Studies on the biosynthesis of 16-membered macrolide antibiotics using carbon-13 nuclear magnetic resonance spectroscopy

Ōmura, Satoshi; Tanaka, Hideo; Nakagawa, Akira; Miyazawa, J.; Piriou, F.; Lukacs, Gabor

doi:10.1021/bi00632a009

Cited by 51 publications

(30 citation statements)

References 17 publications

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“…1) as well as at C-11/12, C-13/14, and C-15/16 in the manner described for [3-13C]ethylmalonate. Curiously, though, there did not appear to be enrichment at C-3/4 and C-7/8, as would be expected (6,8,9).…”

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confidence: 69%

“…The complex incorporation patterns reported for C4 units (6,8,9) dictate caution when interpreting the group III labeling data presented here. Nevertheless, the fact that the unlabeled C2 (acetate) and C3 (isoleucine) subunit precursors were present at concentrations 32 times greater than the group III test compounds suggests that leucine and phenylalanine indeed labeled tylactone as the C4 (butyrate) subunit.…”

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confidence: 84%

“…Propionate was incorporated into CO2 and cells in a fashion similar to a-ketoglutarate and succinate (Fig. 6 (5,(7)(8)(9). Since the interconversion of propionyl-CoA and methylmalonyl-CoA has been demonstrated in S. fradiae (10), one can postulate common sources for these two tylactone precursors.…”

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confidence: 95%

“…Omura et al (6), using 13C enrichment, showed that tylactone was formed from five propionate molecules, two acetate molecules, and one butyrate molecule. Subsequent inhibition studies with cerulenin, an inhibitor of fatty acid synthetase, showed that tylactone and related macrolide aglycone molecules were assembled in a manner similar to fatty acid biosynthesis (5,(7)(8)(9). The data of Omura et al (5)(6)(7)(8)(9) suggested that the precursors for tylactone biosynthesis were incorporated in the following manner: propionate as one propionyl-coenzyme A (CoA) (primer) and four methylmalonyl-CoA molecules, acetate as two malonyl-CoA molecules, and butyrate as one ethylmalonylCoA molecule.…”

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confidence: 99%

“…Subsequent work led Omura and his co-workers (8,9) to propose separate pathways for the conversion of butyrate and ethylmalonate to methylmalonyl-CoA for incorporation into the C3 subunit portions of tylactone. While exploring the possible pathways leading to methylmalonyl-CoA, Omura et al (8,9) utilized [1,3,1'-13C]ethylmalonate and found enrichment at C-5 and C-19 ( Fig. 1) as well as at C-11/12, C-13/14, and C-15/16 in the manner described for [3-13C]ethylmalonate.…”

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confidence: 99%

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Incorporation of amino acid-derived carbon into tylactone by Streptomyces fradiae GS14

Dotzlaf¹,

Metzger²,

Foglesong³

1984

Antimicrob Agents Chemother

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Washed cells from 72-h cultures of Streptomyces fradiae GS14 were used to examine the distribution of radiolabel from 14C-amino acids and related compounds into tylactone, C02, and cells. Test compounds were categorized according to products of their oxidative degradation. Those compounds known to produce propionyl-coenzyme A by direct catabolic oxidation were designated as group I. Group II included those compounds oxidized to methylmalonyl-coenzyme A via succinyl-coenzyme A and the tricarboxylic acid cycle. Group III contained compounds known to be oxidized to acetoacetyl-coenzyme A. The total amount of label recovered after 60 min ranged from 3 to 65%. Although label from all test compounds except proline (group II) and lysine (group III) was incorporated into tylactone after 60 min, label from group I and group III compounds was incorporated at levels five times greater than label from group II compounds. From 55 to 75% of the recovered label from propionate (I), asparagine (II), glutamine (II), glutamate (II), aketoglutarate (II), and succinate (II) was recovered as 14 C02. From 75 to 95% of the recovered label from the remaining compounds tested was located in the cells. Based on the data, a pathway for the role of amino acids in the biosynthesis of tylactone is proposed.Tylactone (Fig. 1) is the initial lactone precursor to tylosin, an antibiotic produced commercially by Streptomyces fradiae (1-3). Omura et al. (6), using 13C enrichment, showed that tylactone was formed from five propionate molecules, two acetate molecules, and one butyrate molecule. Subsequent inhibition studies with cerulenin, an inhibitor of fatty acid synthetase, showed that tylactone and related macrolide aglycone molecules were assembled in a manner similar to fatty acid biosynthesis (5, 7-9). The data of suggested that the precursors for tylactone biosynthesis were incorporated in the following manner: propionate as one propionyl-coenzyme A (CoA) (primer) and four methylmalonyl-CoA molecules, acetate as two malonyl-CoA molecules, and butyrate as one ethylmalonylCoA molecule.In this study, the role of amino acids as sources for the tylactone precursors was examined by using a tylA mutant of S. fradiae (1). This strain was blocked in the production of the tylosin sugars and therefore produced tylactone as its product (1, were grown in a complex medium as described previously (1).Preparation of washed cell suspensions. After 72 h of incubation, the cells from 300 ml of broth were removed by centrifugation at 16,000 x g for 5 min in a Sorvall RC-5 centrifuge. Cell pellets were washed twice with 300 ml of 0.1 M potassium phosphate buffer, pH 7.0. Approximately 6.5 g of wet cells were suspended in the buffer to a volume of 50 ml, and portions were diluted 10-fold with buffer to yield final cell suspensions containing 1.2 to 1.7 mg of dry cells per ml.Uptake and distribution reactions. Reactions were carried out in 35-ml flint glass bottles (Kimble, Toledo, Ohio) that were closed with gray butyl rubber stoppers (West Rubber Co., Phoenix...

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confidence: 69%

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confidence: 84%

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confidence: 95%

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confidence: 99%

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confidence: 99%

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Incorporation of amino acid-derived carbon into tylactone by Streptomyces fradiae GS14

Dotzlaf¹,

Metzger²,

Foglesong³

1984

Antimicrob Agents Chemother

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Seven‐, Eight‐Membered and Larger Heterocyclic Rings and Their Fused Derivatives

Diana

Cirrincione

2015

Biosynthesis of Heterocycles

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The Biosynthesis of Aliphatic Polyketides

Staunton

Wilkinson²

1998

Biosynthesis

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Aliphatic polyketides are a large family of natural products which exhibit an impressive range of biological activities. They are synthesised by a common pathway in which units derived from acetate, propionate and butyrate are condensed onto a growing chain, which is initiated from a range of structurally varied carboxylic acid starter units, in a process closely resembling fatty acid biosynthesis. The intermediates remain bound to the polyketide synthase (PKS) throughout multiple rounds of chain extension, and to a variable degree, reduction of the newly formed b-keto functionality. It is the manner in which each different PKS controls the number and type of extender units added, and the extent and stereochemistry of reduction at each cycle which gives rise to the diverse structural variation seen in this family of natural products. Furthermore, the aglycone product of the PKS may be modified by post-PKS enzymes, including glycosidases, methyltransferases, acylases and oxidative enzymes, to produce still greater diversity. In this article the development of the field will be exemplified by a detailed discussion of the archetypal example of erythromycin A biosynthesis. This will then be followed by further discussion of other relevant areas of research in this field.

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Studies on the biosynthesis of 16-membered macrolide antibiotics using carbon-13 nuclear magnetic resonance spectroscopy

Cited by 51 publications

References 17 publications

Incorporation of amino acid-derived carbon into tylactone by Streptomyces fradiae GS14

Incorporation of amino acid-derived carbon into tylactone by Streptomyces fradiae GS14

Seven‐, Eight‐Membered and Larger Heterocyclic Rings and Their Fused Derivatives

The Biosynthesis of Aliphatic Polyketides

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