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WO1994011519A1 - Production de polyhydroxyalcanoate dans des plantes - Google Patents

Production de polyhydroxyalcanoate dans des plantes Download PDF

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Publication number
WO1994011519A1
WO1994011519A1 PCT/GB1993/002286 GB9302286W WO9411519A1 WO 1994011519 A1 WO1994011519 A1 WO 1994011519A1 GB 9302286 W GB9302286 W GB 9302286W WO 9411519 A1 WO9411519 A1 WO 9411519A1
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Prior art keywords
plant
genes
pha
polyhydroxyalkanoate
production
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PCT/GB1993/002286
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English (en)
Inventor
Philip Anthony Fentem
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Zeneca Limited
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Publication date
Application filed by Zeneca Limited filed Critical Zeneca Limited
Priority to AU54271/94A priority Critical patent/AU5427194A/en
Publication of WO1994011519A1 publication Critical patent/WO1994011519A1/fr

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    • 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.)
    • C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
    • 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
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • 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)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C12P7/625—Polyesters of hydroxy carboxylic acids

Definitions

  • This invention relates to the production of polyhydroxyalkanoate in plants.
  • Poly-3-hydroxybutyrate is a linear polyester of D( - )-3-hydroxybutyrate . It was first discovered in Bacillus megaterium in 1925. Polyhydroxybutyrate accumulates in intracellular granules of a wide variety of bacteria. The granules appear to be membrane bound and can be stained with Sudan Black dye. The polymer is produced under conditions of nutrient limitation and acts as a reserve of carbon and energy. The molecular weight of the polyhydroxybutyrate varies from around 50,000 to greater than 1,000,000, depending on the micro-organisms involved , the conditions of growth, and the method employed for extraction of the polyhydroxybutyrate.
  • Polyhydroxybutyrate is an ideal carbon reserve as it exists in the cell in a highly reduced state, (it is virtually insoluble), and exerts negligible osmotic pressure.
  • Polyhydroxybutyrate and related poly-hydroxy- alkanoates such as poly-3-hydroxyvalerate and poly-3-hydroxyoctanoa e , are biodegradable thermo-plastics of considerable commercial importance.
  • polyhydroxyalkanoates and "PHA” as used hereinafter includes polymers of 3-h ydr oxybut yra te , polymers of related hydroxyalkanoates such as 3-hydroxyvalerate, 3-hydroxyhexanoate , 3-hydroxyoctanoate , 3-hydroxydecanoate, and also copolymers containing more than one of these hydroxyalkanoates.
  • Polyhydroxyalkanoate is biodegradable and is broken down rapidly by soil micro-organisms. It is thermoplastic (it melts at 180°C) and can readily be moulded into diverse forms using technology well-established for the other thermo-plastics materials such as high-density polyethylene which melts at around the same temperature (190°C). The material is ideal for the production of biodegradable packaging which will degrade in landfall sites and sewage farms.
  • the polymer is biocompatible, as well as biodegradable, and is well tolerated by the mammalian, including human, body,its degradation product, 3-hydroxybutyrate, is a normal mammalian metabolite.
  • polyhydroxyalkanoate degrades only slowly in the body and its medical uses are limited to those applications where long term degradation is required.
  • Polyhydroxyalkanoate produced by the micro-organism Alcaligenes eutrophus , is manufactured, as a copolymer of polyhydroxybutyrate and polyhydroxyvalerate, by Imperial Chemical Industries PLC and sold under the Trade Mark BIOPOL. It is normally supplied in the form of pellets for thermoprocessing.
  • polyhydroxyalkanoate is more expensive to manufacture by existing methods than, say, polyethylene. It is, therefore, desirable that new, more economic production of polyhydroxyalkanoate be provided.
  • An object of the present invention is to provide materials and a method for the efficient production of polyhydroxyalkanoate.
  • a plant adapted for the intracellular production of polyhydroxyalkanoate comprising a recombinant genome of an oil-producing plant, which genome contains genes encoding enzymes necessary for catalysing the production of PHA together with gene regulatory sequences directing expression of the said genes to target plant cell components, expression of the said genes being controlled by a promoter sequence which is inducible by the application of a chemical inducer.
  • PHA polyhydroxyalkanoate
  • genes encoding the enzyme or enzymes necessary for the catalysis of polyhydroxyalkanoate production may be isolated from a micro-organism such as Alcaligenes eutrophus, Rhodococcus ruber, Pseudomonas oleovorans or Pseudomonas aeruginosa.
  • the preferred oil-producing plant is selected from the group consisting of oilseed rape, canola, soya and sunflower.
  • the said enzymes for catalysing PHA production are ⁇ -ketothiolase, acetoacetyl-CoA-reductase , polyhydroxy-alkanoate- synthetase and 3-hydroxyalkanoyl
  • the said gene regulatory sequences preferably direct expression of the polyhydroxyalkanoate genes specifically to the germinating oilseed, for example the germinating embryo.
  • the said chemically inducible promoter sequence may be the promoter of the gene specifying the 27kd sub-unit of the glutathione-S-transferase enzyme in Zea mays .
  • the said gene regulatory sequences direct expression of the polyhydroxyalkanoate genes to the cytosol or to the glyoxysome.
  • regulatory sequences include promoter sequences directing expression of the biosynthetic pathway specifically to the germinating seed, and transit peptide sequences targeting the enzymes to appropriate subcellular compartments.
  • the plant be of a species which produces substantial quantities of oil, rather than starch.
  • Such plant species are well known and are simply referred to as "oil-seed” crops and include, oilseed rape, canola, maize, soya and sunflower.
  • Alcaligenes eutrophus from the substance, acetyl-CoA involves three enzyme-catalysed steps, illustrated in Figure 1 herewith.
  • the three enzymes involved are B-ketothiolase, NADP linked acetoacetyl CoA reductase and polyhydroxybutyrate synthase, the genes for which have gene cloned from Alcaligenes eutrophus fSchubert et al,. J. Bacteriol. Vol 170 (1988)].
  • B-ketothiolase NADP linked acetoacetyl CoA reductase
  • polyhydroxybutyrate synthase the genes for which have gene cloned from Alcaligenes eutrophus fSchubert et al,. J. Bacteriol. Vol 170 (1988)].
  • the three genes are known to facilitate production of polyhydroxyalkanoate up to 30% of the cell weight.
  • An important preferred feature of this invention is the use of an oilseed plant for expression of the polyhydroxyalkanoate.
  • a feature of this invention is that expression of polyhydroxyalkanoate (PHA) polymers is obtained in the germinating seed of oilseed crops. Germinating oilseeds produce large amounts of acetyl CoA, the substrate required for PHA synthesis, in the cotyledons as a result of fatty acid oxidation, which occurs during the mobilisation of lipid reserves. PHA would be produced by harvesting the seed of oilseed crop plants transformed with PHA biosynthetic genes from bacterial sources. Then seed would then be germinated in a process akin to malting. During the malting process the seed would produce PHA polymers which can then be extracted.
  • PHA polyhydroxyalkanoate
  • the invention provides a "dual-purpose" crop.
  • the product seed is simply ordinary oilseed.
  • the PHA producing genes are switched on and the crop produces PHA.
  • the crop may be harvested as an ordinary oilseed crop and used as such or, should circumstances dictate, the seed may be malted and PHA produced in the malting process .
  • the acetyl CoA produced by the oilseed during germination is normally destined for respiration, to produce energy and CO,, or destined for synthesis of sucrose, a process known as gluconeogenesis , via the glyoxylate pathway.
  • the sucrose is then transported to the growing points of the germinating embryo.
  • a feature of this invention is that the acetyl CoA is diverted from this glyoxylate pathway into, the production of PHA polymers.
  • Rhodococcus ruber PHA synthase can utilise (+) isomers of hydroxyalkanoyl CoAs as substrates .
  • L( + )-3-hydroxybutyryl CoA is produced as an intermediate of fatty acid oxidation by germinating oilseeds.
  • PHA polymers could be produced in germinating oilseeds by gaining expression of the PHA synthase of Rhodococcus ruber, or expression of PHA synthases with similar substrate specificities from other organisms, iv) L(+ )-3-hydroxyalkanoyl CoAs of chain length C i to C j g are produced as intermediates of fatty acid oxidation in germinating oilseeds.
  • PHA synthase genes of these organisms have been isolated. Expression of these Pseudomonas oleovorans genes in germinating oilseeds therefore could enable these tissues to synthesise PHA polymers directly from the intermediates of fatty acid oxidation. It is possible that polymer synthesis in Pseudomonas oleovorans involves the initial conversion of L(+) hydroxyalkanoyl CoAs to their D(-) isomers via the action of two stereospecific dehydratase enzymes. In this case, expression of such PHA polymers in germinating oilseeds would require expression of these two dehydratases genes in addition to the PHB synthase gene. For expression in higher plants the bacterial genes require suitable promoter and terminator sequence.
  • the cauliflower mosaic virus CaMV35S promoter and nopaline synthase (nos) terminator may be used for constitutive expression. It is however preferred to target synthesis of PHA only to the germinating oilseed eg, the germinating embryo of oilseed rape.
  • a feature of this invention is to target production of PHA specifically to the germinating seed. One way of achieving this is to make gene constructs for all the necessary PHA synthetic genes containing a promoter sequence which directs expression specifically to the seed, and specifically to the period of germination.
  • Such a promoter may be obtained by: i) Differential hybridisation screening of cDNA libraries obtained from seed at an early stage of germination, with libraries obtained from developing seed, leaf, root and germinating seedlings at a later stage of germination. This approach should yield a gene, and thus a promoter, expressed specifically during the early stage of germination, ii) Isolation of the promoter sequence of the gene encoding the enzyme lipase. Lipase enzymes are expressed specifically at the early stage of germination.
  • Isolation of the gene requires purification of the protein, obtaining amino acid sequences from peptides derived from the purified protein, using these sequences to reverse clone a cDNA and genomic DNA for lipase by techniques well known to those acquainted with the art. iii) Isolation of the promoter sequence of a gene for a protease. Serine and cysteine proteases are known to be expressed at an early stage of germination, . conserveed sequence elements have been described for such proteases isolated from a range of organisms.
  • These common sequence elements can be used to amplify a portion of a gene for a protease expressed specifically during seed germination using PCR, with a cDNA library from germinating seed as template, and the common sequence elements as primers.
  • the PCR product could then be used to isolate a full length cDNA and genomic DNA for the protease, from which the promoter could be isolated.
  • a second way of targeting PHB production specifically to the germinating seed is to make gene constructs for all the necessary PHA synthetic genes containing a promoter sequence from a gene which is chemically inducible.
  • a promoter is that of the glutathione-S-methyl transferase II (GSTII) gene isolated from Zea mays . Expression of this gene is induced by chemicals known as safeners. If PHA biosynthetic genes are placed under the control of this promoter PHA polymers will not normally be produced in oilseed plants tissues. Seed will be produced normally and harvested. A chemical safener compound is then applied to the seed, either as a seed dressing or sprayed onto the seed. The seeds are then germinated (“malted”) by incubating in the presence of water, and the germinating seed would then express the PHA biosynthetic genes and produce PHA.
  • GSTII glutathione-S-methyl transferase II
  • the key pathway producing acetyl CoA in germinating oilseed is fatty acid oxidation. While a small proportion (3%) of fatty oxidation occurs in mitochondria, the major proportion (97%) occurs in the glyoxysome, where the acetyl CoA product is normally used to produce succinate via the "glyoxylate pathway". The succinate produced then enters into the gluconeogenic pathway involving reactions in both mitochondria and cytosol, culminating in the production of sucrose. While significant storage space for PHA polymers exists in the glyoxysomes , potentially greater storage space exists in the cytosol.
  • the acetyl CoA generated in the glyoxysome could give rise to acetyl CoA in the cytosol via glyoxysomal citrate synthase, outward transport of citrate to the cytosol, and cleavage of citrate via cytosolic citrate lyase, yielding acetyl CoA. It may be possible to obtain PHB synthesis in germinating oilseed by gaining expression of the three PHB biosynthetic pathway enzymes form Alcaligenes eutrophus in the cell cytosol.
  • acetyl CoA in the cytosol does ' not occur at a sufficiently high rate however, it will be necessary to site the Alcaligenes eutrophus ketothiolase enzyme in the glyoxysome to compete directly with glyoxylate cycle enzymes for the acetyl CoA produced by fatty acid oxidation. If the acetoacetyl CoA product produced by this enzyme can not traverse the glyoxysomal envelope membrane, it will be necessary to target the Alcaligenes NADP acetoacetyl CoA reductase to the glyoxysome as well.
  • the 3-hydroxybutyryl CoA intermediate can not traverse the glyoxysome envelope membrane, it will be necessary to target PHB synthase to the glyoxysome, and achieve PHB synthesis, inside this organelle. If the acetoacetyl CoA or 3-hydroxybutyryl CoA intermediates can however traverse the glyoxysome envelope membrane, it will be possible to express ketothiolase in the glyoxysome and PHB polymerase (and possibly acetoacetyl CoA reductase) in the cytosol, in order to achieve acetyl CoA utilisation in the glyoxysome and PHB synthesis in the cytosol .
  • acetoacetyl CoA produced as an intermediate of fatty acid oxidation, for synthesis of PHB directly, without requiring expression of the Alcaligenes ketothiolase gene.
  • acetoacetyl CoA or 3-hydroxybutyryl CoA can be transported from the glyoxysome to the cytosol, it may be necessary to express the Alcaligenes NADP acetoacetyl CoA reductase and PHB synthase in the glyoxysome or cytosol to achieve maximal PHB synthesis.
  • L( +)-3-hydroxybutyryl CoA is also generated in the glyoxysome as an intermediate of fatty acid oxidation.
  • a PHB synthase such as that from Rhodococcus ruber
  • L(+)-3-hydroxyalkanoyl CoAs of chain length C ⁇ -C j - are produced in the glyoxysome as intermediates of fatty acid oxidation.
  • Pseudomonas oleovorans capable of converting
  • CoAs CoAs
  • Pseudomonas oleovorans capable of synthesising polymers
  • D( -) -3-hydroxyalkanoyl CoAs of chain length C ⁇ -C ⁇ could also be expressed in the glyoxysome or cytosol dependent on the abilities of L( + ) or D(-) hydroxyalkanoyl CoA intermediates to be exported from the glyoxysome.
  • the best documented peroxisomal targeting sequence is a tripeptide serine-lysine-leucine (SKL) located at the carboxy terminus, or within the carboxy terminus domain of the protein.
  • SSL serine-lysine-leucine
  • the final amino acid of the tripeptide must be leucine .
  • Rat peroxisomal thiolases contain an amino terminal 26 or 36 amino acid sequence which is cleaved from the protein during import into the peroxisome. Deletion analysis has shown the first 11 amino acids of this targeting sequence only to be necessary for import into the peroxisome. The approach therefore is to construct PHA biosynthetic genes containing the amino terminal targeting sequence (11, 26 or 36 amino acids) to gain glyoxysomal expression of the necessary PHA biosynthetic enzymes in germinating oilseeds.
  • Plants are individually transformed with PHA pathway genes. Plants containing individual genes are grown up in the glass house and cross pollinated to obtain hybrid plants containing two pathway genes. This procedure is then repeated to produce hybrid plants containing three or more genes.
  • Plants are sequentially transformed with plasmids containing the individual pathway genes.
  • iii Two or three pathway genes are cotransformed into the same plant by simultaneous infection with Agrobacteria containing the individual genes.
  • Plants are transformed with plasmids containing two or three pathways genes.
  • a combination of these techniques may be used to obtain expression of two ore more genes in a single plants.
  • methods (ii) and (iii) it is advantageous to insert individual genes into vectors containing different selectable marker genes to facilitate selection of plants containing two or more PHA pathway genes.
  • selectable markers are genes conferring resistances to kanamycin, hygromycin, sulphonamides and Basta.

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Abstract

Dans cette invention on place des gènes déterminant la biosynthèse de polyhydroxyalcanoates sous le contrôle d'un promoteur inductible et on les insère dans le génome d'une plante à graines oléagineuses. Lorsque le promoteur est induit par l'application de son inducteur chimique, la plante produit des polyhydroxyalcanoates alors qu'en l'absence de l'inducteur la plante produit des graines oléagineuses ordinaires. Les génomes insérés comprennent de préférence des séquences de ciblage qui dirigent l'expression des gènes vers le cytosol ou le glyoxysome et qui peuvent être exprimées pendant la germination des graines. Si la graine de la plante modifiée est cultivée en l'absence de l'inducteur et récoltée, elle peut être utilisée comme une graine oléagineuse ordinaire ou, selon le choix du producteur, elle peut être soumise à un traitement de maltage en présence de l'inducteur du promoteur pour produire des polyhydroxyalcanoates.
PCT/GB1993/002286 1992-11-06 1993-11-05 Production de polyhydroxyalcanoate dans des plantes WO1994011519A1 (fr)

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AU54271/94A AU5427194A (en) 1992-11-06 1993-11-05 Production of polyhydroxyalkanoate in plants

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GB9223332.9 1992-11-06
GB929223332A GB9223332D0 (en) 1992-11-06 1992-11-06 Production of polyhydroxyalkanoate in plants

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Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995014099A3 (fr) * 1993-11-16 1995-09-08 Univ California Procede de production de proteines dans les plantes
WO1997007230A1 (fr) * 1995-08-21 1997-02-27 The Procter & Gamble Company Extraction par solvant de polyhydroxyalcanoates d'une biomasse
US5610041A (en) * 1991-07-19 1997-03-11 Board Of Trustees Operating Michigan State University Processes for producing polyhydroxybutyrate and related polyhydroxyalkanoates in the plastids of higher plants
US5650555A (en) * 1991-07-19 1997-07-22 Board Of Trustees Operating Michigan State University Transgenic plants producing polyhydroxyalkanoates
WO1997032986A3 (fr) * 1996-03-05 1997-11-20 Weissheimer Friedr Malzfab Procede de production de produits de degradation et/ou de conversion de substances de stockage presentes dans des matieres vegetales transgeniques utilisant un processus de maltage
WO1998006854A1 (fr) * 1996-08-13 1998-02-19 Monsanto Company Sequence d'adn servant a produire des polyhydroxyalcanoates
WO1998000557A3 (fr) * 1996-06-28 1998-07-23 Monsanto Co PROCEDES POUR OPTIMISER DES RESERVES DE SUBSTRATS ET BIOSYNTHESE DE POLY-β-HYDROXYBUTYRATE-CO-POLY-β-HYDROXYVALERATE DANS DES BACTERIES ET DES PLANTES
EP0870053A1 (fr) * 1995-12-19 1998-10-14 Regents Of The University Of Minnesota Procede metabolique de fabrication de synthases monomeres de polyhydroxyalcanoates
WO1999035278A1 (fr) * 1998-01-05 1999-07-15 Monsanto Company Biosynthese de polyhydroxyalcanoates a longueur de chaine moyenne
US5942660A (en) * 1996-03-13 1999-08-24 Monsanto Company Methods of optimizing substrate pools and biosynthesis of poly-β-hydroxybutyrate-co-poly-β-hydroxyvalerate in bacteria and plants
US6083729A (en) * 1995-10-26 2000-07-04 Metabolix, Inc. Methods for isolating polyhydroxyalkanoates from plants
US6175061B1 (en) 1991-04-24 2001-01-16 Monsanto Company Production of polyhydroxyalkanoate in plants
WO2001041559A1 (fr) * 1999-12-10 2001-06-14 Unicrop Ltd Processus de transformation de reserves de stockage de graines de dicotyledone en compositions comprenant un ou plusieurs produits geniques
US6316262B1 (en) * 1997-09-19 2001-11-13 Metabolix, Inc. Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
US6528706B1 (en) 1987-06-29 2003-03-04 Massachusetts Institute Of Technology Polyhydroxybutyrate polymerase
US6946588B2 (en) 1996-03-13 2005-09-20 Monsanto Technology Llc Nucleic acid encoding a modified threonine deaminase and methods of use
US7176349B1 (en) 1999-09-29 2007-02-13 Pioneer Hi-Bred International, Inc. Production of polyhydroxyalkanoate in plants
WO2008026632A1 (fr) 2006-09-01 2008-03-06 Kaneka Corporation Composition élastomère thermoplastique
US7341856B2 (en) 1999-09-29 2008-03-11 Pioneer Hi-Bred International, Inc. Production of polyhydroxyalkanoate in plants
US8889399B2 (en) 2007-03-16 2014-11-18 Genomatica, Inc. Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
US9175297B2 (en) 2008-09-10 2015-11-03 Genomatica, Inc. Microorganisms for the production of 1,4-Butanediol
US9222113B2 (en) 2009-11-25 2015-12-29 Genomatica, Inc. Microorganisms and methods for the coproduction 1,4-butanediol and gamma-butyrolactone
US9410167B2 (en) 2005-05-24 2016-08-09 Lg Chem, Ltd. Cells or plants producing polylactate or its copolymers and uses thereof
US9434964B2 (en) 2009-06-04 2016-09-06 Genomatica, Inc. Microorganisms for the production of 1,4-butanediol and related methods
US9677045B2 (en) 2012-06-04 2017-06-13 Genomatica, Inc. Microorganisms and methods for production of 4-hydroxybutyrate, 1,4-butanediol and related compounds

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332104A2 (fr) * 1988-03-08 1989-09-13 Ciba-Geigy Ag Sèquences d'ADN et gènes chimiquement regulables, et leur emploi
WO1990008826A1 (fr) * 1989-01-26 1990-08-09 Imperial Chemical Industries Plc Interrupteur de gene
EP0388186A1 (fr) * 1989-03-17 1990-09-19 E.I. Du Pont De Nemours And Company Régulation externe de l'expression de gènes
WO1991000917A1 (fr) * 1989-07-10 1991-01-24 Massachusetts Institute Of Technology Procede de production de nouveaux biopolymeres de polyester

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332104A2 (fr) * 1988-03-08 1989-09-13 Ciba-Geigy Ag Sèquences d'ADN et gènes chimiquement regulables, et leur emploi
WO1990008826A1 (fr) * 1989-01-26 1990-08-09 Imperial Chemical Industries Plc Interrupteur de gene
EP0388186A1 (fr) * 1989-03-17 1990-09-19 E.I. Du Pont De Nemours And Company Régulation externe de l'expression de gènes
WO1991000917A1 (fr) * 1989-07-10 1991-01-24 Massachusetts Institute Of Technology Procede de production de nouveaux biopolymeres de polyester

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
POIRIER, Y., ET AL.: "Perspectives on the production of polyhydroxyalkanoates in plants", FEMS MICROBIOLOGY REVIEWS.PAPERS PRESENTED AT THE INTERNATIONAL SYMPOSIUM ON BACTERIAL POLYHYDROXYALKANOATES, FRG, 1-5 JUNE 1992, vol. 103, no. 2-4, 1992, pages 237 - 246 *
POIRIER, Y., ET AL.: "Polyhydroxybutyrate, a biodegradable thermoplastic, produced in transgenic plants", SCIENCE, vol. 256, 24 April 1992 (1992-04-24), LANCASTER, PA US, pages 520 - 523, XP002026974, DOI: doi:10.1126/science.256.5056.520 *
POOL, R.: "In search of the plastic potato", SCIENCE, vol. 245, 15 September 1989 (1989-09-15), LANCASTER, PA US, pages 1187 - 1189, XP002216542, DOI: doi:10.1126/science.245.4923.1187 *

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7109007B2 (en) 1987-06-29 2006-09-19 Massachusetts Institute Of Technology Polyhydroxybutyrate polymerase
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US6175061B1 (en) 1991-04-24 2001-01-16 Monsanto Company Production of polyhydroxyalkanoate in plants
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WO1995014099A3 (fr) * 1993-11-16 1995-09-08 Univ California Procede de production de proteines dans les plantes
US5942597A (en) * 1995-08-21 1999-08-24 The Procter & Gamble Company Solvent extraction of polyhydroxyalkanoates from biomass
WO1997007230A1 (fr) * 1995-08-21 1997-02-27 The Procter & Gamble Company Extraction par solvant de polyhydroxyalcanoates d'une biomasse
US6083729A (en) * 1995-10-26 2000-07-04 Metabolix, Inc. Methods for isolating polyhydroxyalkanoates from plants
US6709848B1 (en) 1995-10-26 2004-03-23 Metabolix, Inc. Methods for isolating polyhydroxyalkanoates from plants
US6600029B1 (en) 1995-12-19 2003-07-29 Regents Of The University Of Minnesota Metabolic engineering of polyhydroxyalkanoate monomer synthases
EP0870053A1 (fr) * 1995-12-19 1998-10-14 Regents Of The University Of Minnesota Procede metabolique de fabrication de synthases monomeres de polyhydroxyalcanoates
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US5958745A (en) * 1996-03-13 1999-09-28 Monsanto Company Methods of optimizing substrate pools and biosynthesis of poly-β-hydroxybutyrate-co-poly-β-hydroxyvalerate in bacteria and plants
US6946588B2 (en) 1996-03-13 2005-09-20 Monsanto Technology Llc Nucleic acid encoding a modified threonine deaminase and methods of use
US5942660A (en) * 1996-03-13 1999-08-24 Monsanto Company Methods of optimizing substrate pools and biosynthesis of poly-β-hydroxybutyrate-co-poly-β-hydroxyvalerate in bacteria and plants
US7192753B2 (en) 1996-03-13 2007-03-20 Monsanto Technology Llc Modified threonine deaminase
WO1998000557A3 (fr) * 1996-06-28 1998-07-23 Monsanto Co PROCEDES POUR OPTIMISER DES RESERVES DE SUBSTRATS ET BIOSYNTHESE DE POLY-β-HYDROXYBUTYRATE-CO-POLY-β-HYDROXYVALERATE DANS DES BACTERIES ET DES PLANTES
USRE37543E1 (en) * 1996-08-13 2002-02-05 Monsanto Company DNA sequence useful for the production of polyhydroxyalkanoates
US5750848A (en) * 1996-08-13 1998-05-12 Monsanto Company DNA sequence useful for the production of polyhydroxyalkanoates
WO1998006854A1 (fr) * 1996-08-13 1998-02-19 Monsanto Company Sequence d'adn servant a produire des polyhydroxyalcanoates
US6689589B2 (en) 1997-09-19 2004-02-10 Metabolix, Inc. Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
US7229804B2 (en) 1997-09-19 2007-06-12 Metabolix, Inc. Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
US6316262B1 (en) * 1997-09-19 2001-11-13 Metabolix, Inc. Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
US7081357B2 (en) 1997-09-19 2006-07-25 Metabolix, Inc. Biological systems for manufacture of polyhydroxyalkanoate polymers containing 4-hydroxyacids
WO1999035278A1 (fr) * 1998-01-05 1999-07-15 Monsanto Company Biosynthese de polyhydroxyalcanoates a longueur de chaine moyenne
US7176349B1 (en) 1999-09-29 2007-02-13 Pioneer Hi-Bred International, Inc. Production of polyhydroxyalkanoate in plants
US7341856B2 (en) 1999-09-29 2008-03-11 Pioneer Hi-Bred International, Inc. Production of polyhydroxyalkanoate in plants
AU782169B2 (en) * 1999-12-10 2005-07-07 Unicrop Ltd A process for converting storage reserves of dicot seeds into compositions comprising one or more gene products
EP1857557A2 (fr) * 1999-12-10 2007-11-21 Unicrop Ltd. Processus de transformation de réserves de stockage de granes de dicotylédone en compositions comprenant un ou plusieures produits géniques
WO2001041559A1 (fr) * 1999-12-10 2001-06-14 Unicrop Ltd Processus de transformation de reserves de stockage de graines de dicotyledone en compositions comprenant un ou plusieurs produits geniques
EP1857557A3 (fr) * 1999-12-10 2009-02-25 Unicrop Oy Processus de transformation de réserves de stockage de granes de dicotylédone en compositions comprenant un ou plusieures produits géniques
US9410167B2 (en) 2005-05-24 2016-08-09 Lg Chem, Ltd. Cells or plants producing polylactate or its copolymers and uses thereof
WO2008026632A1 (fr) 2006-09-01 2008-03-06 Kaneka Corporation Composition élastomère thermoplastique
US8889399B2 (en) 2007-03-16 2014-11-18 Genomatica, Inc. Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
US9487803B2 (en) 2007-03-16 2016-11-08 Genomatica, Inc. Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
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US8969054B2 (en) 2007-03-16 2015-03-03 Genomatica, Inc. Compositions and methods for the biosynthesis of 1,4-butanediol and its precursors
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US11401534B2 (en) 2009-06-04 2022-08-02 Genomatica, Inc. Microorganisms for the production of 1,4- butanediol and related methods
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US10662451B2 (en) 2009-11-25 2020-05-26 Genomatica, Inc. Microorganisms and methods for the coproduction 1,4-butanediol and gamma-butyrolactone
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US9222113B2 (en) 2009-11-25 2015-12-29 Genomatica, Inc. Microorganisms and methods for the coproduction 1,4-butanediol and gamma-butyrolactone
US11085015B2 (en) 2012-06-04 2021-08-10 Genomatica, Inc. Microorganisms and methods for production of 4-hydroxybutyrate, 1,4-butanediol and related compounds
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