[go: up one dir, main page]

WO2000008168A1 - Ameliorations des souches de streptomyces grace a des genes impliques dans la biosynthese de la tylosine - Google Patents

Ameliorations des souches de streptomyces grace a des genes impliques dans la biosynthese de la tylosine Download PDF

Info

Publication number
WO2000008168A1
WO2000008168A1 PCT/ES1999/000248 ES9900248W WO0008168A1 WO 2000008168 A1 WO2000008168 A1 WO 2000008168A1 ES 9900248 W ES9900248 W ES 9900248W WO 0008168 A1 WO0008168 A1 WO 0008168A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
gene sequence
encoded product
tylosin
fradiae
Prior art date
Application number
PCT/ES1999/000248
Other languages
English (en)
Spanish (es)
Inventor
Roberto Fouces Martinez
Encarnación MELLADO DURAN
Bruno Diez Garcia
Manuel Esteban Morales
Ermanno Bernasconi
José Luis BARREDO FUENTE
Original Assignee
Antibioticos, S.A.U.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Antibioticos, S.A.U. filed Critical Antibioticos, S.A.U.
Priority to AU52911/99A priority Critical patent/AU5291199A/en
Publication of WO2000008168A1 publication Critical patent/WO2000008168A1/fr

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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/001—Oxidoreductases (1.) acting on the CH-CH group of donors (1.3)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/36—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Actinomyces; from Streptomyces (G)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0077—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14) with a reduced iron-sulfur protein as one donor (1.14.15)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1003—Transferases (2.) transferring one-carbon groups (2.1)
    • C12N9/1007—Methyltransferases (general) (2.1.1.)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • 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.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • C12N9/1077—Pentosyltransferases (2.4.2)
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • 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
    • C12N9/90—Isomerases (5.)
    • 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
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/44—Preparation of O-glycosides, e.g. glucosides
    • C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
    • C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin

Definitions

  • the present invention relates to a method based on the use of genes of the tylosin biosynthetic cluster for increasing the production of tylosin in S. fradiae or for the production of new hybrid metabolites in related microorganisms ⁇ Streptomyces spp. ). It describes: (I) a method to isolate a DNA fragment of S. fradiae containing 11 genes, 10 of them belonging to the cluster of biosynthetic genes of tylosin and (II) the expression of said genes in the microorganism producing tylosin S. fradiae, as well as in other related microorganisms ⁇ Streptomyces spp. ) achieving an increase in tylosin production in the first case and the production of new hybrid antibiotics in the second.
  • Tylosin is a macrolide antibiotic composed of a 16-member macrocyclic lactone or ring to which 3 modified sugars bind: micarous, micaminous and microsane.
  • microorganisms capable of producing tylosin are the aforementioned S. fradiae (Seno, ET et ai., 1977. Antimicrob. Agents. Chemother. 21, 758), S. rimosus (Pape, H. and Brillinger, RH 1973. Arch. Microbiol. 88, 25-35) and S. hygroscopicus (Jensen, A. L. et al., 1964. In: Antimicrobial agents and chemotherapy-1963, JC Sylvester ed., American Society for Microbiology, Washington D. C). For the industrial production of tylosin S is used. fradiae.
  • Tylosin biosynthesis begins with the formation of the macrocyclic ring, by a process similar to that of fatty acid biosynthesis, but which differs from the latter in the great variety of isolated precursors that can be incorporated and in the partial or total absence of reactions that reduce the ⁇ -ketone group formed after each chain extension.
  • the macrocyclic ring is composed of 2 acetates, 5 propionates and a butyrate.
  • the enzymatic activity responsible for synthesizing it, called polyketide synthase is encoded by the tylG gene.
  • Said reactions are as follows: (I) addition of micaminose in the 5-OH position of the macrolide ring to form O-micamino-siltilactone; (II) oxidation of the methyl group in C-20 to form 20-dihydro-23-deoxy-O-mycaminosyllactone; (III) oxidation of the hydroxymethyl in C-20 to formyl group to produce 23-deoxy-O-micaminosiltilactone; (IV) oxidation of methyl group in C-23 to hydroxymethyl to form a 0-mycosylamino-methylonolide; (V) addition of 6-deoxyalose to the C-23 OH group of the macrolide ring to form demethyllactone; (VI) addition of micarose to 4'OH of the micaminose to form demethyl-macrocin; (VII) methylation of 2 '' OH of 6-deoxyalose to form macrocin and (VIII) methylation of 3 ''OH of 6-de
  • the technique called combinatorial biosynthesis, has allowed the biosynthesis, among other compounds, of erythromycin analogues and peptide antibiotics, of spiramycin and tylosin derivatives, as well as of numerous aromatic and reduced metabolites by manipulating genes that code for polyketide synthases (Madduri, K. et al. 1998. Nature Biotechnology 16: 69-74).
  • This new approach has been used for the production of isovaleryl spiramycin by Streptomyces ambofaciens strains in which the StEptomyces thermotolerans carE gene has been expressed, which codes for the deoxy hexose O-acyltransferase enzymatic activity.
  • This enzyme causes the conversion of the 4 '' -hydroxyl group of the spicamycin micarous residue into its isovaleryl ester (Epp, JK et al. 1989. Gene 85: 293-301).
  • the production of the antitumor compound 4 '-epidoxorubicin has been described in a strain of Streptomyces peucetius in which the dnmV gene has been inactivated and the avrE and / or eryBIV genes of Streptomyces avermi tilis and Saccharopolyspora erythraea respectively (Madduri) have been introduced , K. et al. 1998. Nature Biotechnology 16: 69-74).
  • tylosin biosynthetic genes described in the present invention into microorganisms producing macrolide or related antibiotics can be used for the production of new hybrid antibiotics.
  • the genes involved in the biosynthesis of the sugars included in the tylosin molecule could be introduced into microorganisms producing macrolide antibiotics to achieve biosynthesize hybrid antibiotics.
  • genes responsible for the myosin biosynthesis expressed in an erythromycin-producing microorganism will enable the production of erythromycin-derived mycinosil.
  • fradiae ATCC 19609 both as a source of deoxyribonucleic acid (hereinafter referred to as DNA) and ribonucleic acid (hereinafter referred to as RNA).
  • DNA deoxyribonucleic acid
  • RNA ribonucleic acid
  • fradiae ATCC 19609 a library was constructed in the lambda-GEM12 (Promega) phage vector according to standard procedures (Sambrook, J. et al. 1989. Molecular cloning, a laboratory manual, Cold Spring Harbor Laboratory Press).
  • Plasmid pALFIA Figure 2 was constructed by subcloning an 11.5 kb Sacl fragment into pBluescript I KS (+). Likewise, a PvuII fragment was purified, which included both the recombinant phage insert and a part belonging to the lambda-GEM12 vector arm.
  • Plasmid pALFIA was propagated in E. coli strain DH5 ⁇ in order to obtain, by means of the alkaline lysis procedure (Sambrook, J. et al. 1989. Molecular cloning, a laboratory manual, Cold Spring Harbor Laboratory Press), sufficient quantity of DNA for successive processes.
  • the location of the tylF gene was determined by hybridization by the Southern technique of the above-mentioned synthetic oligonucleotide (5 ' GCTCGATGTAGAGATCG 3') with digestions of the plasmid pALFIA with a series of endo- restriction nucleases.
  • the tylF gene was identified in a 5.7 kb BamHI fragment and in another 4.1 kb BglII-.Ba.7iHI.
  • a preliminary restriction map of the area was prepared ( Figure 2), which was used as the basis for the sequencing process.
  • the determination of the nucleotide sequence (example 1) of a fragment of 12,905 base pairs (hereinafter referred to as bp) showed the existence of 11 open reading frames (hereinafter referred to as ORF), of which 10 belong to the cluster tylosin biosynthetic.
  • ORF 11 open reading frames
  • Plasmids, pALF287 or pALFIB, which contain a series of tylosin biosynthetic genes, were introduced into the S. fradiae ATCC 19609 collection strain and into the tylosin S superproductive strains. fradiae 302 and S. fradiae 455, achieving a decrease in the accumulated macrocin levels and an increase in the level of tylosin production with respect to parental strains.
  • Plasmid pALF250 was also constructed ( Figure 5), which is formed by pULVK99 plus a DNA fragment of approximately 1.4 kb containing the trlB gene, a determinant of tylosin resistance of S. fradiae.
  • pALF250 was introduced by transformation into S host cells. lividans ATCC 1326 and S. parvulus DSM 40048, using the tylosin resistance marker as a selective method to maintain plasmid pALF250 autonomously in said host. The transformants of S.
  • Carrier lividans of said plasmid were selected using tylosin concentrations of 250 to 500 ⁇ g / ml, although some of them were capable of growing in concentrations of 3.5 mg / ml, the minimum inhibitory concentration of tylosin being for S. untransformed lividans of 200 ⁇ g / ml.
  • the transformants of S. Parvulus were able to grow in concentrations of 200 ⁇ g / ml of tylosin, the minimum inhibition concentration of tylosin being for S. untransformed parvulus of 100 ⁇ g / ml.
  • This new resistance marker is very useful, since the number of selectable genetic markers in Streptomyces spp. It is not very broad and the use of the tlrB gene expands that restricted field.
  • Plasmids pALF17 and pALF18 were subdivided into smaller fragments using the 2 Ba ⁇ nHI restriction sites and the unique Sacl site. The resulting fragments were subcloned into the pBluescript I KS (+) vector giving rise to the following plasmids: pALF71, pALF72, pALF73, pALF74, pALF76 and pALF77 ( Figure 2).
  • plasmid pALF32 was double digested with the enzymes BamHI and BglII, the fragments obtained (4.1 kb and 165 bp) were subcloned into pBluescript II KS (+).
  • the plasmids obtained were called pALF2 and pALFIO (4.1 kb fragment) and pALF21 (165 bp fragment) ( Figure 2).
  • pALFl ⁇ and pALF267 were obtained. The first covers the BamHI restriction site that separates the 4.1 kb BglI l-BamUl fragment and the 1 kb BamHI fragment and the second does the same on the BamHI site that separates the latter from the 2.1 kb BamHI fragment (figure 2).
  • ORFs correspond to the following genes: 0RF1 with the ddcA gene, ORF2 with the tlrB gene, ORF3 with the tylN gene, ORF4 with the tylE gene, ORF5 with the tylD gene, ORF6 with the tylH2 gene, ORF7 with the tylHl gene, ORF8 with the tylF gene, ORF9 with the tylO gene, ORF10 with the tylP gene and ORF11 with the tylQ gene.
  • the preparation of S protoplasts Fradiae was performed by inoculating 50 ml of TSB medium supplemented with 0.4% glycine and 5 mM MgCl 2 , in a 500 ml flask, with a spore suspension of the strain to be transformed. The microorganism was grown at 30 ° C and 250 rpm for 48 hours. After that time, the mycelium was collected by centrifugation at 3,000 rpm for 7 minutes at temperature Ambience in two Sorvall SS34 tubes. The sediment was washed with 25 ml of 10% sucrose, centrifuging under the above conditions.
  • the sediment from 25 ml of culture was resuspended in 5 ml of buffer P containing 1 mg / ml lysozyme and 0.4 mm diameter glass beads were added.
  • the rest of the culture can be frozen at -20 ° C, resuspended in 25 ml of 10% sucrose, to be used in subsequent transformations.
  • the mycelium resuspended in buffer P was incubated at 19 ° C by periodically shaking the tube to homogenize the mycelium with the glass balls. The incubation was prolonged until it was observed under the microscope that a drop of mycelium suspension was completely smooth by adding a drop of a 10% SDS solution.
  • the transformation process began by defrosting one of the aliquots of the protoplast suspension in a 37 ° C bath. Next, 1 ⁇ g of the DNA we wanted to introduce was added and mixed by gentle agitation of the tube, immediately adding 500 ⁇ l of 50% PEG dissolved in P buffer. Next, the contents of the tube were mixed by rapid inversion and the transformation mixture was incubated for 3-5 minutes at room temperature. Subsequently, 5 ml of soft R5 (R5 medium containing 0.6-1% agar) were plated, with the transformed protoplasts (protoplasts, DNA and PEG), in 1-2 Petri dishes of R5 medium.
  • the transforming strains obtained by the procedure described in Example 2 were fermented according to previously described protocols (Baltz RH and Seno ET, 1981, Antimicrobial Agents and Chemotherapy 20: 214-225; Cox KL et al. 1994. European Patent Specification 0238323B1), adding thiostreptone (antibiotic to which the plasmid confers resistance) at a concentration of 5 ⁇ g / ml.
  • the transformants corresponding to plasmids pALFIB (figures 2 and 3) and pALF287 (figures 2 and 4) showed significant increases (10-50% depending on the host strain) in tylosin production when compared with the parental strain. Also, because both plasmids include the tylF gene, the transformants showed a drastic decrease (25-50 times) in the accumulation of the macrocin precursor in fermentation broths.
  • FIGURE 1 S genome region. fradiae that includes the grouping of biosynthetic genes of tylosin.
  • FIGURE 2 Restriction map of the S. fradiae genome region that includes the clustering of tylosin biosynthetic genes.
  • the different constructed plasmids are shown at the bottom.
  • the insert corresponding to plasmids pALFIA and pALF2A consist of fragments of 11.5 kb Sacl and 16 kb PvuII respectively, while the insert of plasmid pALFIB corresponds to a fragment of 12.9 kb BglII-BamHI.
  • the clones that appear with arrowheads are those used in the sequencing process.
  • FIGURE 3 Restriction map of the 20.8 kb plasmid pALFIB, which includes the grouping of biosynthetic tylosin genes described in the present invention.
  • FIGURE 4 Restriction map of the 13.5 kb plasmid pALF287, which includes part of the tylosin biosynthetic gene cluster described in the present invention.
  • FIGURE 5 Restriction map of the 9.2 kb plasmid pALF250, which includes the tyrosine resistance gene t rB described in the present invention.

Landscapes

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

Abstract

L'invention concerne une amélioration des souches de Streptomyces grâce à des gènes impliqués dans la biosynthèse de la tylosine et un procédé consistant à utiliser des gènes contenus dans le groupe de biosynthèse de la tylosine pour l'augmentation de la production de la tylosine chez S. fradiae. Ce procédé consiste (I) à isoler un fragment d'ADN de S. fradiae contenant 11 gènes, dont 10 appartiennent au groupe de gènes impliqués dans la biosynthèse de la tylosine et (II) à exprimer ces gènes dans le micro-organisme S. fradiae producteur de la tylosine, ainsi que dans d'autres micro-organismes apparentés (Streptomyces spp.), assurant ainsi une augmentation de la production de tylosine dans le premier cas et la production de nouveaux antibiotiques hybrides dans le second cas.
PCT/ES1999/000248 1998-08-05 1999-08-02 Ameliorations des souches de streptomyces grace a des genes impliques dans la biosynthese de la tylosine WO2000008168A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU52911/99A AU5291199A (en) 1998-08-05 1999-08-02 Improvements to strains of streptomyces through the use of biosynthetic genes oftylosine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ESP9801682 1998-08-05
ES009801682A ES2155342B1 (es) 1998-08-05 1998-08-05 Mejoras de cepas de streptomyces mediante la utilizacion de genes biosinteticos de tilosina.

Publications (1)

Publication Number Publication Date
WO2000008168A1 true WO2000008168A1 (fr) 2000-02-17

Family

ID=8304796

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ES1999/000248 WO2000008168A1 (fr) 1998-08-05 1999-08-02 Ameliorations des souches de streptomyces grace a des genes impliques dans la biosynthese de la tylosine

Country Status (3)

Country Link
AU (1) AU5291199A (fr)
ES (1) ES2155342B1 (fr)
WO (1) WO2000008168A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1457558A4 (fr) * 2001-10-19 2005-10-12 Sumitomo Chemical Co Proteines du metabolisme permettant de lutter contre les mauvaises herbes, leurs genes et leur utilisation
US10336790B2 (en) * 2014-03-10 2019-07-02 Rensselaer Polytechnic Institute Anti-microbial peptides and method for designing novel anti-microbial peptides

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0364130A2 (fr) * 1988-09-29 1990-04-18 Eli Lilly And Company Nouveaux gènes de biosynthèse de la tylosine
US5672497A (en) * 1986-03-21 1997-09-30 Eli Lilly And Company Method for increasing the antibiotic-producing ability of antibiotic-producing microorganisms

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5672497A (en) * 1986-03-21 1997-09-30 Eli Lilly And Company Method for increasing the antibiotic-producing ability of antibiotic-producing microorganisms
EP0364130A2 (fr) * 1988-09-29 1990-04-18 Eli Lilly And Company Nouveaux gènes de biosynthèse de la tylosine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FISHMAN S.E. ET AL: "Cloning Genes for the Biosynthesis of a Macrolide Antibiotic", PROC. NATL. ACAD. SCI. USA,, vol. 84, 1987, pages 8248 - 8252, XP000887108, DOI: doi:10.1073/pnas.84.23.8248 *
FOUCES R. ET AL: "The Tylosin Biosynthetic Cluster from Streptomyces Fradiae: Genetic Organization of the Left Region", MICROBIOLOGY, vol. 145, 1999, pages 855 - 868 *
GANDECHA A.R. ET AL: "Analysis of Four Tylosin Bio-synthetic Genes from the tyILM Region of the Streptomyces Fradial Genome", GENE, vol. 184, 1997, pages 197 - 203 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1457558A4 (fr) * 2001-10-19 2005-10-12 Sumitomo Chemical Co Proteines du metabolisme permettant de lutter contre les mauvaises herbes, leurs genes et leur utilisation
US7745699B2 (en) 2001-10-19 2010-06-29 Sumitomo Chemical Company, Limited Weed controller metabolism proteins, genes thereof and use of the same
US8293979B2 (en) 2001-10-19 2012-10-23 Sumitomo Chemical Company, Limited Weed controller metabolism proteins, genes, thereof and use of the same
US10336790B2 (en) * 2014-03-10 2019-07-02 Rensselaer Polytechnic Institute Anti-microbial peptides and method for designing novel anti-microbial peptides
US11202449B2 (en) 2014-03-10 2021-12-21 Rensselaer Polytechnic Institute Anti-microbial peptides and method for designing novel anti-microbial peptides

Also Published As

Publication number Publication date
AU5291199A (en) 2000-02-28
ES2155342A1 (es) 2001-05-01
ES2155342B1 (es) 2001-12-01

Similar Documents

Publication Publication Date Title
US5824513A (en) Recombinant DNA method for producing erythromycin analogs
Stanzak et al. Cloning and expression in Streptomyces lividans of clustered erythromycin biosynthesis genes from Streptomyces erythreus
Ruan et al. Acyltransferase domain substitutions in erythromycin polyketide synthase yield novel erythromycin derivatives
EP2271666B1 (fr) Groupe de gènes nrps-pks, sa manipulation et son utilité
US6251636B1 (en) Recombinant oleandolide polyketide synthase
Stutzman-Engwall et al. Multigene families for anthracycline antibiotic production in Streptomyces peucetius.
EP0238892A2 (fr) Vecteurs de clonage pour streptomyces et leur utilisation pour la production des antibiotiques macrolides
WO1993013663A1 (fr) Procede permettant d'effectuer la biosynthese de polycetides specifiques
ES2812256T3 (es) Agrupación de genes de biosíntesis de carrimicina
JPH04346786A (ja) ストレプトミセス・アウレオフアシエンスからのクロロテトラサイクリンおよびテトラサイクリンの形成の生合成経路のクローニングおよびストレプトミセス・リビダンス中のその発現
JP2005529579A (ja) ポリケチドの生合成のための過剰産生宿主
CN101363022A (zh) 替曲卡星a的生物合成基因簇及其应用
WO2000008168A1 (fr) Ameliorations des souches de streptomyces grace a des genes impliques dans la biosynthese de la tylosine
ES2346412T3 (es) Genes biosinteticos para la produccion de insecticidas de tipo butenilespinosina.
EP1383783A2 (fr) Agregat de genes de la biosynthese de leinamycine et ses composantes et leurs utilisations
CN101892185B (zh) 产表柔红霉素的天蓝淡红链霉菌基因工程菌及其制备方法
CN101892186B (zh) 产表柔红霉素的天蓝淡红链霉菌的基因工程菌及其制备方法
JP2005511054A (ja) ハイブリッドグリコシル化産物及びそれらの産生及び使用
CN106916834B (zh) 化合物的生物合成基因簇及其应用
JPS63313589A (ja) マイクロモノスポラのための遺伝子系
EP1183369A1 (fr) Polycetides et leur synthese
JP4633519B2 (ja) ミデカマイシン高生産菌
ES2220938T3 (es) Proceso para producir antraciclinas y sus intermedios.
EP0806480A2 (fr) Groupe de gènes pour la production de frénolicine
CN102021187A (zh) Fr-008聚酮抗生素生物合成相关的基因簇

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA