Abstract
The biosynthesis of clavulanic acid and related clavam metabolites is only now being elucidated. Understanding of this pathway has resulted from a combination of both biochemical studies of purified biosynthetic enzymes, and molecular genetic studies of the genes encoding these enzymes. Clavulanic acid biosynthesis has been most thoroughly investigated in Streptomyces clavuligerus where the biosynthetic gene cluster resides immediately adjacent to the cluster of cephamycin biosynthetic genes. A minimum of eight structural genes have been implicated in clavulanic acid biosynthesis, although more are probably involved. While details of the early and late steps of the pathway remain unclear, synthesis proceeds from arginine and pyruvate, as the most likely primary metabolic precursors, through the monocyclic β-lactam intermediate, proclavaminic acid, to the bicyclic intermediate, clavaminic acid, which is a branch point leading either to clavulanic acid or the other clavams. Conversion of clavaminic acid to clavulanic acid requires side chain modfication as well as inversion of ring stereochemistry. This stereochemical change occurs coincident with acquisition of the β-lactamase inhibitory activity which gives clavulanic acid its therapeutic and commercial importance. In contrast, the other clavam metabolites all arise from clavaminic acid with retention of configuration and lack β-lactamase inhibitory activity.
Similar content being viewed by others
References
Aidoo KA, Wong A, Alexander DC, Rittammer RAR & Jensen SE (1994) Cloning, sequencing and disruption of a gene from Streptomyces clavuligerus involved in clavulanic acid biosynthesis. Gene 147: 41–46
Bachmann BO, Li R & Townsend CA (1998) β-Lactam synthetase: a new biosynthetic enzyme. Proc. Natl. Acad. Sci. USA 95: 9082–9086
Baggaley KH, Brown AG & Schofield CJ (1997) Chemistry and biosynthesis of clavulanic acid and other clavams. Natural Products Reports 309–333
Baggaley KH, Elson SW, Nicholson NH & Sime JT (1990) Studies on the biosynthesis of clavulanic acid. Part 4. Synthetic routes to the monocyclic β-lactam precursor, proclavaminic acid. J. Chem. Soc. Perkin Trans. I. 1513–1520
Bailey CR, Butler MJ, Normansell ID, Rowlands RT & Winstanley DJ (1984) Cloning a Streptomyces clavuligerus genetic locus involved in clavulanic acid biosynthesis. Bio/Technology 2: 808–811
Baldwin JE, Adlington RM, Bryans JS, Bringhen AO, Coates JB, Crouch NP, Lloyd MD, Schofield CJ, Elson SW, Baggaley KH, Cassels R & Nicholson N (1991) Isolation of dihydroclavaminic acid, an intermediate in the biosynthesis of clavulanic acid. Tetrahedron 47: 4089–4100
Baldwin JE, Lloyd MD, Wha-Son B, Schofield CJ, Elson SW, Baggaley KH & Nicholson NH (1993) A substrate analogue study on clavaminic acid synthase: possible clues to the biosynthetic origin of proclavaminic acid. J. Chem. Soc., Chem. Commun. 500–502
Box SJ (1977) Clavulanic acid and its salts by fermentation. Chem. Abstr. 87: 4031e
Brown AG, Butterworth D, Cole, Hanscombe G, Hood JD, Reading C & Robinson GN (1976) Naturally occurring beta-lactamase inhibitors with antibacterial activity. J. Antibiot. 29: 668–669
Brown D, Evans JR & Fletton RA (1979) Structures of three novel β-lactams isolated from Streptomyces clavuligerus. J. Chem. Soc., Chem. Commun. 282–283
Egan LA, Busby RW, Iwata-Reuyl D & Townsend CA (1997) Probable role of clavaminic acid as the terminal intermediate in the common pathway to clavulanic acid and the antipodal clavam metabolites. J. Am. Chem. Soc. 119: 2348–2355
Elson SW, Baggaley KH, Davison M, Fulston M, Nicholson NH, Risbridger GD & Tyler JW (1993a) The identification of three new biosynthetic intermediates and one further biosynthetic enzyme in the clavulanic acid pathway. J. Chem. Soc., Chem. Commun. 1212–1214
Elson SW, Baggaley KH, Fulston M, Nicholson NH, Tyler JW, Edwards J, Holms H, Hamilton I & Mousdale DM (1993b) Two novel arginine derivatives from a mutant of Streptomyces clavuligerus. J. Chem. Soc., Chem. Commun. 1211–1212
Elson SW, Baggaley KH, Gillett J, Holland S, Nicholson NH, Sime JT & Woroniecki SR (1987) Isolation of two novel intracellular ß-lactams and a novel dioxygenase cyclising enzyme from Streptomyces clavuligerus. J. Chem. Soc., Chem. Commun. 1736–1738
Hodgson JE, Fosberry AP, Rawlinson NS, Ross HNM, Neal RJ, Arnell JC, Earl AJ & Lawlor EJ (1995) Clavulanic acid biosynthesis in Streptomyces clavuligerus: gene cloning and characterization. Gene 166: 49–55
Janc JW, Egan LA & Townsend CA (1993) Emerging evidence for a shared biosynthetic pathway among clavulanic acid and the structurally diverse clavam metabolites. Bioorg. Med. Chem. Lett. 3: 2313–2316
Janc JW, Egan LA & Townsend CA (1995) Purification and characterization of clavaminate synthase from Streptomyces antibioticus: A multifunctional enzyme of clavam biosynthesis. J. Biol. Chem. 270: 5399–5404
Jensen SE, Alexander DC, Paradkar AS & Aidoo KA (1993) Extending the β-lactam biosynthetic gene cluster in Streptomyces clavuligerus, p. 169–176. In: Baltz RH, Hegeman GD & Skatrud PL (Eds) Industrial Microorganisms: Basic and Applied Molecular Genetics. American Society for Microbiology, Washington, DC
Kawamura Y, Kondo E, Matsumoto K, Tsuji N & Tawara K (1987) Clavam antibiotic CA-31. Chem. Abstr. 106: 212565v
Kawamura Y, Kondo E, Matsumoto K, Tsuji N & Tawara K (1987) Clavam antibiotics CA-146A and 146B. Chem. Abstr. 107: 76085e
Kitano K, Kintaka K & Katamoto K (1979) Clavulanic acid production by Streptomyces katsurahamanus. Chem. Abstr. 90: 119758b
Marsh NE, Chang MD-T & Townsend CA (1992) Two isozymes of clavaminate synthase central to clavulanic acid formation: cloning and sequencing of both genes from Streptomyces clavuligerus. Biochemistry 31: 1268–12657
McGowan SJ, Sebaihia m, Porter LE, Stewart GSAB, Williams P, Bycroft BW & Salmond GPC (1996) Analysis of bacterial carbapenem antibiotic production genes reveals a novel β-lactam biosynthesis pathway. Mol. Microbiol. 22: 415–426
Naegeli HU, Loosli HR & Nussbaumer A (1986) Clavamycins, new clavam antibiotics from two variants of Streptomyces hygroscopicus. II. Isolation and structures of clavamycins A, B and C from Streptomyces hygroscopicus NRRL 15846 and of clavamycin D, E and F from Streptomyces hygroscopicus NRRL 15879. J. Antibiot. 39: 516–524
Nagarajan R, Boeck LD, Gorman M, Hamill RL Higgens CE, Hoehn MM, Stark WM & Whitney JG (1971) β-Lactam antibiotics from Streptomyces. J. Am. Chem. Soc. 93: 2308–2310
Nicholson NH, Baggaley KH, Cassels R, Davison M, Elson SW, Fulston M, Tyler JW & Woroniecki SR (1994) Evidence that the immediate biosynthetic precursor of clavulanic acid is its N-aldehyde analogue. J. Chem. Soc., Chem. Commun. 1281–1282
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
Perez-Llarena F, Liras P, Rodríguez-García A & Martín JF (1997) A regulatory gene (ccaR) required for cephamycin and clavulanic acid production in Streptomyces clavuligerus: amplification results in overproduction of both beta-lactam compounds. J. Bacteriol. 179: 2053–2059
Peter H, Rabenhorst J, Rohl F & Zahner H (1985). In: Ishigami J (Ed) Recent Advances in Chemotherapy. University of Tokyo Press, Tokyo, p. 237–238
Pitlik J & Townsend CA (1997) The fate of [2, 3, 3-2H3, 1,2-13C2]-D, L-glycerate in clavulanic acid biosynthesis. J. Chem. Soc., Chem. Commun. 225–226
Pruess DL & Kellett m (1983) Ro–22–5417, a new clavam antibiotic from Streptomyces clavuligerus. I. Discovery and biological activity. J. Antibiot. 36: 208–212
Romero J, Liras P & Martin JF (1986) Utilization of ornithine and arginine as specific precursors of clavulanic acid. Appl. Environ. Microbiol. 52: 892–897
Salowe SP, Krol WJ, Iwata-Reuyl D & Townsend CA (1991) Elucidation of the order of oxidations and identification of an intermediate in the multistep clavaminate synthase reaction. Biochemistry 30: 2281–2292
Salowe SP, Marsh EN & Townsend CA (1990) Purification and characterization of clavaminate synthase from Streptomyces clavuligerus: an unusual oxidative enzyme in natural product biosynthesis. Biochemistry 29: 6499–6508
Sanraku-Ocean Co. Ltd. (1981) Clavulanic acid. Chem Abstr 94: 137803z
Seki C, Den K, Marunaka T, Miyake Y, Minami Y Kajitani A & Ishida N (1987) Antitumor and antifungus G0069C substance production by Streptomyces species. JP 61, 268, 685. Chem. Abstr. 106: 194774f
Thirkettle JE, Baldwin JE, Edwards J, Griffin JP & Schofield CJ (1997) The origin of the β-lactam carbons of clavulanic acid. J. Chem. Soc., Chem Commun. 1025-1026
Valentine BP, Bailey CR, Doherty A, Morris J, Elson SW, Baggaley KH & Nicholson NH (1993) Evidence that arginine is a later metabolic intermediate than ornithine in the biosynthesis of clavulanic acid by Streptomyces clavuligerus. J. Chem. Soc., Chem. Commun. 1210-1211
Walters NJ, Barton B & Earl AJ (1994) Novel compounds. International patent: WO 94/18326-A1
Wanning M, Zahner H, Krone B & Zeeck A (1981) Ein neues antifungisches β-lactam antibioticum der clavam-reihe. Tetrahedron Lett. 22: 2539-2540
Ward JM & Hodgson JE (1993) The biosynthetic genes for clavulanic acid and cephamycin production occur as a 'super-cluster’ in three Streptomyces. FEMS Microbiol. Lett. 110: 239-242
Wietzorrek A & Bibb M (1997) A novel family of proteins that regulates antibiotic production in streptomycetes appears to contain an OmpR-like DNA-binding fold. Mol. Microbiol. 25: 1181-1184
Zhang W, Qi C, Tian J, Xie m, Chen W & Sun J (1991) New clavam antibiotic G0069. Chem. Abstr. 115: 269958r
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Jensen, S.E., Paradkar, A.S. Biosynthesis and Molecular Genetics of Clavulanic Acid. Antonie Van Leeuwenhoek 75, 125–133 (1999). https://doi.org/10.1023/A:1001755724055
Issue Date:
DOI: https://doi.org/10.1023/A:1001755724055