EP1161542A1 - Elongase für mehrfach ungesättigte fettsäuren aus caenorhabditis elegans - Google Patents

Elongase für mehrfach ungesättigte fettsäuren aus caenorhabditis elegans

Info

Publication number
EP1161542A1
EP1161542A1 EP00911091A EP00911091A EP1161542A1 EP 1161542 A1 EP1161542 A1 EP 1161542A1 EP 00911091 A EP00911091 A EP 00911091A EP 00911091 A EP00911091 A EP 00911091A EP 1161542 A1 EP1161542 A1 EP 1161542A1
Authority
EP
European Patent Office
Prior art keywords
polypeptide
pufa
acid
polypeptide according
animal
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.)
Withdrawn
Application number
EP00911091A
Other languages
English (en)
French (fr)
Inventor
J.A. IACR - Long Ashton Research Station NAPIER
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.)
University of Bristol
Original Assignee
University of Bristol
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
Priority claimed from GBGB9906307.5A external-priority patent/GB9906307D0/en
Priority claimed from GB0003869A external-priority patent/GB0003869D0/en
Application filed by University of Bristol filed Critical University of Bristol
Publication of EP1161542A1 publication Critical patent/EP1161542A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to polyunsaturated fatty acid (PUFA) elongases. More specifically, the invention relates to a DNA sequence from C. elegans encoding a PUFA elongase.
  • PUFA polyunsaturated fatty acid
  • Unsaturated fatty acids are essential components required for normal cellular function, being involved in a diverse number of roles ranging from membrane fluidity to acting as signal molecules (Gill, I., Valivety, R. (1997). Trends Biotechnol. 15, 401-409; Broun, P., et al (1999) Ann. Rev. Nutr. 19, 197-216).
  • the class of fatty acids known as the polyunsaturated fatty acids (PUFAs) has attracted considerable interest as pharmaceutical and nutraceutical compounds (Broun supra; Horrobin, D. F. (1990) Reviews in Contemp Pharmacotherpy 1, 1-45).
  • PUFAs i.e. fatty acids of 18 carbons or more in length and containing two or more double bonds
  • This elongase is responsible for the addition of 2 carbon units to an 18 carbon PUFA, resulting in a 20 carbon fatty acid.
  • This reaction is the elongation of ⁇ -linolenic acid (GLA; 18:3 ⁇ 6,9 12 ) to di-homo- ⁇ -linolenic acid (DHGLA; 20:3 ⁇ 8,11 ' 14 ) in which the tri-unsaturated 18 carbon fatty acid is elongated by the addition of a two carbon unit to yield the tri-unsaturated 20 carbon fatty acid. Since there is considerable interest in the production of long chain PUFAs of more than 18 carbons in chain length, for example arachidonic acid and eicosapentanoic acid, the identification of this enzyme is of both academic and commercial interest.
  • an Arabidopsis gene (FAE1) has been shown to be required for the synthesis of very long chain monounsaturated fatty acids (such as erucic acid; 20:l ⁇ ⁇ ) (James, D. W. et al, (1995) Plant Cell 7, 309-319).
  • this enzyme does not recognize di- and tri-unsaturated 18 carbon fatty acids, for example, linoleic acid, 18:2 ⁇ 9 ' 12 or ⁇ -linolenic acid, 18:3 ⁇ 9 ' 2 ' 5 respectively, as substrates, and is therefore not involved in the synthesis of long chain PUFAs (Millar & Kunststoff (1997), Plant Journal 12, 121-131).
  • FIG. 1 A schematic diagram representing a generalized pathway for the product of PUFAs is shown in Figure 1.
  • Biochemical characterisation of mammalian elongation systems has indicated that a mammalian elongase consists of four subunits, made up of a condensing enzyme, a ⁇ -ketoreductase, a dehydrase and an enoyl reductase (reviewed in Cinti, D. L., et al (1992) Prog. Lipid Res. 31, 1-51).
  • the Arabidopsis FAEl gene product encodes a polypeptide of 56kDa, which shows very limited homology to condensing enzymes such as chalcone synthase and stillbene synthase (James, D. W. supra). Although FAEl is normally only expressed in seed tissues, ectopic expression in non-seed tissue (or heterologously in yeast) revealed that FAEl could direct the synthesis of erucic acid (Millar, A. A., Kunststoff, L. (1997) Plant J. 12, 121-131).
  • ELO1 Three fatty acid elongase activities have been characterised from the yeast S. cerevisiae. Again, this organism does not synthesis PUFAs, and therefore does not contain genes encoding a PUFA elongase.
  • One gene ELO1 was identified on the basis of a screen to isolate mutants defective in elongation of 14 carbon (i.e. medium) chain saturated fatty acids (Toke & Martin (1996) J Biol Chem 271, 18413-18422). Complementation of elol mutants restored viability, and the ELO1 gene product was shown to encode a polypeptide which was responsible for the specific elongation of 14:0 fatty acids to 16:0 fatty acids.
  • ELO2 and ELO3 Two related genes were also detected in the genome of S. cerevisiae, and their function determined by disruption. These two genes, subsequently named ELO2 and ELO3, were shown to be involved in the elongation of the very long chain saturated fatty acids found in sphingolipid molecules (Oh et al (1997), J. Biol Chem 272, 17376-17384). In particular, ELO2 was required for elongation of fatty acids up to 24 carbons, and ELO3 was required for elongation of the 24 carbon fatty acid to 26 carbons. However, neither gene was essential for viability.
  • elegans like most other animals, and in contrast to higher plants, synthesises PUFAs such as arachidonic acid (AA; 20:4 A 5,8 11,14 ) as precursors to a class of molecules known as the eicosanoids, which in turn serve as precursors for compounds such as prostaglandins and leucotrienes (Horrobin, (1990), Reviews in Contemp Pharmacotherpy, 1:1-45).
  • AA arachidonic acid
  • eicosanoids which in turn serve as precursors for compounds such as prostaglandins and leucotrienes
  • Horrobin (1990), Reviews in Contemp Pharmacotherpy, 1:1-45.
  • the presence of AA and other long chain polyunsaturated fatty acids in C. elegans is well documented (Tanaka et al, (1996), Lipids 31, 1173-1178).
  • the complete sequence of the nematode's genome is now publicly available (The C. elegans consortium, 1998, Science
  • An object of the invention is to provide an isolated PUFA elongase.
  • ORFs putative open reading frames
  • a first aspect of the invention provides an isolated polypeptide comprising a functional long chain polyunsaturated fatty acid (PUFA) elongase i.e. the polypeptide has the function of extending the chain length of an 18 carbon PUFA to 20 carbons in length.
  • PUFA long chain polyunsaturated fatty acid
  • the polypeptide may be from a eukaryote.
  • the polypeptide may comprise at least a portion of the amino acid shown in SEQ LD. 15, or variants thereof.
  • variants in relation to a certain sequence means a protein or polypeptide which is derived from the sequence through the insertion or deletion of one or more amino acid residues or the substitution of one or more animo acid residues with amino acid residues having similar properties, e.g. the replacement of a polar amino acid residue with another polar amino acid residue, or the replacement of a non-polar amino acid residue with another non-polar amino acid residue.
  • variants must have an elongase function as defined herein.
  • a second aspect of the invention provides a polypeptide having at least 60 % homology to a polypeptide according to a first aspect of the invention.
  • the polypeptide may have at least 80%), or as much as 90% or more homology to a polypeptide according to a first aspect of the invention.
  • the polypeptide according to either aspect of the invention may include a sequence motif responsible for Endoplasmic Reticulum (ER) - retention. This allows the polypeptide to be specifically located or targeted to the ER of a cell.
  • ER Endoplasmic Reticulum
  • the polypeptide may also be able to elongate palmitoleic acid (PA; 16:1 ⁇ 9 ) to vacceric acid (VA; 18:l ⁇ n ).
  • PA palmitoleic acid
  • VA vacceric acid
  • the polypeptide is also capable of elongation of a ⁇ 9 - monounsaturated 16C fatty acid.
  • the polypeptide is from an animal, more preferably, the animal is an invertebrate such as a worm. Where the animal is a worm, it is preferably C. elegans. Alternatively, the animal is a vertebrate, preferably a mammal such as a human, rat or mouse.
  • a third aspect of the invention provides an isolated DNA sequence, preferably a cDNA sequence, encoding a polypeptide according to a first or second aspect of the invention. This DNA sequence may be used to engineer transgenic organisms.
  • the DNA sequence comprises the sequence shown in SEQ ID NO: 7 or variants of that sequence due, for example, to base substitutions, deletions, and/or additions.
  • a fourth aspect of the invention provides an engineered organism, such as a transgenic animal, engineered to express a polypeptide according to a first or second aspect of the invention.
  • the engineered organism may be engineered to express elevated levels of the polypeptide, thereby providing a supply of polypeptide at a reduced cost as a reduced number of organisms need be used.
  • the engineered organism is a mammal such as a rat, mouse or monkey.
  • a fifth aspect of the invention provides an engineered organism containing a synthetic pathway for the production of a polypeptide according to a first or second aspect of the invention. This has the advantage of allowing greater control over the production of PUFAs by the pathway by an organism.
  • the pathway may include ⁇ -fatty acid desaturase, and/or ⁇ 6 -fatty acid desaturase.
  • the engineered organism according to a fourth or fifth aspect of the invention may be a lower eukaryote, such as yeast.
  • the transgenic organism may be a fish.
  • a sixth aspect of the invention provides a transgenic plant engineered to express a polypeptide according to a first aspect of the invention.
  • a seventh aspect of the invention provides a transgenic plant containing a DNA sequence according to a third aspect of the invention.
  • An eighth aspect of the invention provides a method of producing a PUFA comprising carrying out an elongase reaction catalysed by a polypeptide according to a first or second aspect of the invention.
  • the PUFA may be di-homo-gamma-linoleic acid (20:3 ⁇ 8 11 14 ), arachidonic acid (20:4 ⁇ SA, U4 ), eicosapentanoic acid (20:5 ⁇ 5A11 14 17 ), docosatrienoic acid (22:3 ⁇ 3 16 19 ), docosatetraenoic acid (22:4 ⁇ 7,10 ' 13 ' 16 ), docosapentaenoic acid (22:5 ⁇ 7 10 ' 13 ' 16 ' 19 ) or docosahexaenoic acid (22:6 ⁇ 4 ' 7 - 10 13 16 19 ).
  • the PUFA may be a 24 carbon fatty acid with at least 4 double bonds.
  • a ninth aspect of the invention provides a PUFA produced by a method according to an eighth aspect of the invention.
  • the PUFA may be used in foodstuffs, dietary supplements or pharmaceutical compositions.
  • a tenth aspect of the invention provides a foodstuff comprising a PUFA according to a fifth aspect of the invention.
  • the foodstuff can be fed to an animal.
  • An eleventh aspect of the invention provides a dietary supplement comprising a PUFA according to a fifth aspect of the invention.
  • the dietary supplement can be supplied to an animal to augment its PUFA levels.
  • An twelfth aspect of the invention provides a pharmaceutical composition comprising a polypeptide according to a first or second aspect of the invention or a PUFA according to a ninth aspect of the invention.
  • the pharmaceutical composition comprises a pharmaceutically-acceptable diluent, carrier, excipient or extender.
  • a topical application would preferably be a cream or lotion, whereas if the composition was to be ingested a different form would be more suitable.
  • a thirteenth aspect of the invention provides a method of treatment of an animal, such as a mammal, or a plant, comprising supplying to the animal or plant a DNA sequence according to a third aspect of the invention, a foodstuff according to a tenth aspect of the invention, a dietary supplement according to an eleventh aspect of the invention, a pharmaceutical composition according to a twelfth aspect of the invention or a PUFA according to a ninth aspect of the invention.
  • the mammal is a human.
  • SEQ LDl to 8 show the putative ORFs encoding PUFA elongases A to H respectively;
  • SEQ ID9 to 16 show the deduced amino acid sequences of the putative ORFs of SEQ ID NO: 1 to 8 respectively;
  • Figures 2 to 9 show hydrophobicity plots for each of PUFA elongases A to H respectively.
  • Figure 10 shows an amino acid sequence line-up comparing the C. elegans ORF F56H1 1.4 (Z68749) with related sequences.
  • Figure 11 shows chromatograms of fatty acid methyl esters from transformed yeast.
  • C. elegans databases were searched for any sequences which showed low levels of homology to yeast ELO genes (EL02 and EL03) using the TBLASTN programme.
  • a similar search was carried out using short (20 to 50 amino acid) stretches of ELO genes which were conserved amongst the three ELO polypeptide sequences.
  • C. elegans sequences which were identified by this method were then used themselves as search probes, to identify any related C. elegans genes which the initial search with the yeast sequences failed to identify. This was necessary because the level of homology between the yeast ELO genes .and any worm genes is always low (see BLAST scores later).
  • GenBank is the NIH genetic sequence database, an annotated collection of all publicly available DNA sequences (Nucleic Acid Research (1998) 26, 1-7). There are approximately 2,162,000,000 bases in 3,044,000 sequence records as of December 1998.
  • Wormpep contains predicted proteins from the Caenorhabditis elegans genome sequence project, which is carried out jointly by the Sanger Centre in Cambridge, UK and Genome Sequencing Center in St. Louis, USA.
  • the current Wormpep database, Wormpep 16 contains 16,332 protein sequences (7,120,115 residues). Search strings used included [HXXHH], [HXXXHH], [QXXHH] and [YHH]. Comparison of the data from the two different searches indicated a small ( ⁇ 10) number of putative ORFs as candidate elongases.
  • the histidine box motifs are shown in bold in SEQ ID 9 to 16.
  • fatty acid elongases are expected to be endoplasmic reticulum (ER) membrane proteins, they might be expected to have peptide signals which are responsible for "ER-retention".
  • this signal often takes the form of a C-terminal motif [K-K-X 2 -3-Stop], or similar variants thereof (Jackson et al, (1990), EMBO J., 9, 3153-3162). Further sequence analysis of the C.
  • C. elegans genes involved in the synthesis of PUFA may exist in tandem (for example the ⁇ 5 and ⁇ 6 desaturases required for AA and GLA synthesis, respectively, are ⁇ 1 kB apart on chromosome IN (Michaelson et al, (1998), FEBS Letts 439, 215-218), the positions of the putative C. elegans elongase ORFs were determined using the Sanger Centre's WebAce C. elegans server (http://www.sanger.ac.uk/Projects/C_elegans/webace_front_end.shtml). This indicated that two pairs of putative elongases were in close proximity to each other on the C. elegans chromosome IV.
  • F41H10.7 and F41H10.8 were identified as being approximately 10 Kb apart on chromosome IV, and F56H11.3 and F56H11.4 were identified as being approximately 2 Kb apart on chromosome IV.
  • the positions of the putative ORFs in the C. elegans genome are shown below i.e. chromosome number, and map position in centiMorgans, together with the GenBank database accession numbers.
  • ORF C40H1.4 is predicted coding sequence 4 on cosmid C40H1.
  • Each of the three yeast ELO polypeptides were compared against all of the worm putative elongase tr.anslated ORF sequences, and then ranked in order of similarity (as measured by the BLAST score) (Altschul et al (1990), supra)
  • Yeast ELO 1 (14 to 16 carbon fatty acid elongase)
  • C. elegans ⁇ 5 and ⁇ 6 fatty acid desaturases have evolved from 1 ancestral gene (Michaelson et al, (1998), FEBS Letts 439, 215-218). It is also significant that one pair of C. elegans putative elongase ORFs (F & G) genetically maps close to the ⁇ 5/ ⁇ 6 fatty acid desaturase genes, with both gene pairs being located at the top end of chromosome IV.
  • F56H11.4 and F41H10.8 were cloned by PCR into the pYES2 vector (Invitrogen).
  • a C. elegans mixed stage cDNA library was used as a PCR template.
  • F56H11.4 was amplified using primers:
  • Amplified sequences were then restricted using Kpnl and BamHI (underlined in the forward and reverse primers, respectively), purified using the Qiagen PCR purification kit, and ligated into a Kpnl/BarnHI cut pYes2 vector.
  • Elongases and desaturase constructs were introduced in Saccharomyces cerevisiae W303-1A using a lithium acetate based method (Elble, R. (1992) Biotechniques 13, 18-20) and expression of the transgenes was induced by addition of galactose to 2% (w/v) as described in Napier et al (Napier, J. A., et al (1998) Biochem J 330, 611-614; Michaelson L. V., supra; Michaelson, L. V., (1998) FEBS Letts 439, 215-218).
  • Yeast transformants containing pYES2-derived constructs were grown on synthetic minimal media (SD, the composition of which is defined in Sherman, F (1991) Methods in Enzymology 194, 3-21); synthetic minimal medium minus uracil; pESC-derived constructs were grown on SD minimal medium minus tryptophan.
  • Co-transformed yeast (containing both pYES2 and pESC derivatives) were grown on SD minimal medium minus uracil and tryptophan.
  • cultures Prior to induction, cultures were grown in the presence of 2% raffmose and supplemented with 0.5 mM of the appropriate fatty acid substrate in the presence of 1% tergitol-(NP40) (Sigma). All cultures were then grown for a further 48-h unless indicated.
  • Lipids were extracted from transformed and control yeast by homogenisation in MeOH-CHCl 3 using a modification of the method of Bligh and Dyer (Dickenson & Lester (1999) Biochim Biophys Acta 1426, 347-357).
  • the resulting CHCL phase was evaporated to dryness under nitrogen gas and the samples were transmethylated with 1M HC1 in methanol at 80 °C for 1 hour.
  • Fatty acid methyl esters (FAMES) were extracted in hexane and purified using a small column packed with Florisil.
  • fatty acids extracted from yeast cultures were analysed by gas chromatography (GC) of methyl ester derivatives. Lipids were extracted, transmethylated and the fatty acid methyl esters (FAMEs) analysed as described by Sayanova et al.
  • GC gas chromatography
  • Figure 11 shows chromatograms of fatty acid methyl esters from yeast transformed with the control (empty) plasmid pYES2 (Fig. 11 A) or with ORF F56H11.4 in pYES2 (Fig. 11B). Exogenous substrate in the form of GLA was supplied to the cultures. Two novel peaks are observed in (B); these peaks (annotated as 20:3 and 18: 1 *) were identified (against known standards) as DHGLA and vaccenic acid, respectively. Detection was by flame ionisation.
  • One cDNA ORF tested in this manner displayed a high level of elongase activity on the GLA substrate, converting 44% to DHGLA.
  • the identity of this elongation product was confirmed as DHGLA by comparison with a known standard (the standards used were known standards for either DHGLA, AA, EPA or VA from Sigma Chemicals, Ltd.), using GCMS analysis using a Kratos MS80RFA (Napier, J. A., supra; Michaelson, L. V., supra; Michaelson, L. V., supra).
  • the deduced amino acid sequence of the functional elongase clone identified it as being encoded by the C.
  • elegans gene F56H11.4, and comparison with the yeast ELO genes showed low homology confined to a few short amino acid motifs (see Fig. 10).
  • Some similarity with a mouse gene Cig30 (Tvrdik, P., (1997) J. Biol. Chem. 272, 31738-31746), which has been implicated in the recruitment of brown adipose tissue in liver tissue, was also observed, as well as a potential human homologue encoded by a gene located on chromosome 4q25, BAC 207d4. The most closely related C.
  • elegans ORFs F41H10.8 (U61954) and F56H11.3 (Z68749) are also shown, as is part of a related human gene present on chromosome IV (present on BAC clone B207d4; AC004050).
  • GenBank accession numbers are given for all sequences.
  • the range of fatty acids synthesised by C. elegans can potentially require a number of different elongation reactions (Tanaka, T., (1996) Lipids 31, 1173-1178).
  • the substrate-specificity of the F56H11.4 PUFA elongase was therefore determined using a range of exogenous ly supplied fatty acids. This revealed that GLA is the major substrate, with a number of other fatty acids being elongated at a lower efficiency (see Table 1). Although most of these substrates are polyunsaturated fatty acids, it was unexpectedly observed that palmitoleoic.
  • PA 16:1 ⁇ 9
  • VA vaccenic acid
  • the C. elegans PUFA elongase ORF F56H11.4 maps to the top of chromosome IV (at 4.32 cM) with a related sequence (F56H11.3; 51 % similarity) located l,824bp downstream.
  • Another C. elegans gene (F41H10.8) was also observed, which is present on chromosome IN, and which shows a slightly higher level (53%) of similarity to the PUFA elongase than F56H11.3 (see Fig. 10).
  • the PUFA elongase F56H11.4 was expressed in yeast in conjunction with either the ⁇ 6 - or ⁇ 5 -fatty acid desaturases previously isolated and characterised by the inventor (Napier, J. A., supra; Michaelson, L. V., supra). Expression of the ⁇ 6 -fatty acid desaturase and F56H1 1.4 was carried out in the presence of two different substrates (LA or ALA) while the ⁇ -fatty acid desaturase and the elongase were expressed in the presence of GLA only.
  • DHGLA is an n-6 fatty acid
  • OTA-derived eicostetraenoic acid is an n-3 type
  • Verification was also provided that the 20C PUFAs synthesised in the yeast expression system were generated by the ⁇ 6 -desaturation of 18C substrates which were subsequently elongated, as the ⁇ 6 -desaturase showed no activity on 20:2 or 20:3 substrates (see Table HI).
  • C. elegans ORF sequence can be subcloned into a plant expression vector pJD330, which comprises a viral 35S promoter, and a Nos terminator.
  • the resulting cassette or promoter/coding sequence/terminator can then be subcloned into the plant binary transformation vector pBin 19, and the resulting plasmid introduced into Agrobacterium tumefaciens.
  • This Agrobacterium strain can then be used to transform Arabidopsis by the vacuum-infiltration of inflorescences, and the seeds harvested and plated onto selective media containing kanamycin.
  • Fatty acid methyl ester analysis can be carried out as previously described.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
EP00911091A 1999-03-18 2000-03-20 Elongase für mehrfach ungesättigte fettsäuren aus caenorhabditis elegans Withdrawn EP1161542A1 (de)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
GBGB9906307.5A GB9906307D0 (en) 1999-03-18 1999-03-18 Novel polypeptides
GB9906307 1999-03-18
GB0003869A GB0003869D0 (en) 2000-02-18 2000-02-18 Elongase II
GB0003869 2000-02-18
PCT/GB2000/001035 WO2000055330A1 (en) 1999-03-18 2000-03-20 Polysaturated fatty acid (pufa) elongase from caenorhabditis elegans

Publications (1)

Publication Number Publication Date
EP1161542A1 true EP1161542A1 (de) 2001-12-12

Family

ID=26243677

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00911091A Withdrawn EP1161542A1 (de) 1999-03-18 2000-03-20 Elongase für mehrfach ungesättigte fettsäuren aus caenorhabditis elegans

Country Status (6)

Country Link
EP (1) EP1161542A1 (de)
JP (1) JP2002538826A (de)
AU (1) AU3308500A (de)
CA (1) CA2365096A1 (de)
NO (1) NO20014542L (de)
WO (1) WO2000055330A1 (de)

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6403349B1 (en) * 1998-09-02 2002-06-11 Abbott Laboratories Elongase gene and uses thereof
GB0107510D0 (en) 2001-03-26 2001-05-16 Univ Bristol New elongase gene and a process for the production of -9-polyunsaturated fatty acids
WO2002098889A2 (en) * 2001-06-05 2002-12-12 Exelixis Inc. MAP3Ks AS MODIFIER OF THE p53 PATHWAY AND METHODS OF USE
US7705202B2 (en) 2002-03-16 2010-04-27 The University Of York Transgenic plants expressing enzymes involved in fatty acid biosynthesis
BRPI0413073A (pt) * 2003-08-01 2006-10-03 Basf Plant Science Gmbh processo para a produção de compostos, óleo, lipìdeo ou ácido graxo, ou uma fração dos mesmos, composição de óleo, lipìdeo ou ácido graxo, processo para a produção de óleos, lipìdeos ou composições de ácido graxo, uso de óleo, lipìdeo ou ácidos graxos ou de composições de óleo, lipìdeo ou ácido graxo ou de óleos, lipìdeos ou composições de ácido graxo, seqüência de ácido nucleico isolado, seqüência de aminoácidos, construção de gene, vetor, e, organismo não humano transgênico
US7807849B2 (en) 2004-04-22 2010-10-05 Commonwealth Scientific And Industrial Research Organisation Synthesis of long-chain polyunsaturated fatty acids by recombinant cells
CA2563875C (en) 2004-04-22 2015-06-30 Commonwealth Scientific And Industrial Research Organisation Synthesis of long-chain polyunsaturated fatty acids by recombinant cells
BRPI0511236A (pt) * 2004-06-04 2007-11-27 Fluxome Sciences As método para a produção de ácidos graxos poliinsaturados com quatro ou mais ligações duplas, saccharomyces cerevisiae geneticamente modificado que tem capacidade de produzir ácidos graxos poliinsaturados com quatro ou mais ligações duplas quando desenvolvido em um substrato que não de ácido graxo, composição, uso da composição e uso de um saccharomyces cervisiae geneticamente modificado
WO2008025068A1 (en) 2006-08-29 2008-03-06 Commonwealth Scientific And Industrial Research Organisation Synthesis of fatty acids
US7892792B2 (en) 2008-06-27 2011-02-22 Indian Institute Of Science Cells expressing Pichia cytochrome C
BR122021003835B1 (pt) 2008-11-18 2022-02-08 Grains Research And Development Corporation Polinucleotídeo isolado e/ou exógeno que não ocorre de forma natural, vetor compreendendo o referido polinucleotídeo e método de produção de óleo contendo ácidos graxos insaturados
SG11201408362SA (en) 2012-06-15 2015-01-29 Commw Scient Ind Res Org Production of long chain polyunsaturated fatty acids in plant cells
CN111154724B (zh) 2013-12-18 2024-02-06 联邦科学技术研究组织 包含二十二碳六烯酸的提取的植物脂质
WO2015196250A1 (en) 2014-06-27 2015-12-30 Commonwealth Scientific And Industrial Research Organisation Lipid comprising docosapentaenoic acid
CN104920307B (zh) * 2015-06-19 2018-01-26 安徽省农业科学院农业工程研究所 一种适用于单粒子微束装置的秀丽小杆线虫固定方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6403349B1 (en) * 1998-09-02 2002-06-11 Abbott Laboratories Elongase gene and uses thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0055330A1 *

Also Published As

Publication number Publication date
NO20014542D0 (no) 2001-09-18
NO20014542L (no) 2001-09-18
WO2000055330A9 (en) 2002-08-29
JP2002538826A (ja) 2002-11-19
WO2000055330A1 (en) 2000-09-21
CA2365096A1 (en) 2000-09-21
AU3308500A (en) 2000-10-04

Similar Documents

Publication Publication Date Title
Petrie et al. Development of a Brassica napus (Canola) crop containing fish oil-like levels of DHA in the seed oil
WO2000055330A1 (en) Polysaturated fatty acid (pufa) elongase from caenorhabditis elegans
Beaudoin et al. Heterologous reconstitution in yeast of the polyunsaturated fatty acid biosynthetic pathway
Murata et al. Acyl-lipid desaturases and their importance in the tolerance and acclimatization to cold of cyanobacteria.
Jin et al. Characterization and temporal expression of a ω-6 fatty acid desaturase cDNA from sesame (Sesamum indicum L.) seeds
Voelker et al. Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants
KR102386838B1 (ko) 장쇄 다중불포화 지방산을 포함하는 지질
Zank et al. Cloning and functional characterisation of an enzyme involved in the elongation of Δ6‐polyunsaturated fatty acids from the moss Physcomitrella patens
CA2519169C (en) Transgenic plants expressing enzymes involved in fatty acid biosynthesis
US7041872B2 (en) Acyl CoA:cholesterol acyltransferase related nucleic acid sequences
US6489461B1 (en) Nucleic acid sequences encoding proteins involved in fatty acid beta-oxidation and methods of use
US6426447B1 (en) Plant seed oils
CA2881252C (en) Soybean oil having 5 to 50% sda and less than 10% gla
Froman et al. ACX3, a novel medium-chain acyl-coenzyme A oxidase from Arabidopsis
CA2399349A1 (en) Novel elongase gene and method for producing multiple-unsaturated fatty acids
AU2012286816A1 (en) Production of DHA and other LC-PUFAs in plants
Bryant et al. ACYL-ACYL CARRIER PROTEIN DESATURASE2 and 3 are responsible for making omega-7 fatty acids in the Arabidopsis aleurone
Li et al. A 3-ketoacyl-CoA synthase 11 (KCS11) homolog from Malania oleifera synthesizes nervonic acid in plants rich in 11Z-eicosenoic acid
Claver et al. Functional analysis of β-ketoacyl-CoA synthase from biofuel feedstock Thlaspi arvense reveals differences in the triacylglycerol biosynthetic pathway among Brassicaceae
Guan et al. RNAi targeting putative genes in phosphatidylcholine turnover results in significant change in fatty acid composition in Crambe abyssinica seed oil
CA2343969A1 (en) Novel plant acyltransferases
Lee et al. Lesquerella FAD3-1 gene is responsible for the biosynthesis of trienoic acid and dienoic hydroxy fatty acids in seed oil
Kim et al. Functional characterization of polyunsaturated fatty acid delta 6-desaturase and elongase genes from the black seabream (Acanthopagrus schlegelii)
Park et al. Enhanced production of hydroxy fatty acids in Arabidopsis seed through modification of multiple gene expression
NL9002130A (nl) Dna-sequentie die ten minste een deel van een gen coderend voor stearoyl-acp-desaturase omvat, alsmede toepassing van de dna-sequentie in een werkwijze voor het wijzigen van de vetzuurbiosynthese van een plant.

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20011005

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

AX Request for extension of the european patent

Free format text: AL;LT;LV;MK;RO;SI

17Q First examination report despatched

Effective date: 20050531

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20051213