[go: up one dir, main page]

WO2006014047A1 - Enzyme impliquee dans la biosynthese de la ribostamycine et les genes correspondants - Google Patents

Enzyme impliquee dans la biosynthese de la ribostamycine et les genes correspondants Download PDF

Info

Publication number
WO2006014047A1
WO2006014047A1 PCT/KR2005/001801 KR2005001801W WO2006014047A1 WO 2006014047 A1 WO2006014047 A1 WO 2006014047A1 KR 2005001801 W KR2005001801 W KR 2005001801W WO 2006014047 A1 WO2006014047 A1 WO 2006014047A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
encoding
amino acid
acid sequence
rbm
Prior art date
Application number
PCT/KR2005/001801
Other languages
English (en)
Inventor
Jin Suk Woo
Jae Kyung Sohng
Hei Chan Lee
Kwang Kyoung Liou
Young Soo Jung
Sun Youp Kang
Dae Hee Kim
Wun Min Seo
Ji Young Yang
Original Assignee
Genechem Inc.
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 Genechem Inc. filed Critical Genechem Inc.
Publication of WO2006014047A1 publication Critical patent/WO2006014047A1/fr

Links

Classifications

    • 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
    • C12N9/10—Transferases (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
    • C12N9/10—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
    • 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
    • C12N9/88—Lyases (4.)

Definitions

  • DOS-containing aminoglycoside-aminocyclitol (AmAc) antibiotics are clinically important antibiotics since they have broad antibacterial spectrum activity.
  • these antibiotics can be classified into two major types: one with 4, 5-disubstituted deoxystreptamine (DOS), such as ribostamycin (Rbm), butirosin (Bn) and neomycin (Nm), and the other with 4, 6- disubstituted DOS, such as kanamycin (Km) and gentamicin (Gm).
  • DOS 5-disubstituted deoxystreptamine
  • Rbm ribostamycin
  • Bn butirosin
  • Nm neomycin
  • 6- disubstituted DOS such as kanamycin (Km) and gentamicin (Gm).
  • Rbm is composed of three subunits: DOS, neosamine C and ribose. It is different from Bn B in that the Cl-NH 2 of the latter antibiotic is conjugated with (2R)-4- amino-2 -hydroxy butyric acid (AHBA) (FIG 1). The conjugation of this side chain has revived the clinical utility of Km B as arbekacin (ABK), and KmA as amikacin.
  • the present inventors have cloned the gene fragment expected as Rbm biosynthetic gene cluster in chromosome DNA of S.ribosidificus to sequence and had Rbm biosynthetic genes expressed at a large amount and thus confirmed that said cloned gene fragment is Rbm biosynthetic gene cluster by biochemical analysis of purified proteins, thereby completing the present invention.
  • An object of the present invention is to provide DNA sequences of gene cluster participating in Rbrn biosynthesis.
  • Still another object of the present invention is to provide aminoglycoside-3-N- acetyltransferase[AAC(3)] as Rbm biosynthetic enzyme and genes encoding the enzyme.
  • the ribostamycin(Rbm) biosynthetic gene cluster is derived from Streptomyces ribosidificus.
  • the present invention provides a recombinant vector containing the genes, bacteria transformed with the recombinant vector, and a method for preparing DOI synthase, in which the transformed bacteria are cultured.
  • the present invention provides aminoglycoside-3-N- acetyltransferase[AAC(3)] which is a ribostamycin(Rbm) biosynthetic enzyme with an amino acid sequence of SEQ ID NO: 4 and genes(rhml) encoding the AAC(3) .
  • FIG. 1 shows the structures of aminoglycoside-aminocyclitols (AmAcs): A., butirosin B(Bn B) and B., ribostamycin.
  • AmAcs aminoglycoside-aminocyclitols
  • FIG 2 is a gene map of butirosin and ribostamycin biosynthetic gene clusters. The genes encoding similar proteins are indicated by the similar arrows. B, E and H in the map stand for BamHl, EcoRl and Hind ⁇ l, respectively.
  • FIG 4 shows SDS-PAGE analysis of RbmA expressed for 3 days in S. lividans TK-24.
  • Lane 1 represents the whole proteins expressed in S. lividans TK24/pIBR25 as a control
  • lane 2 represents the whole proteins of S. /m ⁇ m.s i TK24/pBS2 J and M represents protein marker(Novagen, USA).
  • a box represents RbmA.
  • FIG 5 shows the result of HPLC analysis of DOI formation using the cell-free extract of S. lividans transformant.
  • the arrow indicates the peak for the oxime derivative of DOI.
  • FIG. 7 is a photograph of TLC analysis on whether AmAcs are acetylated or not after a reaction with Rbml.
  • Lanes 1, 2, 3, 4, 5 and 6 represent samples of a control group without adding coenzyme A to kanamycin, ribostamycin, apramycin, neomycin, gentamicin, and spectinomycin, and 1' to 6' are corresponding acetylated samples by adding coenzyme A, respectively.
  • FIG. 8 represents the result of antibacterial assay of acetylated AmAcs.
  • Apm, Km, Rbm, Nem, and Gm represent apramycin, kanamycin, ribostamycin, neomycin and gentamicin and Rxn, Ref and Std represent reaction, reference and standard samples, respectively.
  • S. ribosidificus(ATCC2l294) and S. lividans TK24(US20030142745 Al) were grown in ISP2(Difco, USA) and R2YE(US 5,843,735) media in liquid or on agar plates at 28°C.
  • Echerichia coli XLl blue(Stratagene, USA) and E. coli BL21 (DE3) (Stratagene, USA) were used as a host for sub-cloning and a host for gene expression, respectively.
  • pOJ446(US 20040053274A1) was used for the construction of the genomic library, whereas pET-32a(+) (Novagen, USA) was used for expressing genes in E. coli.
  • pIBR25(Stha ⁇ it, B. et al., FEBS Lett., 566:201, 2004) was used for the expression of gene in S. lividans TK24.
  • thiostrepton 50 ⁇ g ml "1 ) was supplemented in the medium.
  • Example 1 The construction of the DNA library and screening of the cosmid library
  • the genomic DNA of S. ribosidificus was partially digested with diluted Mbol over various time intervals (0.5 to 5 min), and aliquots were analyzed by agarose gel electrophoresis.
  • the cleaved sample containing 35 ⁇ 45 kb fragments was pooled, treated with alkaline phosphatase and then ligated to pOJ446 digested with BamHI and Hpal.
  • Packaging of the ligated sample was carried out in vitro using Gigapack III XL packaging extract (Stratagene, USA).
  • a cosmid library of S. ribosidificus was screened with two different probes. Partial sequences of DOI synthase and GLA obtained from the genomic DNA of S. ribosidificus were used as the probes for the screening of the cosmid library.
  • DOI synthase probe was labeled with 32 P-dCTP (Perkin-Elmer Life Sciences, USA) using a random primer labeling kit (Stratagene, USA), purified by gel filtration, and used for screening.
  • Hybridization was carried out for 6 hr with each probe at 65°C in 10 ml of 2 x SSC. Sequencing was carried out in an automatic sequencer by the dideoxy chain termination method and the sequences were assembled using the DNA Star program package (DNASTAR, Inc., USA). Potential ORFs(open reading frames) were identified using the FramePlot(http://www.nih.go.jp/ ⁇ jun/cgi- bin/frameplotpl) and they were searched for homologies using BLAST(http://www.ncbi.nlm.nih.gov/BLAST/).
  • Colony hybridization of the cosmid library of S. ribosidificus with core nucleotide sequences of DOI synthase revealed four independent clones. Expected size of PCR products, 345 bp and 270 bp, were obtained from the cosmids using primers of SEQ ID NO: 27 to SEQ ID NO: 30 designed from the core sequences of DOI synthase and GLA respectively. An insertion of over 32 kb fragment was designated as pRBM4 and sequenced.
  • a cluster of 26 ORFs was determined in 31.892 kb region of pRBM4 (FIG. 2.).
  • This cluster includes putative Rbm biosynthetic genes (rbmA, rbmB, rbmC, rbniD, rbmG and rbmH), resistance genes (rbml and rph ) and transport genes (rbmE and rbmF).
  • the predicted product of rbmA was 62%, 55% and 37% identical to TbmA (accession NO: CAE22471), GtmA (accession NO: BAC41210), and BtrC (accession NO: 41210) from the Tbm-producer S. tenrebrarius, Gm-producer M. echinospora and Bn-producer B. circulans, respectively. All of these proteins catalyze the formation of DOI utilizing G-6-P in the presence of NAD + and Co 2+ (FIG. 3.).
  • RbmG represents another TacD homologue in Rbm cluster.
  • RbmH an aminotransferase
  • rbml and rph flank the Rbm biosynthetic genes cluster.
  • the product of the former gene, rbml is homologous to the AAC (3) from various AmAc producers: 70% to AACC8 (accession NO: P29809) from S.fradiae, 58% to AACC7(accession NO: P30180) from S. rimosus and 55% to kan (accession NO: BAA78619) from Streptomyces griseus, respectively.
  • rph encodes a protein which confers resistance on the host by phosphorylating Rbm in S. ribosidificus( ⁇ oshiko, S. et al., Gene, 68:285, 1988).
  • Several other hypothetical proteins identified in the Rbm cluster are summarized in the Table 1.
  • Table 1 Summary of proteins identified in Rbm biosynthetic gene cluster
  • cinnamoneus 0 Hypothetical protein racB 178 N CAD60535 (51) S. cinnamoneus 0 Hypothetical protein racC 378 N CAD60536 (34) S. cinnamoneus 0 Hypothetical protein rph 265 N AAC32025 (99) S. ⁇ bosidiflcus" Aminoglycoside phosphotransferase racD 125 N - - Hypothetical protein racE 270 N - - Hypothetical protein racF 258 N - - Hypothetical Protein racG 199 N AAP92498 (85) S. vinaceus” Type II thioesterase racH 300 BtrP (43) BAC41220 (43) B. circulans.
  • plasmids were constructed to produce DOI synthase (RbmA) fused with thioredoxin and histidine (18 kDa) at the amino-terminus.
  • the primers SEQ ID NO: 31 and 32 were used to amplify rbmA from ⁇ RBM4.
  • the amplified fragments were cloned into BamHl- and Hm ⁇ II-digested pET- 32a(+)(Novagen, USA) and pIBR25 to form pBSl and pBS2 respectively.
  • the recombinant pDOI-2 for the expression o ⁇ btrC (1.1 kb) was constructed by general method(Kharel, M.K. et al., FEMS Microbiol. Lett., 230:185, 2004).
  • rbml was amplified with the primers SEQ ID NOs: 33 and 34 and the product was cloned into B ⁇ mHl- and Hm ⁇ II-digested pET- 32a(+) to form ⁇ BS3.
  • E. coli BL21 (DE3)/pBSl was grown in 10 ml LB medium containing ampicillin at 37°C and 250 rpm for 8 hr. The culture was then transferred to 100 ml of LB medium and incubated at 37°C. When absorbancy is 0.6 at OD 60O , IPTG was added to a final concentration of 0.4 mM and the incubation was continued at 25°C for 20 hr. The culture was harvested by centrifugation at 6000 x g for 10 min, washed with Tris-HCI buffer (50 mM, Tris- HCI, pH7.5 and 0.2 mM Co 2+ ) and stored at -20°C for 6 hr.
  • Tris-HCI buffer 50 mM, Tris- HCI, pH7.5 and 0.2 mM Co 2+
  • Futhermore, S. lividans TK24/pBS2 was cultured in 25 ml R2YE medium supplemented with thiostrepton(50 ⁇ g ml "1 ) for 3 days and the culture (5 ml) was inoculated to 250 ml of R2YE medium and continued the incubation under identical conditions for 5 days.
  • the samples (50 ml) harvested at various times (24 hr, 48 hr, 72 hr, 96 hr and 120 hr) were washed with Tris-HCI buffer (50 mM Tris- HCI, ⁇ H7.5, and 0.2 mM Co 2+ ) and stored at -20 0 C for 6 hr.
  • E. coli BL21 (DE3)/pBS3 was grown by the same method as that for E.coli BL21 (DE3)/pBSl to induce the expression using IPTG. The incubation was continued at 20 0 C for 12 hr.
  • the cell pellets prepared in the example 4 were thawed, suspended in the Tris-HCI buffer and sonicated over various times. The disrupted cells were centrifuged at 12000 x g for 20 min to remove supernatant. Rbml was purified by Ni 2+ -affinity chromatography (Invitrogen, USA). The concentration of the purified protein was determined following the Bradford assay procedure (Bradford, M.M., Anal. Biochem., 72:248, 1976).
  • the cell-free extract for crude RbmA from S. lividans TK24/pBS2 was prepared by a similar method and used for the enzyme assay following the dialysis.
  • the concentration of CoCl 2 was maintained at 0.1 mM in the suspension buffer throughout the RbmA manipulations.
  • Standard DOI was prepared using crude BtrC according to the method reported by Kudo and coworkers with slight modifications(Kudo, F. et al., J. Antibiot., 52:559, 1999).
  • reaction product DOI
  • NBHA O-(4-nitrobenzyl) hydroxylamine hydrochloride
  • the assay for RbmA was carried out under identical conditions where the crude BtrC was replaced with the recombinant RbmA from E. coli/ pBSl or from S. lividans TK24/pBS2.
  • the reaction products were derivatized with NBHA and separated by HPLC (SHIMADZU, Japan) at 280 nm.
  • HPLC HPLC
  • a gradient elution was carried out with acetonitrile and acidified water (0.1% trifhioroacetic acid) using C-18 column (MIGHTYSIL -RP-18, Japan) at the flow rate of 1 ml min "1 at 3O 0 C.
  • DOI synthase The activity of DOI synthase was detected by HPLC in a crude enzyme prepared from the 3-day grown transformant; however, no conversion was detected in a crude enzyme from S. lividans TK24/pIBR25 as a control under similar conditions(FIG 5). This indicates that rbmA encodes DOI synthase and it may be involved in the biosynthesis of DOS subunit of Rbm in S. ribosidificus.
  • Example 7 rbml encoding aminoglycoside 3-N-acetyI transferase
  • Rbml was studied in vivo in E. coli (XLl -Blue).
  • the E. coli (XLl-Blue)/pBS3 was resistant to a high concentration of Rbm (800 ⁇ g ml "1 ), Km (300 ⁇ g ml “1 ), Nem (150 ⁇ g ml “1 ), and Apm (200 ⁇ g ml "1 ), whereas the host harboring pET-32a(+) was sensitive to comparatively low concentration (50 ⁇ g ml " ') of these antibiotics.
  • the former bacteria were sensitive to Gm as well as Spn even at a low concentration (30 ⁇ g ml " *).
  • the present invention has an effect to provide DNA sequences of gene cluster participating in Rbm biosynthesis. Futhermore, the present invention is effective to provide DOI synthase as Rbm biosynthetic enzyme and genes encoding the enzyme and aminoglycoside-3-N-acetyltransferase as Rbm biosynthetic enzyme and genes encoding the enzyme. According to the present invention, gene cluster participating in ribostamycin biosynthesis, DOI synthase and AAC(3) can be applied to ribostamycin and similar novel aminoglycosidic antibiotic synthesis.

Landscapes

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

Abstract

La présente invention concerne les enzymes participant à la biosynthèse de la ribostamycine (Rbm) et les gènes correspondants, et plus particulièrement les séquences d'ADN de batterie de gène participant à la biosynthèse de la ribostamycine (Rbm), de la I-dcoxy-scyllo-inosone (DOI) synthase, de l'AAC(3)(aminoglycoside-3-N-acétyltransférase) qui sont des enzymes biosynthétiques de la ribostamycine et des gènes codant pour lesdites enzymes. Selon la présente invention, la batterie de gène participant à la biosynthèse de la ribostamycine (Rbm), la DOI synthase et de l'AAC(3) peut être appliquée efficacement à la préparation de ribostamycine et à une nouvelle synthèse d'aminosides
PCT/KR2005/001801 2004-08-05 2005-06-14 Enzyme impliquee dans la biosynthese de la ribostamycine et les genes correspondants WO2006014047A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2004-0061758 2004-08-05
KR1020040061758A KR100564163B1 (ko) 2004-08-05 2004-08-05 리보스타마이신 생합성에 관여하는 효소 및 그 유전자

Publications (1)

Publication Number Publication Date
WO2006014047A1 true WO2006014047A1 (fr) 2006-02-09

Family

ID=35787316

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2005/001801 WO2006014047A1 (fr) 2004-08-05 2005-06-14 Enzyme impliquee dans la biosynthese de la ribostamycine et les genes correspondants

Country Status (2)

Country Link
KR (1) KR100564163B1 (fr)
WO (1) WO2006014047A1 (fr)

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DATABASE GENPEPT [online] 11 June 2005 (2005-06-11), SUBBA B. ET AL: "Ribostamycin biosynthetic gene cluster in Streptomyces ribosidificus ATCC 21294: Comparison with butirosin biosynthesis", accession no. NCBI Database accession no. (CAG34706) *
DATABASE GENPEPT [online] 11 June 2005 (2005-06-11), SUBBA B. ET AL: "Ribostamycin biosynthetic gene cluster in Streptomyces ribosidificus ATCC 21294: Comparison with butirosin biosynthesis", accession no. NCBI Database accession no. (CAG34718) *
HOSHIKO S. ET AL: "Nucleotide sequence of the ribostamycin phosphotransferase gene and of its control region in Streptomyces ribosidificus", GENE, vol. 68, no. 2, 1988, pages 285 - 296 *
KUDO F. ET AL: "Molecular cloning of the gene for the key carbocycle-forming enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics and its comparison with dehydroquinate synthase", J. ANTIBIOT., vol. 52, no. 6, 1999, pages 559 - 571, XP002990832 *
OWSTON M.A. & SERPERSU E.H.: "Cloning, overexpression, and purification of aminoglycoside antibiotic 3-acetyltransferase-IIIb: conformational studies with bound substrates", BIOCHEMISTRY, vol. 41, no. 35, 2002, pages 10764 - 10770 *

Also Published As

Publication number Publication date
KR20060013001A (ko) 2006-02-09
KR100564163B1 (ko) 2006-03-24

Similar Documents

Publication Publication Date Title
Peschke et al. Molecular characterization of the lincomycin‐production gene cluster of Streptomyces lincolnensis 78‐11
Kharel et al. A gene cluster for biosynthesis of kanamycin from Streptomyces kanamyceticus: comparison with gentamicin biosynthetic gene cluster
Steffensky et al. Identification of the novobiocin biosynthetic gene cluster of Streptomyces spheroides NCIB 11891
May et al. The dhb operon of bacillus subtilisEncodes the biosynthetic template for the catecholic siderophore 2, 3-dihydroxybenzoate-glycine-threonine trimeric ester bacillibactin
Ullrich et al. The biosynthetic gene cluster for coronamic acid, an ethylcyclopropyl amino acid, contains genes homologous to amino acid-activating enzymes and thioesterases
Stohl et al. Zwittermicin A biosynthetic cluster
Garg et al. Molecular characterization and analysis of the biosynthetic gene cluster for the azoxy antibiotic valanimycin
Morbidoni et al. Bacillus subtilis acyl carrier protein is encoded in a cluster of lipid biosynthesis genes
Musiol et al. The AT2 domain of KirCI loads malonyl extender units to the ACPs of the kirromycin PKS
Revill et al. Cloning, characterization, and high-level expression in Escherichia coli of the Saccharopolyspora erythraea gene encoding an acyl carrier protein potentially involved in fatty acid biosynthesis
Fernández-Moreno et al. Streptothricin biosynthesis is catalyzed by enzymes related to nonribosomal peptide bond formation
Petříček et al. Occurrence of two 5-aminolevulinate biosynthetic pathways in Streptomyces nodosus subsp. asukaensis is linked with the production of asukamycin
Priefert et al. Identification and molecular characterization of the acetyl coenzyme A synthetase gene (acoE) of Alcaligenes eutrophus
Subba et al. The ribostamycin biosynthetic gene cluster in Streptomyces ribosidificus: comparison with butirosin biosynthesis
Fu et al. Biosynthesis of 3-hydroxy-5-methyl-O-methyltyrosine in the saframycin/safracin biosynthetic pathway
US7858335B2 (en) Microorganisms and assays for the identification of antibiotics
Wilson et al. Molecular analysis of tlrB, an antibiotic-resistance gene from tylosin-producing Streptomyces fradiae, and discovery of a novel resistance mechanism
WO2006014047A1 (fr) Enzyme impliquee dans la biosynthese de la ribostamycine et les genes correspondants
Singh et al. Precursor for biosynthesis of sugar moiety of doxorubicin depends on rhamnose biosynthetic pathway in Streptomyces peucetius ATCC 27952
Dangel et al. novE and novG act as positive regulators of novobiocin biosynthesis
JP2020500561A (ja) Ｓｔｒｅｐｔｏｍｙｃｅｓ ｆｕｎｇｉｃｉｄｉｃｕｓの遺伝子組換え株におけるエンデュラシジンの産生を増強するための組成物及び方法
QING et al. Genetic organization of a 50-kb gene cluster isolated from Streptomyces kanamyceticus for kanamycin biosynthesis and characterization of kanamycin acetyltransferase
US20050064563A1 (en) Nucleic acids for aminocoumarin biosynthesis
Simkhada et al. Characterization of CalS9 in the biosynthesis of UDP-xylose and the production of xylosyl-attached hybrid compound
KR100488965B1 (ko) 아미노글라이코사이드 인산전이효소를 암호화하는 유전자염기서열 및 아미노산 서열과 그의 항생제 다량 생산에의이용

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NG NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase