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WO2008133398A1 - Procédé d'élaboration d'acide hydroxy-linoléique au moyen de lipoxygénase de maïs et acide hydroxy-linoléique ainsi obtenu - Google Patents

Procédé d'élaboration d'acide hydroxy-linoléique au moyen de lipoxygénase de maïs et acide hydroxy-linoléique ainsi obtenu Download PDF

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Publication number
WO2008133398A1
WO2008133398A1 PCT/KR2008/001049 KR2008001049W WO2008133398A1 WO 2008133398 A1 WO2008133398 A1 WO 2008133398A1 KR 2008001049 W KR2008001049 W KR 2008001049W WO 2008133398 A1 WO2008133398 A1 WO 2008133398A1
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Prior art keywords
acid
lipoxygenase
hydroxyoctadecadienoic
hydroperoxidation
molecules
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PCT/KR2008/001049
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English (en)
Inventor
Ok Soo Han
Sung Kuk Jang
Kyoung Won Cho
Keum Hwa Kim
Eun Ji Um
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Industry Foundation Of Chonnam National University
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Priority claimed from KR1020080012663A external-priority patent/KR100938714B1/ko
Application filed by Industry Foundation Of Chonnam National University filed Critical Industry Foundation Of Chonnam National University
Publication of WO2008133398A1 publication Critical patent/WO2008133398A1/fr

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    • 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
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone

Definitions

  • the present invention relates to a method for preparing hydroxylinoleic acid using maize (Zea mays) lipoxygenase and hydroxylinoleic acid prepared by the same.
  • the present invention relates to a method for preparing hydroxylinoleic acid with controlled distribution of various hydroperoxylinoleic acids from linoleic acid, using maize (Zea mays) lipoxygenase, and hydroxylinoleic acid prepared by the same.
  • Lipoxygenase is a non-heme iron dioxygenase that catalyzes hydroperoxidation of unsaturated fatty acid substrates having a cis, cw-l,4-pentadiene structure. Even though most of prokaryotes including yeast do not harbor a lipoxygenase gene, lipoxygenase is expressed at a high level in animal and plant cells and plays an important role in signal transduction pathways which are mediated by unsaturated fatty acids (Watanabe et al., 1997).
  • Linoleic acid or linolenic acid which is used as a substrate for lipoxygenase is the major unsaturated fatty acid constituting plant membranes, and the membrane fluidity is determined by the content of these unsaturated fatty acids.
  • the membrane fluidity plays a crucial role in adaptation of plants in response to various changes and fluctuations in abiotic environmental variables such as temperature, moisture, salts, and the like and biotic environmental variables such as invasion of pathogenic bacteria, harmful insects, fungi, and the like (Horvath et al., 1983).
  • an intracellular concentration of lipoxygenase substrates and lipoxygenase activity are positioned at the rate-determining step of a conversion process of these unsaturated fatty acids into other physiologically active substances by the lipoxygenase pathway, thereby resulting in formation of a plant stress hormone, e.g. jasmonic acid, as well as a plant wound hormone, e.g. traumatin, and have significant effects on another formation pathway of abscisic acid via violaxanthin.
  • a plant stress hormone e.g. jasmonic acid
  • a plant wound hormone e.g. traumatin
  • 13(S)-hydroperoxy fatty acid and 9(S)-hydroperoxy fatty acid which are produced by the plant lipoxygenase serve as a substrate of hydroperoxide lyase and allene oxide synthase to thereby form the lipoxygenase pathway which will produce various kinds of oxylipins.
  • Oxylipins including jasmonic acid, traumatic acid and ⁇ -ketol are biosynthesized from 13(S)- hydroperoxylinolenic acid which is obtained from linolenic acid, whereas oxylipins including various kinds of alkenals are biosynthesized from linoleic acid.
  • the substrate specificity and positional specificity (or regiospecificity) of lipoxygenase which correspond to a starting point of the lipoxygenase pathway play a key part in what kind of compound will be produced among various kinds of oxylipins having unique physiological activity.
  • Most of lipoxygenases catalyze regioselective and stereoselective reactions, so they may be broadly classified into 9-lipoxygenase that catalyzes hydroperoxidation at the
  • each of these lipoxygenases produces only the corresponding specific stereoisomers. Exceptionally, it was suggested that lipoxygenases found in some plants including maize lipoxygenase exhibit a broad substrate specificity without being regioselective or stereoselective, and therefore produces a variety of reaction products.
  • Korean Patent Application Publication No. 2001-0085101 Al discloses that the maize lipoxygenase concurrently catalyzes 9-hydroperoxidation and 13- hydroperoxidation to produce, as reaction products, 9-hydroperoxylinoleic acid and 13- hydroperoxylinoleic acid which can be used as a starting material of diverse oxylipin biosynthetic pathways of plants.
  • the present invention has been made in view of the above problems, and it is an object of the present invention to provide a method for preparing hydroxylinoleic acid with controlled distribution of various hydroperoxylinoleic acids from linoleic acid, using maize ⁇ Zea mays) lipoxygenase.
  • a method for preparing hydroxylinoleic acid comprising: subjecting linoleic acid to dual regiospecific hydroperoxidation using a mixed solution containing linoleic acid and maize ⁇ Zea mays) lipoxygenase; reducing the resulting reaction product using triphenylphosphine (TPP) or sodium borohydride (NaBH 4 ) as a reducing agent; and separating the resulting reduction product.
  • TPP triphenylphosphine
  • NaBH 4 sodium borohydride
  • the lipoxygenase may be a protein having an amino acid sequence as set forth in SEQ ID NO: 2 or a protein having a sequence homology of more than 70% to the protein having an amino acid sequence of SEQ ID NO: 2.
  • the reaction product may be 13-(9Z,l lE)-hydroperoxyoctadecadienoic acid, 13- (9E, 11 E)-hydroperoxyoctadecadienoic acid, 9-( 1 OE, 12Z)-hydroperoxyoctadecadienoic acid or 9-(10E,12E)-hydroperoxyoctadecadienoic acid.
  • a concentration of linoleic acid may be in the range of 0.03 mM to 0.5 mM.
  • a concentration of lipoxygenase may be in the range of 0.4 / ⁇ g/mL to 28.8 ⁇ g/mL.
  • the reduction product may be 13-(9Z,l lE)-hydroxyoctadecadienoic acid, 13- (9E,llE)-hydroxyoctadecadienoic acid, 9-(10E,12Z)-hydroxyoctadecadienoic acid, or 9- (10E,12E)-hydroxyoctadecadienoic acid.
  • TPP triphenylphosphine
  • (9Z,l lE)-hydroxyoctadecadienoic acid, 8 to 12 molecules of 13 -(9E, HE)- hydroxyoctadecadienoic acid and 9 to 13 molecules of 9-(1OE, 12Z)- hydroxyoctadecadienoic acid may be respectively produced relative to 10 molecules of 9- (1 OE, 12E)-hydroxyoctadecadienoic acid.
  • the reducing agent is NaBH 4
  • 1:5 to 20 molecules of 13-(9Z 5 I lE)- hydroxyoctadecadienoic acid 9 to 13 molecules of 13-(9E,l lE)-hydroxyoctadecadienoic acid and 14 to 18 molecules of 9-(10E,12Z)-hydroxyoctadecadienoic acid may be respectively produced relative to 10 molecules of 9-(10E,12E)-hydroxyoctadecadienoic acid.
  • the hydroperoxidation may be carried out at a pH of 6 to 8 and a temperature of 20 to 30 ° C for 10 to 20 min.
  • the method may further comprise adding an organic solvent to elute the reaction product after completion of the hydroperoxidation.
  • the organic solvent may be at least one selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and any combination thereof.
  • Separation of the reduction product may be carried out by HPLC.
  • the present invention enables production of hydroxylinoleic acid with controlled distribution of various hydroperoxylinoleic acids from linoleic acid, via the use of maize lipoxygenase which simultaneously catalyzes 9- hydroperoxidation and 13-hydroperoxidation. Therefore, it is possible to provide various options of biosynthetic pathways that can cope effectively with a variety of stress applied to plants.
  • FIG. 1 illustrates reaction products from hydroperoxidation of linoleic acid using maize lipoxygenase
  • FIG. 2 illustrates SDS-PAGE patterns of lipoxygenase isolated and purified from E. coli BL21(DE3)pLysS/pRSETB/LOXl cells which expressed maize lipoxygenase and were subjected to sonic disruption
  • FIG. 3 illustrates the time course UV -vis spectra of a process where maize lipoxygenase produces hydroperoxylinoleic acid using linoleic acid as a substrate;
  • FIG. 4 illustrates Straight-Phase HPLC (SP-HPLC) analysis results for distribution of the reduction product obtained in Example 4 (A) and distribution of the reduction product obtained in Example 5 (B);
  • FIG. 5 illustrates the mass analysis spectra of trimethylsilylated 13 -(9Z, HE)- hydroxyoctadecadienoic acid prepared in Example 4;
  • FIG. 6 illustrates the mass analysis spectra of trimethylsilylated 9-(1OE, 12Z)- hydroxyoctadecadienoic acid prepared in Example 4. [Best Mode]
  • the present invention enables production of hydroxylinoleic acid with controlled distribution of various hydroperoxylinoleic acids by isolating maize lipoxygenase capable of simultaneously catalyzing 9-hydroperoxidation and 13-hydroperoxidation, preparing various hydroperoxylinoleic acids from linoleic acid using the maize lipoxygenase, reducing the resulting reaction products with a reducing agent, and then confirming distribution of the reduction products to thereby achieve controlled distribution of the reaction intermediates, hydroperoxylinoleic acids. Therefore, the present invention provides a basis which is capable of adopting diverse biosynthetic pathways that can cope actively with a variety of stress imposed on plants.
  • the present invention By controlling concentrations of linoleic acid and maize lipoxygenase according to the present invention, it is possible to easily modulate the distribution of various hydroperoxylinoleic acids which can be used as a starting material of diverse oxylipin biosynthetic pathways of plants. Further, the present invention is characterized in that it is possible to control a distribution ratio of hydroxylinoleic acid products depending upon kinds of reducing agents which reduces each of hydroperoxylinoleic acids.
  • the present invention provides a method for preparing hydroxylinoleic acid using maize lipoxygenase.
  • dual regiospecific hydroperoxidation of linoleic acid is first carried out using linoleic acid and maize lipoxygenase.
  • the oxidation of linoleic acid into hydroperoxylinoleic acid is catalytically accelerated.
  • LOXl lipoxygenase 1
  • Maize (Zea mays) lipoxygenase 1 (LOXl) is used as a catalyst that facilitates to induce dual positional specific or regiospecific hydroperoxidation of linoleic acid. That is, LOXl produces a variety of reaction products via cocatalysis of 9-hydroperoxidation and 13 -hydroperoxidation.
  • Zea mays LOXl is a protein having a molecular weight of about 97 kD in the natural state and an amino acid sequence as set forth in SEQ ID NO: 2.
  • the LOXl enzyme may be a protein having an amino acid sequence as set forth in SEQ ID NO: 2 or a protein having a sequence homology of more than 70% to the protein having an amino acid sequence of SEQ ID NO: 2. That is, the lipoxygenase protein in accordance with the present invention may encompass a protein having an amino acid sequence of SEQ ID NO: 2 isolated from maize (Zea mays) and a functional equivalent thereof.
  • the term "functional equivalent" refers to a protein that contains additions, substitutions and/or deletions of amino acid(s) and therefore has a sequence homology of at least more than 70%, preferably more than 80%, further preferably more than 90%, and still further preferably more than 95% to the amino acid sequence of SEQ
  • substantially the same physiological activity refers to an activity that enhances the plant stress resistance when the gene of interest is overexpressed in plants.
  • the hydroperoxidation is made by mixing and reacting linoleic acid and lipoxygenase.
  • Tri-Cl buffer may be used as a solvent for mixing of linoleic acid with lipoxygenase.
  • the mixed solution may contain a detergent to enhance the compatibility between hydrophilic and hydrophobic groups.
  • the detergent there is no particular limit to the detergent, as long as it is conventionally used in the art.
  • the hydroperoxidation may be carried out at a pH of 6 to 8 and room temperature of 20 to 30 ° C for 10 to 20 min.
  • the hydroperoxidation product will contain a hydroperoxy (-OOH) group at the C-9 or C-13 position of linoleic acid.
  • FIG. 1 illustrates reaction products from hydroperoxidation of linoleic acid using maize lipoxygenase.
  • lipoxygenases 13-LOX1 and 9- LOXl
  • LA linoleic acid
  • LA linoleic acid
  • HPODE hydroperoxyoctadecadienoic acid
  • 9-(1OE, 12Z)- hydroperoxyoctadecadienoic acid 9-(10E,12E)-hydroperoxyoctadecadienoic acid.
  • a distribution ratio of each reaction product may be controlled by a concentration of linoleic acid used as a substrate in the present invention or a concentration of lipoxygenase that catalyzes the dual positional specific hydroperoxidation of linoleic acid.
  • a concentration of linoleic acid may be controlled in the range of 0.03 mM to 0.5 mM to modify a distribution ratio of each reaction product.
  • a concentration of lipoxygenase may be controlled in the range of 0.4 //g/mL to 28.8 ⁇ g/mL to modify a distribution ratio of each reaction product.
  • reduction of the resulting hydroperoxidation product is carried out using triphenylphosphine (TPP) or NaBH 4 as a reducing agent.
  • the present invention features use of TPP or NaBH 4 as a reducing agent for reduction of hydroperoxidation products.
  • the method of the present invention may further comprise elution of the reaction products after completion of the hydroperoxidation. Elution of the reaction products may be carried out using any conventional method known in the art. For example, elution of the reaction products may be carried out using at least one organic solvent selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and any combination thereof.
  • the reduction product obtained from reduction of hydroperoxidation products using TPP or NaBH 4 as a reducing agent may be 13-(9Z,l lE)-hydroxyoctadecadienoic acid (HODE), 13-(9E,l lE)-hydroxyoctadecadienoic acid, 9-(10E,12Z)- hydroxyoctadecadienoic acid or 9-(10E,12E)-hydroxyoctadecadienoic acid.
  • HODE 13-(9Z,l lE)-hydroxyoctadecadienoic acid
  • 9-(10E,12Z)- hydroxyoctadecadienoic acid 9-(10E,12E)-hydroxyoctadecadienoic acid.
  • the present invention is characterized in that it is possible to control an amount of respective reduction products which are produced from reduction of hydroperoxidation products using the above-mentioned reducing agent, i.e. distribution of the reduction products.
  • the reducing agent is TPP
  • 11 to 15 molecules of 13-(9Z 5 I lE)- hydroxyoctadecadienoic acid, 8 to 12 molecules of 13-(9E,l lE)-hydroxyoctadecadienoic acid and 9 to 13 molecules of 9-(10E,12Z)-hydroxyoctadecadienoic acid may be respectively produced relative to 10 molecules of 9-(10E,12E)-hydroxyoctadecadienoic acid.
  • the reducing agent is NaBH 4
  • 15 to 20 molecules of 13-(9Z 5 I lE)- hydroxyoctadecadienoic acid 9 to 13 molecules of 13-(9E,l lE)-hydroxyoctadecadienoic acid and 14 to 18 molecules of 9-(10E,12Z)-hydroxyoctadecadienoic acid may be respectively produced relative to 10 molecules of 9-(10E,12E)-hydroxyoctadecadienoic acid.
  • the reduction of hydroperoxidation products may be carried out at a pH of 3 to 4 for 20 to 40 min.
  • Separation of the reduction product may be carried out by High Performance Liquid Chromatography (HPLC).
  • HPLC High Performance Liquid Chromatography
  • elution buffer for HPLC, as long as it is a nonsolvent conventionally used in the art.
  • a mixed solvent of hexane, alcohol and acetic acid may be used.
  • Example 1
  • E. coli strain harborin g a maize lipoxygenase gene (pX Ll /LOXl) introduced therein was cultured at 37 "C and the lipoxygenase gene (SEQ ID NO: 1, Genbank Accession No. AF271894) was then obtained using a plasmid DNA purification kit (QIAGEN Co., USA).
  • PCR Polymerase Chain Reaction
  • a primer pair for amplification of the gene of interest i.e., one primer of 5'- TGCAGCTGGTC ⁇ TATGGTCG-3' (SEQ ID NO: 3) having an Ndel restriction site and another primer of 5'-AAGATTCGAATTCAGCTCAG-S' (SEQ ID NO: 4) having an EcoRl restriction site.
  • the PCR product was introduced into a pGEM-T/Easy vector for amplification and then subcloned into NdeVEcoRl sites of a pRSETB vector to construct a recombinant LOXl expression vector, designated pRSETB/LOXl.
  • the recombinant vector pRSETB/LOXl was transfected into a BL21(DE3)pLysS strain which was then inoculated into an LB medium containing ampicillin (50 ⁇ g/mL) and chloramphenicol (35 mg/mL), followed by cultivation at 37 ° C for 16 hours.
  • the cells were inoculated at a dilution ratio of 1/100 in a fresh LB medium, and grown at 37 ° C to an optical density of 0.6 at 600 nm and then further cultured at 25 ° C for 12 hours after addition of an expression inducer isopropyl- ⁇ -D-thiogalactoside (IPTG) at a final concentration of 1 mM to thereby induce mass expression of the recombinant lipoxygenase
  • IPTG isopropyl- ⁇ -D-thiogalactoside
  • Bacterial cells were harvested by centrifugation of the cell culture (3,000 ⁇ g, 4 ° C and 15 min) and washed with 50 mM Tris-Cl buffer (pH 7.2). Then, the cells were centrifuged at 13,000*g and 4 0 C for another 1 hour and resuspended in 3 mL of Tris-Cl buffer (50 mM, pH 7.2) containing 0.1% Tween 20 and 0.2 mM protease inhibitor (phenylmethylsulfonyl fluoride (PMSF), Sigma), followed by sonic disruption to extract unpurified lipoxygenase.
  • Tris-Cl buffer 50 mM, pH 7.2
  • protease inhibitor phenylmethylsulfonyl fluoride (PMSF), Sigma
  • the obtained lipoxygenase-containing crude product was subjected to anion exchange chromatography to thereby purify lipoxygenase.
  • the crude product was passed through a column (Q-sepharose resin) equilibrated with 50 rnM Tris-Cl (pH 7.2) and then Tris-Cl (50 mM, pH 7.2) containing 50 mM NaCl was passed through the column.
  • Lane M represents a molecular weight marker
  • Lane 1 represents an unpurified extract obtained from the E. coli strain which was disrupted by sonication
  • Lane 2 represents a supernatant obtained by centrifugation of the unpurified crude extract
  • Lane 3 represents a precipitate obtained by centrifugation of the unpurified crude extract
  • Lane 4 represents lipoxygenase (97kD) purified by anion exchange chromatography. From the results of FIG. 2, it was confirmed that a staining marker of Lane 4 is lipoxygenase isolated from maize.
  • Example 4 Analysis of reduction products obtained from reduction of hydroperoxylinoleic acid using TPP
  • Hydroperoxylinoleic acid produced from linoleic acid using maize lipoxygenase, was reduced using TPP or NaBH 4 as a reducing agent and subjected to SP-HPLC to thereby separate four different regiospecific reduction products. Then, distribution of individual reduction products was examined.
  • FIG. 4A illustrates SP-HPLC analysis results for distribution of individual reduction products.
  • GC column HP-5MS with a size of 30 m x 0.25 mm x 0.25 ⁇ m was used. Injector and detector temperatures were set at 260 ° C and 300 °C , respectively. The column temperature was 100 ° C to 160 ° C (20 ° C /min), and 260 ° C to 280 ° C (4 ° C /min).
  • FIG. 5 illustrates mass analysis spectra of trimethylsilylated 13-(9Z,l lE)-hydroxyoctadecadienoic acid (HODE) corresponding to product I. Further, products III and IV had identical mass patterns.
  • FIG. 6 illustrates mass analysis spectra of trimethylsilylated 9-(10E,12Z)-hydroxyoctadecadie ⁇ oic acid (HODE) corresponding to product III.
  • Example 4 Referring to FIGS. 4, 5 and 6 in conjunction with Table 1, it was confirmed that four reduction products prepared in Example 4 correspond to I: 13-(9Z 5 I lE)-HODE, II: 13-(9E 5 I lE)-HODE, III: 9-(1OE, 12Z)-HODE, and IV: 9-(10E 5 12E)-HODE 5 respectively.
  • Example 5 Analysis of reduction products obtained from reduction of hydroperoxylinoleic acid using NaBH 4
  • Tris-Cl buffer 50 mM, pH 7.2 containing 0.5 mM linoleic acid and 0.05% Tween 20 (Uniqema) at 25 ° C for 15 min.
  • the reaction was stopped with addition Of NaBH 4 .
  • the resulting reaction solution was adjusted to have a pH of 3 by addition of IN HCl, and the pH-adjusted reaction solution was immediately passed through a solid-phase extraction cartridge (Sep-pak C 18 ) to afford a reduction product.
  • the reduction product was subjected to fractionation and mass analysis in the same manner as in Example 4.
  • Four fractionated reduction products obtained were designated I, II, III, and IV, respectively.
  • FIG. 4B illustrates SP-HPLC analysis results for distribution of individual reduction products.
  • Table 2 below shows SP-HPLC analysis results for distribution ratios of the reduction products of Examples 4 and 5 shown in FIG. 4. Referring to Table 2, it was confirmed that the reduction products produced according to Examples 4 and 5 exhibit varying distribution ratios depending upon kinds of reducing agents.
  • Example 6 Effects of linoleic acid concentrations on distribution of reduction products
  • Reduction products were obtained in the same manner as in Example 4, except that linoleic acid used as a substrate was added at various different concentrations of 0.03, 0.04, 0.05, 0.1 and 0.5 mM. Then, the products were fractionated, followed by mass analysis.
  • Table 3 shows distribution ratios of the reduction products with respect to varying concentrations of linoleic acid.
  • Reduction products were obtained in the same manner as in Example 4, except that lipoxygenase was added at a concentration of 1.0 ⁇ g or 72 ⁇ g. Then, the products were fractionated, followed by mass analysis.
  • Table 4 shows distribution ratios of the reduction products with respect to varying concentrations of lipoxygenase.
  • the present invention enables control of distribution of various hydroperoxylinoleic acids which can be utilized as a starting material of diverse oxylipin biosynthetic pathways of plants. Therefore, the present invention provides a basis which allows for broad options of biosynthetic pathways that can cope effectively with a variety of stress applied to plants.

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Abstract

La présente invention concerne, d'une part un procédé d'élaboration d'acide hydroxy-linoléique au moyen de lipoxygénase de maïs (Zea mays), et d'autre part l'acide hydroxy-linoléique ainsi obtenu. Le procédé de l'invention consiste d'abord à faire subir à l'acide linoléique une double hydro-peroxydation régiospécifique avec une solution mixte d'acide linoléique et de lipoxygénase du maïs; puis à réduire à la triphényl-phosphine ou au NaBH4 le produit de réaction obtenu; et enfin à séparer le produit de réduction. Le procédé de la présente invention permet une production d'acide hydroxy-linoléique tout en gérant la répartition des divers acides hydro-peroxy-linoléiques provenant de l'acide linoléique, grâce à l'utilisation de la lipoxygénase du maïs capable de catalyser en même temps la 9-hydro-peroxydation et la 12-hydro-peroxydation. L'invention permet ainsi de proposer plusieurs canaux de biosynthèse donnant la possibilité de traiter divers stress subis par les végétaux.
PCT/KR2008/001049 2007-04-30 2008-02-22 Procédé d'élaboration d'acide hydroxy-linoléique au moyen de lipoxygénase de maïs et acide hydroxy-linoléique ainsi obtenu WO2008133398A1 (fr)

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KR20070041982 2007-04-30
KR10-2007-0041982 2007-04-30
KR1020080012663A KR100938714B1 (ko) 2007-04-30 2008-02-12 옥수수 유래 리폭시지나아제 효소를 이용한히드록시리놀레산의 제조방법 및 이에 의해 제조된히드록시리놀레산
KR10-2008-0012663 2008-02-12

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5942661A (en) * 1996-01-19 1999-08-24 The Texas A&M University System Method of inhibiting mycotoxin production in seed crops by modifying lipoxygenase pathway genes
US6271018B1 (en) * 2000-03-29 2001-08-07 Vanderbilt University Muskmelon (Cucumis melo) hydroperoxide lyase and uses thereof
KR20010085101A (ko) * 2001-08-08 2001-09-07 한옥수 광범위 기질 특이적 리폭시지나아제 및 이의 용도
KR20030018167A (ko) * 2001-08-27 2003-03-06 김태윤 불포화 지방산 리폭시지네이즈 대사체의 대량 정제 방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5942661A (en) * 1996-01-19 1999-08-24 The Texas A&M University System Method of inhibiting mycotoxin production in seed crops by modifying lipoxygenase pathway genes
US6271018B1 (en) * 2000-03-29 2001-08-07 Vanderbilt University Muskmelon (Cucumis melo) hydroperoxide lyase and uses thereof
KR20010085101A (ko) * 2001-08-08 2001-09-07 한옥수 광범위 기질 특이적 리폭시지나아제 및 이의 용도
KR20030018167A (ko) * 2001-08-27 2003-03-06 김태윤 불포화 지방산 리폭시지네이즈 대사체의 대량 정제 방법

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