Cloning and characterization of the astaxanthin biosynthetic gene encoding phytoene desaturase ofXanthophyllomyces dendrorhous

Verdoes, J.C.; Misawa, Norihiko; Ooyen, A.J.J. van

doi:10.1002/(sici)1097-0290(19990620)63:6<750::aid-bit13>3.0.co;2-7

Cited by 81 publications

(21 citation statements)

References 29 publications

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“…A bicyclic carotenogenic pathway in P. rhodozyma has been well documented [20–22]. A proposed scheme for astaxanthin synthesis in P. rhodozyma based on the present and previous studies is presented in Figure 5.…”

Section: Resultsmentioning

confidence: 70%

“…Synthesis starts from acetyl coenzyme A, then enters a terpenoid synthesis pathway [23]. Conversion of isoprenoid precursors into β-carotene occurs through four sequential enzymatic steps, catalyzed by GGPP synthase (encoded by crtE ), phytoene synthase (encoded by crtYB ), phytoene desaturase (encoded by crtI ), and lycopene cyclase (also encoded by crtYB ) [21, 22]. A monocyclic carotenoid biosynthesis pathway (also shown in Figure 5) was proposed [24].…”

Section: Resultsmentioning

confidence: 99%

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Overexpression of a bifunctional enzyme, CrtS, enhances astaxanthin synthesis through two pathways in Phaffia rhodozyma

Chi

Ren

et al. 2015

Microb Cell Fact

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BackgroundA moderate-temperature, astaxanthin-overproducing mutant strain (termed MK19) of Phaffia rhodozyma was generated in our laboratory. The intracellular astaxanthin content of MK19 was 17-fold higher than that of wild-type. The TLC profile of MK19 showed a band for an unknown carotenoid pigment between those of β-carotene and astaxanthin. In the present study, we attempted to identify the unknown pigment and to enhance astaxanthin synthesis in MK19 by overexpression of the crtS gene that encodes astaxanthin synthase (CrtS).ResultsA crtS-overexpressing strain was constructed without antibiotic marker. A recombinant plasmid with lower copy numbers was shown to be stable in MK19. In the positive recombinant strain (termed CSR19), maximal astaxanthin yield was 33.5% higher than MK19, and the proportion of astaxanthin as a percentage of total carotenoids was 84%. The unknown carotenoid was identified as 3-hydroxy-3′,4′-didehydro-β,Ψ-carotene-4-one (HDCO) by HPLC, mass spectrometry, and NMR spectroscopy. CrtS was found to be a bifunctional enzyme that helped convert HDCO to astaxanthin. Enhancement of crtS transcriptional level increased transcription levels of related genes (crtE, crtYB, crtI) in the astaxanthin synthesis pathway. A scheme of carotenoid biosynthesis in P. rhodozyma involving alternative bicyclic and monocyclic pathways is proposed.ConclusionsCrtS overexpression leads to up-regulation of synthesis-related genes and increased astaxanthin production. The transformant CSR19 is a stable, secure strain suitable for feed additive production. The present findings help clarify the regulatory mechanisms that underlie metabolic fluxes in P. rhodozyma carotenoid biosynthesis pathways.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-015-0279-4) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 70%

Section: Resultsmentioning

confidence: 99%

Overexpression of a bifunctional enzyme, CrtS, enhances astaxanthin synthesis through two pathways in Phaffia rhodozyma

Chi

Ren

et al. 2015

Microb Cell Fact

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“…A more general role applicable to both photosynthetic and non-photosynthetic cell, is protection from photodynamic action [1]. Genes controlling the synthesis of these pigments have been studied in several organisms such as Erwinia species [2, 3, 4], Mycobacterium aurum A + [5, 6], Arabidopsis [7, 8], Xantophyllomyces dendrorhous [9] and Brevibacterium linens [10]. …”

Section: Introductionmentioning

confidence: 99%

Response of Escherichia Coli Containing Mycobacterial Carotene Genes to UV Radiation

Houssaini-Iraqui

Khamlichi

Yamani

et al. 2001

BioMed Research International

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The plasmid pC5, which encodes biogenesis of lycopene in Mycobacterium aurum A + , was partially digested by restriction endonucleases and generated fragments were cloned. After transformation of Escherichia coli (colorless bacteria) with the plasmids so constructed, seven orange clones were detected and found to carry the same recombinant plasmid (pC51). E. coli cells containing this plasmid synthesize neurosporene and lycopene, and were more resistant to ultraviolet irradiation than nonpigmented strain.

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“…Phytoene is the first carotenoid synthesized in the pathway, produced from GGPP by the bifunctional enzyme phytoene-β-carotene synthase (PBS, encoded by the crtYB gene) [11]. Then, phytoene is transformed to lycopene by phytoene desaturase (encoded by the crtI gene) [12]. Subsequently, both ends of lycopene are cyclized by PBS to produce β-carotene [11].…”

Section: Introductionmentioning

confidence: 99%

Regulation of carotenogenesis in the red yeast Xanthophyllomyces dendrorhous: the role of the transcriptional co-repressor complex Cyc8–Tup1 involved in catabolic repression

et al. 2016

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BackgroundThe yeast Xanthophyllomyces dendrorhous produces carotenoids of commercial interest, including astaxanthin and β-carotene. Although carotenogenesis in this yeast and the expression profiles of the genes controlling this pathway are known, the mechanisms regulating this process remain poorly understood. Several studies have demonstrated that glucose represses carotenogenesis in X. dendrorhous, suggesting that this pathway could be regulated by catabolic repression. Catabolic repression is a highly conserved regulatory mechanism in eukaryotes and has been widely studied in Saccharomyces cerevisiae. Glucose-dependent repression is mainly observed at the transcriptional level and depends on the DNA-binding regulator Mig1, which recruits the co-repressor complex Cyc8–Tup1, which then represses the expression of target genes. In this work, we studied the regulation of carotenogenesis by catabolic repression in X. dendrorhous, focusing on the role of the co-repressor complex Cyc8–Tup1.ResultsThe X. dendrorhous CYC8 and TUP1 genes were identified, and their functions were demonstrated by heterologous complementation in S. cerevisiae. In addition, cyc8 − and tup1 − mutant strains of X. dendrorhous were obtained, and both mutations were shown to prevent the glucose-dependent repression of carotenogenesis in X. dendrorhous, increasing the carotenoid production in both mutant strains. Furthermore, the effects of glucose on the transcript levels of genes involved in carotenogenesis differed between the mutant strains and wild-type X. dendrorhous, particularly for genes involved in the synthesis of carotenoid precursors, such as HMGR, idi and FPS. Additionally, transcriptomic analyses showed that cyc8 − and tup1 − mutations affected the expression of over 250 genes in X. dendrorhous. ConclusionsThe CYC8 and TUP1 genes are functional in X. dendrorhous, and their gene products are involved in catabolic repression and carotenogenesis regulation. This study presents the first report involving the participation of Cyc8 and Tup1 in carotenogenesis regulation in yeast.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0597-1) contains supplementary material, which is available to authorized users.

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Cloning and characterization of the astaxanthin biosynthetic gene encoding phytoene desaturase ofXanthophyllomyces dendrorhous

Cited by 81 publications

References 29 publications

Overexpression of a bifunctional enzyme, CrtS, enhances astaxanthin synthesis through two pathways in Phaffia rhodozyma

Overexpression of a bifunctional enzyme, CrtS, enhances astaxanthin synthesis through two pathways in Phaffia rhodozyma

Response of Escherichia Coli Containing Mycobacterial Carotene Genes to UV Radiation

Regulation of carotenogenesis in the red yeast Xanthophyllomyces dendrorhous: the role of the transcriptional co-repressor complex Cyc8–Tup1 involved in catabolic repression

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