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Biosynthesis and molecular genetics of cephamycins

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Abstract

Cephamycin C is produced in a nine steps pathway by the actinomycetes S. clavuligerus and N. lactamdurans. The genes encoding the biosynthesis enzymes are clustered in both microorganisms as well as in the cephabacin producer Lysobacter lactamgenus, a Gram negative bacterium. The clusters of genes include genes encoding enzymes common to the biosynthesis of penicillin and cephalosporin C by the eukaryotic producers Penicillium chrysogenum and Cephalosporiun acremonium and genes for steps specific for the formation of the precursor α-aminoadipic acid as well as for the enzymes involved in the late modification of the cephalosporin intermediates of the pathway. Present are also genes for proteins involved in the export and/or resistance to cephamycin C. In S. clavuligerus a gene encoding a regulatory protein controlling the formation of cephamycin C and clavulanic acid is also present in the cluster.

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References

  • Aharonowitz Y & Demain AL (1979) Nitrogen nutrition and regulation of cephalosporin production in Streptomyces clavuligerus. Can. J. Microbiol. 25: 61-67

    Google Scholar 

  • Aharonowitz Y, Cohen G & Martín JF (1992) Penicillin and cephalosporin biosynthetic genes: structure, organization, regulation, and evolution. Annu. Rev. Microbiol. 46: 461-495

    Google Scholar 

  • Alexander DC & Jensen S (1998) Investigation of the Streptomyces clavuligerus cephamycin C gene cluster and its regulation by the CcaR protein. J. Bacteriol. 180: 4068-4079

    Google Scholar 

  • Arunachalam U, Massey V & Vaidyanathan CS (1992) p-Hydroxyphenylacetate-3-hydroxylase. A two-protein component enzyme. J. Biol. Clem. 267: 25848-25855

    Google Scholar 

  • Baker BJ, Dotzlaf YE & Yeh WK (1991) Deacetoxycephalosporin C hydroxylase of Streptomyces clavuligerus. Purification, characterization, bifunctionality, and evolutionary implication. J. Biol. Chem. 266: 5087-5093

    Google Scholar 

  • Baldwin YE, Adlington RM, Crouch NP & Schofield CJ (1988) The enzymatic conversion of exomethylene cephalosporin C into deacetylcephalosporin C and the role of molecular oxygen in cephalosporin C biosynthesis. Tetrahedron 44: 643-650

    Google Scholar 

  • Borovok I, Landman O, Kreisberg-Zakarin R, Aharonowitz Y & Cohen G (1996) Ferrous active site of isopenicillin N synthase: genetic and sequence analysis of the endogenous ligans. Biochemistry 35: 1981-1987

    Google Scholar 

  • Braña AF, Wolfe S & Demain AL (1985) Ammonium repression of cephalosporin production by Streptomyces clavuligerus. Can. J. Microbiol. 31: 736-743

    Google Scholar 

  • Brewer SJ, Taylor, PM & Turner MIK (1980) An adenosine triphosphate-dependent carbamoylphosphate-3-hydroxymethylcephem O-carbamoyl transferase from Streptomyces clavuligerus. Biochem. J. 185: 555-564

    Google Scholar 

  • Castro JM, Liras P, Cortés J & Martín JF (1985) Regulation of α-aminoadipyl-cysteinyl-valine synthetase, isopenicillin N synthase, isop:nicillin N isomerase and deacetoxycephalosporin C synthase by nitrogen sources in Streptomyces lactamdurans. Appl. Microbiol. Biotechnol. 22: 32-40

    Google Scholar 

  • Castro JM, Liras P, Cortés J & Martín JF (1988) Purification and characterization of the isopenicillin N synthase of Streptomyces lactamdurans. J. Gen. Microbiol. 134: 133-141

    Google Scholar 

  • Chary VK, Fuente JL, Leitão AL, Liras P & Martín JF (1996) Effect of amplification or targeted disruption of the β-lactamase gene of Nocardia lactamdurans on cephamycin biosynthesis. Appl. Microbiol. Biotechnol. 45: 621-628

    Google Scholar 

  • Chary VK, Fuente JL, Liras P & Martín JF (1997) amy as a reporter gene for promotor activity in Nocardia lactamdurans: Comparison of promoters of the cephamycin cluster. Appl. Env. Microbiol. 63: 2977-2982

    Google Scholar 

  • Coque JJR, Liras P, Láiz L & Martín JF (1991a) A gene encoding lysine 6-aminotransferase which forms α-aminoadipic acid, a precursor of β-lactam antibiotics, is located in the cluster of cephamycin biosynthetic genes in Nocardia lactamdurans. J. Bacteriol. 173: 625-6264

    Google Scholar 

  • Coque JJR, Martín JF, Calzada JG & Liras P (1991b) The cephamycin biosynthetic genes pcbAB, encoding a large multidomain peptide synthetase, and pcbC of Nocardia lactamdurans are clustered together in an organization different from the same genes in Acremonium chrysogenum and Penicillium chrysogenum. Mol. Microbiol. 5: 1125-1133

    Google Scholar 

  • Coque JJR, Liras P & Martín JF (1993a) Characterization and expression in Streptomyces lividans of cefD and cefE gene from Nocardia lactamdurans: the organization of the cephamycin gene cluster differs from that in Streptomyces clavuligerus. Mol. Gen. Genet. 236: 453-458

    Google Scholar 

  • Coque JJR, Liras P & Martín JF (1993b) Genes for a β-lactamase, a penicillin-binding protein and a transmembrane protein are clustered with the cephamycin biosynthetic genes in Nocardia lactamdurans. EMBO J. 12: 631-639

    Google Scholar 

  • Coque JJR, Enguita FJ, Martín JF & Liras P (1995a) A two protein component 7α-cephem-methoxylase encoded by two genes of the cephamycin C cluster converts cephalosporin to 7-methoxycephalosporin C. J. Bacteriol. 177: 223-235

    Google Scholar 

  • Coque JJR, Pérez-Llarena FJ, Enguita FJ, de la Fuente IL, Martín JF & Liras P (1995b) Characterization of the cmcH genes of Nocardia lactamdurans and Streptomyces clavuligerus encoding a functional 3′-hydroxymethy cephem O-carbamoyl-transferase from cephamycin biosynthesis. Gene 162: 21-27

    Google Scholar 

  • Coque JR, Enguita FL, Cardoza RE, Martín IF & Liras P (1996a) Characterization of the cefF gene of Nocardia lactamdurans encoding a 3′-methylcephem hydroxylase different from the 7.-cephem hydroxylase. Appl. Microbiol. Biotechnol. 44: 605-609

    Google Scholar 

  • Coque YR, Fuente IL, Liras P & Martín JF (1996b) Overexpression of the Nocardia lactamdurans α-aminoadipyl-cysteinyl-valine synthetase in Streptomyces lividans. Eur. J. Biochem. 242: 264-270

    Google Scholar 

  • Cortés J, Martín JF, Castro JM Laíz L & Liras P (1987) Purification and characterization of a 2-oxoglutarate-linked, ATP-independent deacetoxy-cephalosporin C synthase of Streptomyces lactamdurans. J. Gen. Microbiol. 133: 3165-3174

    Google Scholar 

  • Cortés J, Liras P, Castro JM & Martín JF (1986) Glucose regulation of cephamycin biosynthesis in Streptomyces lactamdurans is exerted on the formation of α-aminoadipyl-cysteinyl-valine and the deacetoxycephalosporin C synthase. J. Gen Microbiol. 132: 1805-1814

    Google Scholar 

  • Deák B, Szabó I, Kalmaczhelyi A., Gal Z, Barabás & Penyge A (1998) Membrane-bound and extracellular β-lactamase production with developmental regulation in Streptomyces griseus NRRL B-2682. Microbiology 144: 2169-2177

    Google Scholar 

  • Doran JL, Leskiw BK, Aippersbach S & Jensen SE (1990a) Isolation and characterization of a β-lactamase-inhibitory protein from Streptomyces clavuligerus and cloning and analysis of the corresponding gene. J. Bacteriol. 172: 4909-4918

    Google Scholar 

  • Doran JL, Leskiw BK, Petrich AK, Westlake DWS & Jensen SE (1990b) Production of Streptomyces clavuligerus isopenicillin N synthase in Escherichia coli using two-cistron expression systems. J. Indust. Microbiol. 5: 197-206

    Google Scholar 

  • Dotzlaf JE & Yeh WK (1987) Copurification and characterization of deacetoxycephalosporin C synthetase/hydroxylase from Cephalosporium acremonium. J. Bacteriol. 169: 1611-1618

    Google Scholar 

  • Enguita FJ, Fuente JL, Martín JF & Liras P (1996a) An inducible expression system of histidine-tagged proteins in Streptomyces lividans for one-step purification by Ni2+ affinity chromatography. FEMS Microbiol. Lett. 137: 135-140

    Google Scholar 

  • Enguita FJ, Liras P, Leitão AL & Martín JF (1996b) Interaction of the two proteins of the methoxylation system involved in cephamycin C biosynthesis. J. Biol. Chem. 271: 33225-33230

    Google Scholar 

  • Enguita FJ, Coque JJR, Liras P & Martín JF (1998) The nine genes of the Nocardia lactamdurans cephamycin cluster are transcribed into large mRNAs from three promoters, two of them located in a bidirectional promoter region. J. Bacteriol. 180: 5409-5494

    Google Scholar 

  • Fernández E, Lombó F, Méndez C & Salas JA (1996) An ABC transporter is essential for resistance to the antitumor agent mithramycin in the producer Streptomyces argillaceus. Mol. Gen. Genet. 251: 692-698

    Google Scholar 

  • Fernández-Moreno MA, Caballero JL, Hopwood DA & Malpartida F (1991) The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the bidA transfer RNA gene of Streptomyces coelicolor. Cell 66: 769-780

    Google Scholar 

  • Fuente JL, Rumbero A, Martín JF & Liras P (1997) Δ-1-Piperideine-6-carboxylate dehydrogenase, a new enzyme that forms α-aminoadipate in Streptomyces clavuligerus and other cephamycin C producing actinomycetes. Biochem. J. 327: 59-64

    Google Scholar 

  • García-Domínguez M, Liras P & Martín JF (1991) Cloning and characterization of the isopenicillin N synthase gene of Streptomyces griseus NRRL3851 and studies of expression and complementation of the cephamycin pathway in Streptomyces clavuligerus. Antimicrob. Agents Chemother. 35: 44-52

    Google Scholar 

  • Horinouchi S, Kito M, Nishiyama M, Furuya K, Hong S-K, Miyake K & Beppu T (1990) Primary structure of AfsR, a global regulatory protein for secondary metabolite formation in Streytomyces coelicolor A3(2). Gene 95: 49-56

    Google Scholar 

  • Ingram C, Brawner M, Youngman P & Westpheling J (1989) xylE functions as an efficient reporter gene in Streptomyces spp.: use for the study of galP1, a catabolite-controlled promoter. J. Bacteriol. 171: 6617-6624

    Google Scholar 

  • Jensen SE, Leskiw BK, Vining LC, Aharonowitz Y & Westlake DWS (1986) Purification of isopenicillin N synthase from Streptomyces clavuligerus. Can. J. Microbiol. 32: 953-958

    Google Scholar 

  • Jensen SE, Westlake DWS & Wolfe S (1985) Deacetoxycephalosporin C synthetase and deacetoxycephalosporin C hydroxylase are two separate enzymes in Streptomyces clavuligerus. J. Antibiot. 38: 263-265

    Google Scholar 

  • Jensen SE, Wong A, Rollins MJ & Westlake DWS (1990) Purification and partial characterization of δ(L-α-aminoadipyl)-L-cysteinyl-valine synthetase from Streptomyces clavuligerus. J. Bacteriol. 172: 7269-7271

    Google Scholar 

  • Kern BA & Inamine E (1981) Cystathionine-γ-lyase activity in the cephamycin C producer Streptomyces lactamdurans. J. Antibiot. 34: 583-589

    Google Scholar 

  • Kern BA, Hendlin D & Inamine EE (1980) L-lysine ε-aminotransferase involved in cephamycin C synthesis in Streptomyces lactamdurans. Antimicrob. Agents Chemother. 17: 676-685

    Google Scholar 

  • Kimura H, Miyashita H & Sumino Y (1996) Organization and expression in Pseudomonas putida of the gene cluster involved in cephalosporin biosynthesis from Lysobacter lactamgenus YK90. Appl. Microbiol. Biotechnol. 45: 490-501

    Google Scholar 

  • Kleinkauf H & von Döhren H (1996) A nonribosomal system of peptide biosynthesis. Eur. J. Biochem. 236: 335-351

    Google Scholar 

  • Kovacevic S & Miller JR (1991) Cloning and sequencing of the β-lactam hydroxylase gene (cefF) from Streptomyces clavuligerus: gene duplication may have led to separate hydroxylase and expandase activities in the actinomycetes. J. Bacteriol. 173: 398-400

    Google Scholar 

  • Kovacevic S, Weigel BJ, Tobin MB, Ingolia TD & Miller YR (1989) Cloning, characterization, and expression in Escherichia coli of the Streptomyces clavuligerus gene encoding deacetoxycephalosporin C synthetase. J. Bacteriol. 171: 754-760

    Google Scholar 

  • Kovacevic S, Tobin MB & Miller JR (1990) The β-lactam biosynthesis genes for isopenicillin N epimerase and deacetoxycephalosporin C synthetase are expressed from a single transcript in Streptomyces clavuligerus. J. Bacteriol. 172: 3952-3958

    Google Scholar 

  • Láiz L, Liras P, Castro JM & Martín JF (1990) Purification and characterization of the isopenicillin N epimerase from Nocardia lactamdurans. J. Gen. Microbiol. 136: 663-671

    Google Scholar 

  • Landan G, Cohen G, Aharonowitz Y, Shuali Y, Graur D & Shiffman D (1990) Evolution of isopenicillin N synthase genes may have involved horizontal gene transfer. Mol. Biol. Evol. 7: 399-406

    Google Scholar 

  • Landman O, Borovok I, Aharonowitz Y & Cohen G (1977) The glutamine ligand in the ferrous iron active site of isopenicillin N synthase of Streptomyces jumonjinensis is not essential for catalysis. FEES Lett. 405: 172-174

    Google Scholar 

  • Leskiw BK, Lawlor EJ, Fernández-Abalos JM & Chater KF (1991) TTA codons in some genes prevent their expression in a class of developmental, antibiotic negative, Streptomyces mutants. Proc. Natl. Acad. Sci. USA 88: 2461-2465

    Google Scholar 

  • Madduri K, Stuttard C & Vining LC (1991) Cloning and location of a gene governing lysine ε-aminotransferase, an enzyme initiating β-lactam biosynthesis in Streptomyces sp. J. Bacteriol. 173: 985-988

    Google Scholar 

  • Martín JF (1998) New aspects of gene and enzymes for β-lactam antibiotic biosynthesis. Appl. Microbiol. Biotechnol. 50: 1-15

    Google Scholar 

  • Martín JF & Gutiérrez S (1995) Genes for β-lactam antibiotic biosynthesis. Antonie van Leeuwenhoek 67: 181-200

    Google Scholar 

  • Martín JF & Liras P (1989) Enzymes involved in penicillin, cephalosporin and cephamycin biosynthesis. In: Fiechter A (Ed) Advances in Biochemical Engineering/Biotechnology, Vol. 39 (pp 153-187). Springer Verlag, Berlin

    Google Scholar 

  • Martín JF, Gutiérrez S, Fernández FJ, Velasco J, Fierro F, Marcos AT & Kosalkova K (1994) Expression of genes and processing of enzymes for the biosynthesis of penicillins and cephalosporins. Antonie van Leeuwenhoek 65: 227-243

    Google Scholar 

  • Nagasawa T, Kanzaki H & Yamada H (1984) Cystathionine-γ-lyase of Streptomyces phaeochromogenes. J. Biol. Chem. 259: 10393-10403

    Google Scholar 

  • Narva KE & Feitelson JS (1990) Nucleotide sequence and transcriptional analysis of the redD locus of Streptomyces coelicolor A(3)2. J. Bacteriol. 172: 326-333

    Google Scholar 

  • Ogawara H & Horikawa S (1980) Penicillin-binding proteins of Streptomyces cacaoi, Streptomyces olivaceus and Streptomyces clavuligerus. Antimicrob. Agents Chemother. 17: 1-7

    Google Scholar 

  • Ono H, Nozaki Y, Katayama N & Okazaki H (1984) Cephabacins, new cephem antibiotics of bacterial origin. I. Discovery and taxonomy of the producing organisms and fermentation. J. Antibiot. 37:1528-1534

    Google Scholar 

  • Paradkar AS, Aidoo KA, Wong A & Jensen SE (1996) Molecular analysis of a β-lactam resistance gene encoded within the cephamycin gene cluster of Streptomyces clavuligerus. J. Bacteriol. 178: 6266-6274

    Google Scholar 

  • Paradkar AS, Jensen SE & Mosher RH (1997) Comparative genetics and molecular biology of β-lactam biosynthesis. In: Strohl WR (Ed) Biotechnology of Antibiotics. Marcel Dekker, New York

    Google Scholar 

  • Paradkar AS, Aidoo KA & Jensen SE (1998) A pathway-specific transcriptional activator regulates late steps of clavulanic acid biosynthesis in Streptomyces clavuligerus. Mol. Microbiol. 27: 831-843

    Google Scholar 

  • Paulsen IT, Brown MR & Skurray RA (1996) Proton-dependent multidrug eflux systems. Microbiol. Rev. 60: 575-608

    Google Scholar 

  • Pérez-Llarena FJ (1997) Caracterización de la agrupación de genes de cefamicina C en Streptomyces clavuligerus. Ph.D. Thesis, University of León, Spain

    Google Scholar 

  • Pérez-Llarena FJ, Liras P, Rodríguez-García A & Martín IF (1997) A regulatory gene (ccaR) required for cephamycin and clavulanic acid production in Streptomyces clavuligerus: Amplification results in overproduction of both β-lactam compounds. J. Bacteriol. 179: 2053-2059

    Google Scholar 

  • Pérez-Llarena FJ, Rodríguez-García A, Enguita FJ, Martín JF, Liras P (1998) The pcd gene encoding piperideine-6-carboxylate de-hydrogenase involved in biosynthesis of α-aminoadipic acid is located in the cephamycin cluster of S. clavuligerus. J. Bacteriol. 180: 4753-4756.

    Google Scholar 

  • Pérez-Llarena FJ, Martín JF, Coque JJR, Fuente JL, Galleni M, Frère J-M & Liras P (1996) The bla gene of the cephamycin cluster of Streptomyces clavuligerus encodes a class A β-lactamase of low enzymatic activity. J. Bacteriol. 179: 6035-6040

    Google Scholar 

  • Pérez-Redondo R, Rodríguez-García A, Martín JF & Liras P (1998) The claR gene of Streptomyces clavuligerus, encoding a LysR-type regulatory protein controlling clavulanic acid biosynthesis, is linked to the clavulanate-9-aldehyde reductase (car) gene. Gene 211: 311-321

    Google Scholar 

  • Petrich AK, Wu X, Roy KL & Jensen SE (1992) Transcriptional analysis of the isopenicillin N synthase encoding gene of Streptomyces clavuligerus. Gene 111: 77-84

    Google Scholar 

  • Petrich AK, Leskiw BK, Paradkar AS & Jensen SE (1994) Transcriptional mapping of the genes encoding the early enzymes of the cephamycin biosynthetic pathway of Streptomyces clavuligerus. Gene 142:41-48

    Google Scholar 

  • Roach PL, Clifton IJ, Fülöp V, Harlos K, Barton GJ, Hajdu J, Andersson I, Schofield CJ & Baldwin JE (1995) Crystal structure of isopenicillin N synthase is the first from a new structural family of enzymes. Nature 375: 700-704

    Google Scholar 

  • Romero J, Liras P & Martín JF (1988) Isolation and biochemical characterization of Streptomyces clavuligerus mutants in the biosynthesis of clavulanic acid and cephamycin C. Appl. Microbiol. Biotechnol. 27: 510-516

    Google Scholar 

  • Samson SM, Dotzlaf YE, Slisz ML, Becker GW, Van Frank RM, Veal LE, Yeh WK, Miller YR. Queener SW & Ingolia TD (1987) Cloning and expression of the fungal expandase/hydroxylase gene involved in cephalosporin biosynthesis. Bio/Technology 5: 1207-1214

    Google Scholar 

  • Schwecke T, Aharonowitz Y, Palissa H, von Döhren H, Kleinkauf H & von Liempt H (1992) Enzymatic characterization of the multifunctional enzyme δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase from Streptomyces clavuligerus. Eur. J. Biochem 205: 687-694

    Google Scholar 

  • Strynadka NCJ, Jensen S, Johns K, Blanchard H, Page M, Matagne A, Frére JM & James MNG (1994) Structural and kinetic characterization of a β-lactamase-inhibitor protein. Nature 368: 657-660

    Google Scholar 

  • Stutzman-Engwall KJ, Often, SL & Hutchinson CR (1992) Regulation of secondary metabolism in Streptomyces spp. and overproduction of daunorubicin in Streptomyces peucetius. J. Bacteriol. 174: 144-154

    Google Scholar 

  • Tobin MB, Kovacevik S, Madduri K, Hoskins JA, Skatrud PL, Vining LC, Stuttards C & Miller YR (1991) Localization of the lysine ε-aminotransferase (lat) and δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine synthetase (pcbAB) genes from Streptomyces clavuligerus and production of lysine α-aminotransferase activity in Escherichia coli. J. Bacteriol. 173: 6223-6229

    Google Scholar 

  • Usui S & Yu CA (1989) Purification and properties of isopenicillin N epimerase from Streptomyces clavuligerus. Bioch. Biophys. Acta 999: 78-85

    Google Scholar 

  • Van Liempt H, von Döhren H & Kleinkauf H (1989) δ-(L-α-aminoadipyl)-L-cysteinyl-valine synthetase from Aspergillus nidulans. J. Biol. Chem. 264: 3680-3684

    Google Scholar 

  • Wietzorrek A & Bibb M (1997) A novel family of proteins that regulate antibiotic production in streptomycetes appears to contain an OmpR-like DNA-binding fold. Mol. Microbiol. 25: 1181-1184

    Google Scholar 

  • Xiao X, Bowers RJ, Shin H-S, Wolfe S & Demain AL (1991a) Non-radioactive assay and HPLC analysis of cephalosporin C 7-α-methoxylation by cell-free extracts of Streptomyces clavuligerus. Appl. Microbiol. Biotechnol. 35: 793-797

    Google Scholar 

  • Xiao X, Wolfe S & Demain AL (1991b) Purification and characterization of cephalosporin 7α-hydroxylase from Streptomyces clavuligerus. Biochem. J. 280: 471-474

    Google Scholar 

  • Yu H, Serpe E, Romero J, Coque JJ, Maeda K, Oelgeschláger M, Hintermann G, Liras P, Martín JF, Demain AL & Piret J (1994) Possible involvement of the lysine ε-aminotransferase gene (lat) in the expression of the genes encoding ACV synthetase (pcbAB) and isopenicillin N synthase (pcbC) in Streptomyces clavuligerus. Microbiology 140: 3367-3377

    Google Scholar 

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  1. Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain

    Paloma Liras

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Liras, P. Biosynthesis and molecular genetics of cephamycins. Antonie Van Leeuwenhoek 75, 109–124 (1999). https://doi.org/10.1023/A:1001804925843

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