformed with carotenogenic genes from X. dendrorhous.…”

Section: Resultsmentioning

confidence: 99%

“…In an initial effort to produce ␤-carotene heterologously in S. cerevisiae, the carotenogenic genes crtE, crtB, crtI, and crtY from the bacterium Erwinia uredovora were introduced and overexpressed from episomal vectors with different yeast-derived promoters and terminators for each gene. This resulted in quite low ␤-carotene production levels of 103 g/g (dry weight [dw]) (41). Overexpression of the same genes in the food yeast Candida utilis resulted in higher ␤-carotene production levels of 400 g/g (dw) (18).…”

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High-Level Production of Beta-Carotene in Saccharomyces cerevisiae by Successive Transformation with Carotenogenic Genes from Xanthophyllomyces dendrorhous

Verwaal

Wang

Meijnen

et al. 2007

Appl Environ Microbiol

324

303

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To determine whether Saccharomyces cerevisiae can serve as a host for efficient carotenoid and especially ␤-carotene production, carotenogenic genes from the carotenoid-producing yeast Xanthophyllomyces dendrorhous were introduced and overexpressed in S. cerevisiae. Because overexpression of these genes from an episomal expression vector resulted in unstable strains, the genes were integrated into genomic DNA to yield stable, carotenoid-producing S. cerevisiae cells. Furthermore, carotenoid production levels were higher in strains containing integrated carotenogenic genes. Overexpression of crtYB (which encodes a bifunctional phytoene synthase and lycopene cyclase) and crtI (phytoene desaturase) from X. dendrorhous was sufficient to enable carotenoid production. Carotenoid production levels were increased by additional overexpression of a homologous geranylgeranyl diphosphate (GGPP) synthase from S. cerevisiae that is encoded by BTS1. Combined overexpression of crtE (heterologous GGPP synthase) from X. dendrorhous with crtYB and crtI and introduction of an additional copy of a truncated 3-hydroxy-3-methylglutaryl-coenzyme A reductase gene (tHMG1) into carotenoid-producing cells resulted in a successive increase in carotenoid production levels. The strains mentioned produced high levels of intermediates of the carotenogenic pathway and comparable low levels of the preferred end product ␤-carotene, as determined by high-performance liquid chromatography. We finally succeeded in constructing an S. cerevisiae strain capable of producing high levels of ␤-carotene, up to 5.9 mg/g (dry weight), which was accomplished by the introduction of an additional copy of crtI and tHMG1 into carotenoid-producing yeast cells. This transformant is promising for further development toward the biotechnological production of ␤-carotene by S. cerevisiae.

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Section: Resultsmentioning

confidence: 90%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

High-Level Production of Beta-Carotene in Saccharomyces cerevisiae by Successive Transformation with Carotenogenic Genes from Xanthophyllomyces dendrorhous

Verwaal

Wang

Meijnen

et al. 2007

Appl Environ Microbiol

324

303

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“…Until now, most microbial production strategies have mostly used Escherichia coli as a host strain (1, 2, 10, 13, 24, 26, 30, 33-35, 43, 45, 48, 56-58, 64). However, a few studies using yeasts have also been reported (36,49,62). In early studies (46) with E. coli as a host strain, lycopene levels of up to about 0.5 mg/g (dry weight) of cells have been reported, whereas in Candida utilis, after extensive reengineering of the ergosterol pathway, up to 7.8 mg of lycopene/g (dry weight) has been achieved (49).…”

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confidence: 99%

High-Level Production of the Industrial Product Lycopene by the Photosynthetic Bacterium Rhodospirillum rubrum

Wang

Grammel

Abou-Aisha

et al. 2012

Appl Environ Microbiol

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bThe biosynthesis of the major carotenoid spirilloxanthin by the purple nonsulfur bacterium Rhodospirillum rubrum is thought to occur via a linear pathway proceeding through phytoene and, later, lycopene as intermediates. This assumption is based solely on early chemical evidence (B. H. Davies, Biochem. J. 116:93-99, 1970). In most purple bacteria, the desaturation of phytoene, catalyzed by the enzyme phytoene desaturase (CrtI), leads to neurosporene, involving only three dehydrogenation steps and not four as in the case of lycopene. We show here that the chromosomal insertion of a kanamycin resistance cassette into the crtCcrtD region of the partial carotenoid gene cluster, whose gene products are responsible for the downstream processing of lycopene, leads to the accumulation of the latter as the major carotenoid. We provide spectroscopic and biochemical evidence that in vivo, lycopene is incorporated into the light-harvesting complex 1 as efficiently as the methoxylated carotenoids spirilloxanthin (in the wild type) and 3,4,3=,4=-tetrahydrospirilloxanthin (in a crtD mutant), both under semiaerobic, chemoheterotrophic, and photosynthetic, anaerobic conditions. Quantitative growth experiments conducted in dark, semiaerobic conditions, using a growth medium for high cell density and high intracellular membrane levels, which are suitable for the conventional industrial production in the absence of light, yielded lycopene at up to 2 mg/g (dry weight) of cells or up to 15 mg/liter of culture. These values are comparable to those of many previously described Escherichia coli strains engineered for lycopene production. This study provides the first genetic proof that the R. rubrum CrtI produces lycopene exclusively as an end product.

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“…[57] Therefore, ERG9 was disrupted to switch the flux from ergosterol to lycopene, the target terpenoid. [58] In addition, crtE gene or HMG1 gene encoding HMGR was overexpressed, enhancing lycopene production.…”

Section: Increasing Upstream Fluxmentioning

confidence: 99%

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

Park

Yang

et al. 2017

Adv. Biosys.

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Natural products have been attracting much interest around the world for their diverse applications, especially in drug and food industries. Plants have been a major source of many different natural products. However, plants are affected by weather and environmental conditions and their successful extraction is rather limited. Chemical synthesis is inefficient due to the complexity of their chemical structures involving enantioselectivity and regioselectivity. For these reasons, an alternative means of overproducing valuable natural products using microorganisms has emerged. In recent years, various metabolic engineering strategies have been developed for the production of natural products by microorganisms. Here, the strategies taken to produce natural products are reviewed. For convenience, natural products are classified into four main categories: terpenoids, phenylpropanoids, polyketides, and alkaloids. For each product category, the strategies for establishing and rewiring the metabolic network for heterologous natural product biosynthesis, systems approaches undertaken to optimize production hosts, and the strategies for fermentation optimization are reviewed. Taken together, metabolic engineering has enabled microorganisms to serve as a prominent platform for natural compounds production. This article examines both the conventional and novel strategies of metabolic engineering, providing general strategies for complex natural compound production through the development of robust microbial‐cell factories.

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Metabolic Engineering for Production ofβ-Carotene and Lycopene inSaccharomyces cerevisiae

Cited by 144 publications

References 9 publications

High-Level Production of Beta-Carotene in Saccharomyces cerevisiae by Successive Transformation with Carotenogenic Genes from Xanthophyllomyces dendrorhous

High-Level Production of Beta-Carotene in Saccharomyces cerevisiae by Successive Transformation with Carotenogenic Genes from Xanthophyllomyces dendrorhous

High-Level Production of the Industrial Product Lycopene by the Photosynthetic Bacterium Rhodospirillum rubrum

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

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