[go: up one dir, main page]

WO2019016383A1 - Nouvelle ferrédoxine - Google Patents

Nouvelle ferrédoxine Download PDF

Info

Publication number
WO2019016383A1
WO2019016383A1 PCT/EP2018/069802 EP2018069802W WO2019016383A1 WO 2019016383 A1 WO2019016383 A1 WO 2019016383A1 EP 2018069802 W EP2018069802 W EP 2018069802W WO 2019016383 A1 WO2019016383 A1 WO 2019016383A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
rhodoxanthin
seq
carotene
beta
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2018/069802
Other languages
English (en)
Inventor
Bastien Chevreux
John Royer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DSM IP Assets BV
Original Assignee
DSM IP Assets BV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by DSM IP Assets BV filed Critical DSM IP Assets BV
Publication of WO2019016383A1 publication Critical patent/WO2019016383A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0095—Oxidoreductases (1.) acting on iron-sulfur proteins as donor (1.18)

Definitions

  • the present invention is related to novel protein involved in transfer of electrons, in particular in an enzymatic conversion of carotenoids into particular retro- carotenoids, more particularly in a conversion of beta-carotene into rhodoxanthin via the catalytic action of rhodoxanthin-producing hydroxylases (hereinafter also referred to as BHYRs).
  • BHYRs rhodoxanthin-producing hydroxylases
  • the invention also features polynucleotides and the corresponding polypeptides comprising the full-length sequences of the novel gene/polypeptide and fragments thereof, in particular functional equivalents of said gene/polypeptide.
  • the invention further relates to genetically engineered carotene- producing fungal host cells and their use for biotechnological production of rhodoxanthin.
  • Examples of such compounds acting as donor and/or acceptor of electrons include but are not limited to co-factors and/or proteins selected from the group consisting of NAD(H), NADP(H), ferredoxin, ferredoxin oxidoreductase, flavodoxin, flavodoxin oxidoreductase, putaredoxin, putaredoxin reductase,
  • Retro-carotenoids are carotenoids with a shift of one position of the single and double bonds of their respective conjugated polyene systems.
  • Rhodoxanthin an example of a retro-carotenoid, and which is found in nature in e.g. arils, berries, leaves or flowers of the poisonous yew (Taxus), Aloe or honeysuckle (Lonicera sp.), is widely used as a coloring material for foodstuffs and beverages as well as pharmaceutical and cosmetic preparations, imparting to them a yellow to red coloration.
  • yew poisonous yew
  • Aloe or honeysuckle Loa suckle
  • As a food additive it is used under the E number E161f as a food coloring.
  • E161f As a food coloring.
  • rhodoxanthin and other retro-carotenoids which can be used as e.g. coloring material in the food & beverage, pharmaceutical and cosmetic industry in order to replace the chemically produced (synthetic) rhodoxanthin.
  • a rhodoxanthin-producing beta-carotene hydroxylase (herein referred as BHYR) has been identified from the red berries of Lonicera (SEQ ID NO:4) with the aim to biotechnologically generate rhodoxanthin in the respective host cell, such as e.g. fungi, in particular strains of Yarrowia, more preferably Y. lipolytica,
  • BHYR beta-carotene hydroxylase
  • a suitable host cell such as e.g. a carotenoid-producing fungal cell including yeast
  • the present invention is directed to a polynucleotide having electron transfer activity, i.e. ferredoxin activity, which is selected from a polypeptide with at least 59%, such as e.g. at least 60, 65, 70, 75, 80, 90, 92, 95, 98, 99% or up to 100% identity to SEQ ID NO:2, which might be encoded by a polynucleotide including, but not limited to SEQ ID NO:1 or SEQ ID NO:1 1 , in particular a recombinant nucleic acid molecule.
  • the present invention is directed to the use of the novel protein having ferredoxin activity in a biotechnological production of rhodoxanthin in a suitable host cell, in particular a carotenoid-producing fungal host including yeast, wherein said host cell furthermore is expressing a gene encoding a polypeptide having beta-carotene hydroxylase activity and which is selected from a polypeptide with at least 63%, such as e.g. at least 65, 70, 75, 80, 85, 90, 95, 97, 98, 99 or even 100% identity to SEQ ID NO:4, which might be encoded by a polynucleotide including, but not limited to SEQ ID NO:3, in particular a
  • the present invention is directed to the use of the novel protein having ferredoxin reductase activity in a biotechnological production of rhodoxanthin in a suitable host cell, in particular a carotenoid-producing fungal host including yeast, wherein said host cell furthermore is expressing a gene encoding a polypeptide having beta-carotene hydroxylase activity and which is selected from a polypeptide with at least 78%, such as e.g. 80, 85, 90, 95, 97, 98, 99 or even 100% identity to SEQ ID NO:6, which might be encoded by a
  • polynucleotide including, but not limited to SEQ ID NO:5, in particular a
  • the protein having ferredoxin activity is obtainable from Aloe as shown in SEQ ID NO:2, in particular encoded by a polynucleotide according to SEQ ID NO:1 or SEQ ID NO:1 1 .
  • said protein with at least 59 % identity to SEQ ID NO:2 is hereinafter defined as "Aloe ferredoxin” or "Aloe FD”.
  • the enzyme according to SEQ ID NO:2 shows an amino acid identity of only 58.3 % to the known ferredoxin protein from Arabidopsis thaliana (uniprotKB/Swiss-Prot O04090).
  • the present invention is related to a process for conversion of beta-carotene into rhodoxanthin, said conversion being catalyzed by a protein having at least 63%, such as e.g.65, 70, 75, 80, 85, 90, 95, 97, 98, 99% or up to 100% identity to SEQ ID NO:4, which might be encoded by a polynucleotide including, but not limited to SEQ ID NO:3, in particular a recombinant nucleic acid molecule, said conversion being performed in the presence of the Aloe FD as described herein.
  • expressing a polypeptide having beta-carotene hydroxylase activity with at least 63 %, such as e.g. 65, 70, 75, 80, 85, 90, 95, 97, 98, 99% or up to 100% identity to SEQ ID NO:4 together with a polypeptide having ferredoxin activity with at least 59 % identity to SEQ ID NO:2 can be selected from any fungal host suitable for carotenoid production, including carotenoid-producing yeast cell such as e.g. Yarrowia.
  • a carotenoid-producing host cell is a host cell wherein the respective polypeptides are expressed and active in vivo leading to production of carotenoids.
  • the genes and methods to generate carotenoid-producing host cells are known in the art. Depending on the carotenoid to be produced, different genes might be involved.
  • a rhodoxanthin-producing host cell is capable of expressing a polypeptide having beta-carotene hydroxylase activity with at least 63 %, such as e.g. 65, 70, 75, 80, 85, 90, 95, 97, 98, 99% or up to 100% identity to SEQ ID NO:4.
  • fungi including hosts of the known culture collections as published by the World Federation for Culture Collections (WFCC).
  • WFCC World Federation for Culture Collections
  • a fungal cell including yeast, is selected from the group consisting of Saccharomyces, Aspergillus, Pichia, Hansenula, Phycomyces, Mucor,
  • Rhodotorula Rhodotorula, Sporobolomyces, Xanthophyllomyces, Phaffia, Blakeslea, and Yarrowia, preferably selected from the group consisting of Saccharomyces cerevisiae, Aspergillus niger, Pichia pastoris, Hansenula polymorpha, Phycomyces blakesleanus, Blakeslea trispora and Yarrowia lipolytica, In particularly preferred is expression in Yarrowia, most preferably expression in Yarrowia lipolytica.
  • microorganisms also include synonyms or basonyms of such species having the same physiological properties, as defined by the International Code of Nomenclature of Prokaryotes or the International Code of Nomenclature for algae, fungi, and plants (Melbourne Code).
  • % identity refers to the comparison of two amino acid sequences using a sequence analysis program as for instance Blast or Clustal Omega. Secondary structure prediction can be done by at least use of the Prime software from Schrodinger or by on-line software tools such as JPred.
  • % identical refers to the percent of the amino acids of the subject amino acid sequence that have been matched to identical amino acids in the compared amino acid sequence. If both amino acid sequences which are compared do not differ in any of their amino acids, they are identical or have 100% identity.
  • the present invention is directed to a novel ferredoxin as defined herein involved in electron transfer in the enzymatic conversion of carotenoids into rhodoxanthin, said bioconversion being catalyzed by a beta-carotene
  • BHYR hydroxylating enzyme
  • the BHYR gene might be truncated and/or codon-optimized for expression in the fungal host cell, such as e.g. Yarrowia (see SEQ ID NO:5).
  • the rhodoxanthin-producing host cell in particular a
  • carotenoid-producing fungal host cell including yeast, preferably Yarrowia, furthermore comprises a protein having ferredoxin-NADP reductase activity, in particular wherein the gene encoding said polypeptide being originating from Aloe, preferably with at least 82%, such as e.g. 85, 90, 92, 95, 97, 99 or event up to 100% identity to SEQ ID NO:8, which might be encoded by a polynucleotide including, but not limited to SEQ ID NO:7 or 12.
  • the rhodoxanthin-producing host cell in particular a carotenoid-producing fungal host cell including yeast, preferably Yarrowia, furthermore comprises a protein having chloroplast processing enzyme (CPE) activity, such as, e.g. a gene encoding stromal processing peptidase (UniProtKB Q40983) from Pisum sativum, preferably with at least 80%, such as 85, 90, 92, 95, 97, 99 or even 100% identity to SEQ ID NO:10, which might be encoded by a polynucleotide including, but not limited to SEQ ID NO:9.
  • CPE chloroplast processing enzyme
  • SEQ ID NO:4 a polynucleotide expressing a polypeptide having ferredoxin reductase activity with at least 82%, such as e.g. 85, 90, 92, 95, 97, 99 or event up to 100% identity to SEQ ID NO:8, and (d) a polynucleotide expressing a polypeptide having CPE activity with at least 80 %, such as 85, 90, 92, 95, 97, 99 or even 100% identity SEQ ID NO:10.
  • ferredoxin activity encompasses activity of all three ferredoxin isoforms, i.e. as ferredoxin 1 , 2 and/or 3, including leaf and
  • the polynucleotides expressing BHYR, Aloe FD, Aloe ferredoxin reductase or CPE as defined herein might be optimized for expression in the respective host cell.
  • the skilled person knows how to generate such modified polynucleotides.
  • the polynucleotides as defined herein also encompass such host-optimized nucleic acid molecules as long as they still express the polypeptide with the respective activities as defined herein.
  • the novel enzyme according to the present invention also encompasses enzymes carrying amino acid substitution(s) which do not alter enzyme activity, i.e. which show the same properties with respect to the wild-type enzyme and catalyze at least one of the above mentioned oxidation reduction reactions.
  • Such mutations are also called "silent mutations", which do not alter the (enzymatic) activity of the enzyme as described herein.
  • a nucleic acid molecule according to the invention may comprise only a portion or a fragment of the nucleic acid sequence provided by the present invention, such as for instance the sequence shown in SEQ ID NO:1 or SEQ ID NO:1 1 for example a fragment which may be used as a probe or primer or a fragment encoding a portion of a protein according to the invention.
  • the nucleotide sequence determined from the cloning of the FD gene allows for the generation of probes and primers designed for use in identifying and/or cloning other FD family members, as well as FD homologues from other species.
  • the probe/primer typically comprises substantially purified oligonucleotides which typically
  • nucleotide sequence that hybridizes preferably under highly stringent conditions to at least about 12 or 15, preferably about 18 or 20, more preferably about 22 or 25, even more preferably about 30, 35, 40, 45, 50, 55, 60, 65, or 75 or more consecutive nucleotides of a nucleotide sequence shown in SEQ ID NO:1 or SEQ ID NO:1 1 or a fragment or derivative thereof.
  • a preferred, non-limiting example of such hybridization conditions are hybridization in 6x sodium chloride/sodium citrate (SSC) at about 45°C, followed by one or more washes in 1 x SSC, 0.1 % SDS at 50°C, preferably at 55°C, more preferably at 60°C and even more preferably at 65°C.
  • Highly stringent conditions include, for example, 2 h to 4 days incubation at 42°C using a digoxigenin (DIG)-labeled DNA probe (prepared by using a DIG labeling system; Roche Diagnostics GmbH, 68298 Mannheim, Germany) in a solution such as DigEasyHyb solution (Roche Diagnostics GmbH) with or without 100 g/ml salmon sperm DNA, or a solution comprising 50% formamide, 5x SSC (150 mM NaCI, 15 mM trisodium citrate), 0.02% sodium dodecyl sulfate, 0.1 % N- lauroylsarcosine, and 2% blocking reagent (Roche Diagnostics GmbH), followed by washing the filters twice for 5 to 15 minutes in 2x SSC and 0.1 % SDS at room temperature and then washing twice for 15-30 minutes in 0.5x SSC and 0.1 % SDS or 0.1 x SSC and 0.1 % SDS at 65-68°C.
  • DIG digoxigen
  • the host cell such as e.g. a carotenoid-producing fungal cell, which is able to express the BHYR, FD, Aloe ferredoxin reductase with or without CPE according to the present invention may be cultured in an aqueous medium supplemented with appropriate nutrients under aerobic or anaerobic conditions and as known by the skilled person for the different host cells.
  • such cultivation is in the presence of proteins and/or co-factors involved in transfer of electrons, as defined herein.
  • the cultivation/growth of the host cell may be conducted in batch, fed- batch, semi-continuous or continuous mode.
  • production of retro-carotenoids such as e.g. rhodoxanthin is performed in a fed-batch process using corn oil as carbon source.
  • a process according to the present invention is directed to production of rhodoxanthin from beta-carotene, comprising the steps of:
  • ferredoxin as defined herein from Aloe in a suitable carotenoid- producing (fungal) host cell
  • said protein being preferably a polypeptide according to SEQ ID NO:2 or a polypeptide with at least 59%, such as e.g. at least 60, 65, 70, 75, 80, 90, 92, 95, 98, 99% or up to 100% identity to SEQ ID NO:2;
  • a polypeptide having activity towards conversion of beta-carotene into rhodoxanthin such as BHYR as defined herein, preferably a BHYR with at least 63 %, such as e.g. 65, 70, 75, 80, 85, 90, 95, 97, 98, 99 or even 100% identity to SEQ ID NO:4,
  • ferredoxin reductase as defined herein from Aloe, said protein being preferably a polypeptide according to SEQ ID NO:8 or a polypeptide with at least 82%, preferably at least 85, 90, 92, 95, 97, 99 or event up to 100% identity to SEQ ID NO:8,
  • CPE as defined herein from Pisum sativum, said protein being preferably a polypeptide according to SEQ ID NO:10 or a
  • polypeptide with at least 80%, preferably at least 85, 90, 92, 95, 97, 99 or even 100% identity to SEQ ID NO:10,
  • the term "specific activity” or "activity” with regards to enzymes means its catalytic activity, i.e. its ability to catalyze formation of a product from a given substrate.
  • the specific activity defines the amount of substrate consumed and/or product produced in a given time period and per defined amount of protein at a defined temperature.
  • specific activity is expressed in ⁇ substrate consumed or product formed per min per mg of protein.
  • An enzyme is active, if it performs its catalytic activity in vivo, i.e.
  • FD activity is defined as the capability to facilitate the electron transfer in the oxidation of beta-carotene into rhodoxanthin.
  • FD activity is also defined as the capability to facilitate the electron transfer in the production of other plant metabolites, carotenoid, retrocarotenoid such as eschscholtzanthin.
  • Rhodoxanthin as used herein includes any chemical form of rhodoxanthin found in aqueous solutions, including all isoforms. In particular, it includes a mixture of cis EE-, EZ and ZZ-rhodoxanthin.
  • FIG. 1 HPLC chromatogram (left side) and UV spectra for the individual peaks (right side) of Yarrowia lipolytica extracts.
  • A is compared to Y. lipolytica transformed with CarB and CarRP from Mucor circinellioides, BHYr from Lonicera sp, CPE from Pisum sativum, ferredoxin (Fd2) and ferredoxin reductase (FNR) from Aloe sp.
  • Peak 1 beta-cryptoxanthin
  • peak 2 peak 2:
  • rhodoxanthin cis ee isomer peak 5: zeaxanthin.
  • Absorbance at 494 nm is shown on the y-axis, time in minutes is shown on the x-axis.
  • a transcriptomics approach was used to identify genes that were highly expressed in rhodoxanthin-producing flowers of Pink Blush Aloe.
  • a gene (Fd2 SEQ ID NO:1 1 ) with homology to ferredoxins was found to be highly expressed.
  • Yarrowia codon optimized version of the gene (Fd2 SEQ ID NO:1 ) with restriction sites for cloning into Yarrowia expression vectors driven by the Tefl promoter was synthesized (Genscript) and cloned into a nourseothricin expression vector to generate plasmid pMB8059
  • Ferredoxin homologues can be identified from the publicly available protein and nucleic acid databases.
  • the Aloe FD DNA sequence according to SEQ ID NO:1 1 can be used as probe in a Southern blot to identify homologous FD clones from other organisms, in particular from organisms which are able to synthesize carotenoids, in particular rhodoxanthin or other retrocarotenoids from beta-carotene.
  • a standard protocol for Southern blot hybridization is described in "Molecular cloning: A lab manual. Sambrook, J, E. F. Fritsch, T. Maniatis 1989" Identified clones are further tested together for their activity on electron transfer in the presence of a rhodoxanthin forming protein (BHYR) as defined in SEQ ID NO:4 or SEQ ID NO:6.
  • BHYR rhodoxanthin forming protein
  • Example3 Expression of FD and production of rhodoxanthin in a yeast system
  • Rhodoxanthin production in Yarrowia lipolytica was enabled by introducing the full- length Aloe FD gene into a beta-carotene producing Yarrowia host strain, in addition comprising DNA expressing BHYR, full-length Aloe ferredoxin reductase along with DNA expressing the chloroplast processing enzyme from Pisum sativum designed to cleave the chloroplast targeting signal from the ferredoxin and ferredoxin reductase.
  • the BHYR gene of Lonicera (SEQ ID NO:3) was truncated to remove the presumed chloroplast targeting signal, and modified by codon optimization for expression in Yarrowia to result in SEQ ID NO:5.
  • An Nhel site was incorporated into the 5'-region, and an Mlul site was incorporated into the 3'-region to allow subcloning into the Yarrowia expression vector MB6157 to generate plasmid pMB7918, where expression of BHYR is under the control of the Yarrowia Tef promoter.
  • MB7918 was transformed into the beta-carotene producing strain ML2461 which contains Mucor circinellioides CarB (phytoene dehydrogenase; CAB40843.1 ) and M. circinellioides CarRP (lycopene cyclase/phytoene synthase; CAB60272.1 ) to generate strain ML17461 .
  • ML17461 was grown on YPoil (yeast extract (10 g/l), peptone (20 g/l), tryptophan (0.15 g/l) and corn oil 2%) medium and samples were extracted and analyzed by normal phase HPLC. Small amounts of beta-cryptoxanthin, zeaxanthin, and tentative rhodoxanthin EZ were produced (Fig.l A).
  • DNA for Aloe ferredoxin, Aloe ferredoxin reductase and P. sativum CPE genes were Yarrowia codon-optimized (see SEQ ID NOs:1 , 7, 9) and synthesized with an Nhel site at the 5'-end and an Mlul site at the 3'-end to allow cloning into Yarrowia expression vectors to generate pMB8059, pMB8056, and pMB7896. Expression of FD and FNR was with the TEF promoter, while expression of CPE was with the Alk1 promoter of Yarrowia.
  • Strain ML 17769 which contains the Mucor CarB and CarRP genes for beta carotene production, the P. sativum chloroplast targeting enzyme (CPE), the Aloe ferredoxin and ferredoxin reductase genes, and the truncated BHYR gene was grown on YPoil (yeast extract (10 g/l), peptone (20 g/l), tryptophan (0.15 g/l) and corn oil 2%) medium and samples were extracted and analyzed by normal phase HPLC. ML17769 produced significantly higher levels of rhodoxanthin than strain ML17461 containing CarB, CarRP and BHYR, alone (Fig.1 B).
  • CPE P. sativum chloroplast targeting enzyme
  • Aloe ferredoxin and ferredoxin reductase genes the truncated BHYR gene was grown on YPoil (yeast extract (10 g/l), peptone (20 g/l), tryptophan (0.15 g/

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne une nouvelle protéine impliquée dans le transfert d'électrons, en particulier dans une conversion enzymatique de caroténoïdes en rétro-caroténoïdes particuliers, plus particulièrement dans une conversion de bêta-carotène en rhodoxanthine par l'intermédiaire de l'action catalytique d'hydroxylases produisant de la rhodoxanthine (également désignées ci-après par BHYR). L'invention concerne également des polynucléotides et les polypeptides correspondants comprenant les séquences pleines longueurs du nouveau gène/polypeptide et des fragments de celui-ci, en particulier des équivalents fonctionnels dudit gène/polypeptide. L'invention concerne en outre des cellules hôtes fongiques génétiquement modifiées productrices de carotène et leur utilisation pour la production biotechnologique de rhodoxanthine.
PCT/EP2018/069802 2017-07-21 2018-07-20 Nouvelle ferrédoxine Ceased WO2019016383A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201762535631P 2017-07-21 2017-07-21
US62/535,631 2017-07-21

Publications (1)

Publication Number Publication Date
WO2019016383A1 true WO2019016383A1 (fr) 2019-01-24

Family

ID=62986114

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2018/069802 Ceased WO2019016383A1 (fr) 2017-07-21 2018-07-20 Nouvelle ferrédoxine

Country Status (1)

Country Link
WO (1) WO2019016383A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025207999A3 (fr) * 2024-03-29 2025-11-20 Inscripta, Inc. Compositions et procédés pour améliorer la production de carotenoïdes

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6117666A (en) * 1995-08-01 2000-09-12 Arch Development Corporation Plastid proteolytic processing enzyme that cleaves precursor polypeptides
WO2006068946A2 (fr) * 2004-12-20 2006-06-29 Boyce Thompson Institute For Plant Research Augmentation de la teneur en beta-carotene dans des plantes
WO2009009142A2 (fr) * 2007-07-10 2009-01-15 Monsanto Technology, Llc Plantes transgéniques à caractéristiques agronomiques améliorées

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6117666A (en) * 1995-08-01 2000-09-12 Arch Development Corporation Plastid proteolytic processing enzyme that cleaves precursor polypeptides
WO2006068946A2 (fr) * 2004-12-20 2006-06-29 Boyce Thompson Institute For Plant Research Augmentation de la teneur en beta-carotene dans des plantes
WO2009009142A2 (fr) * 2007-07-10 2009-01-15 Monsanto Technology, Llc Plantes transgéniques à caractéristiques agronomiques améliorées

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE UniProt [online] 30 November 2016 (2016-11-30), SUZUKI J.: ""Transcriptome Assembly of Anthurium amnicola.";", XP055517209, Database accession no. A0A1D1YVI3 *
MISAWA N ET AL: "Metabolic engineering for the production of carotenoids in non-carotenogenic bacteria and yeasts", JOURNAL OF BIOTECHNOLOGY, ELSEVIER, AMSTERDAM, NL, vol. 59, no. 3, 3 January 1998 (1998-01-03), pages 169 - 181, XP004113748, ISSN: 0168-1656, DOI: 10.1016/S0168-1656(97)00154-5 *
SAMBROOK, J, E. F. FRITSCH; T. MANIATIS: "Molecular cloning: A lab manual", 1989
ZHOU PINGPING ET AL: "Highly efficient biosynthesis of astaxanthin inSaccharomyces cerevisiaeby integration and tuning of algalcrtZandbkt", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER, DE, vol. 99, no. 20, 9 July 2015 (2015-07-09), pages 8419 - 8428, XP035547321, ISSN: 0175-7598, [retrieved on 20150709], DOI: 10.1007/S00253-015-6791-Y *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025207999A3 (fr) * 2024-03-29 2025-11-20 Inscripta, Inc. Compositions et procédés pour améliorer la production de carotenoïdes

Similar Documents

Publication Publication Date Title
JP7443656B2 (ja) レチニルエステルの生産
JP7513520B2 (ja) レチノールの生産
JP7472427B2 (ja) レチノイドの生合成
KR102202606B1 (ko) 바이오레티놀을 생산하는 미생물 및 이를 이용한 바이오레티놀의 생산방법
Sandmann Carotenoid biosynthesis in microorganisms and plants
CN111107833A (zh) 反式视黄醛的生产
KR19990088053A (ko) 이소프레노이드의향상된생산방법
EP3906305A1 (fr) Nouvelles acétyl-transférases
EP3999634A1 (fr) Nouvelles bêta-carotène oxydases
US20050124031A1 (en) Biosyntheticse genes of blakeslea trispora beta-carotene that code for lycopene cyclase/phytoene synthase (carrp) and phytoene dehydrogenase (carb)
AU2016294344A1 (en) Microorganisms with broadened light absorption capability and increased photosynthetic activity
JP7298812B2 (ja) レチニルエステルの生産
WO2019016383A1 (fr) Nouvelle ferrédoxine
WO2019016382A1 (fr) Nouvelle ferredoxine réductase
Li et al. Comparative transcriptome analysis revealed the improved β-carotene production in Sporidiobolus pararoseus yellow mutant MuY9
WO2019016384A1 (fr) Nouvelle enzyme
KR100745216B1 (ko) 신규한 베타-카로틴 케톨라제 및 베타-카로틴하이드록실라제 유전자
JP2026502492A (ja) Ｃａｒｒｐ酵素バリアント、並びにカロテノイド及びアポカロテノイドを産生する際のそれらの使用
EP3687486A1 (fr) Biosynthèse de rétinoïdes

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18743792

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18743792

Country of ref document: EP

Kind code of ref document: A1