Two distinctive O-methyltransferases catalyzing penultimate and terminal reactions of macrolide antibiotic (tylosin) biosynthesis. Substrate specificity, enzyme inhibition, and kinetic mechanism.

Kreuzman, A. J.; Turner, J. R.; Yeh, Wu-Kuang

doi:10.1016/s0021-9258(19)37634-3

Cited by 47 publications

(16 citation statements)

References 14 publications

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“…Similarly, 2‴- O -demethyltylosin ( 1 ) was converted to tylosin by YJ028/pYJ766 at a conversion yield of approximately 50% (Figure C). These results prove that TylE is able to catalyze the C2‴- O -methylation of 2‴- O -demethyldesmycosin ( 2 ) and 2‴- O -demethyltylosin ( 1 ) in contrast to the previous studies …”

Section: Resultssupporting

confidence: 65%

“…Therefore, it is likely that the newly found biosynthetic routes to desmycosin and tylosin via 2‴- O -demethyldesmycosin ( 2 ) and 2‴- O -demethyltylosin ( 1 ), respectively, are minor tylosin biosynthetic pathways. This is probably the reason why previous studies , using the TylE and TylF purified from S. fradiae did not detect the production of these minor biosynthetic intermediates.…”

Section: Resultsmentioning

confidence: 89%

“…It has long been known that TylE-catalyzed C2‴- O -methylation of the 6-deoxy- d -allose moiety normally precedes the TylF-catalyzed C3‴- O -methylation. ,, However, to confirm whether TylF can catalyze C3‴- O -methylation of the 6-deoxy- d -allose moiety of demethyllactenocin instead of the d -javose in lactenocin, we constructed S. venezuelae mutant strains which can transfer TDP-6-deoxy- d -allose to the exogenously fed O -mycaminosyltylonolide and further modify either the 6-deoxy- d -allose moiety via 2‴- or 3‴- O -methylation as well as both. S. venezuelae YJ028, in which the entire pikromycin biosynthetic gene cluster was deleted from the wild-type S. venezuelae ATCC 15439, was used for the construction of mutant strains.…”

Section: Resultsmentioning

confidence: 99%

“…The two S -adenosyl- l -methionine (SAM)-dependent O -methyltransferases TylE and TylF were purified from S. fradiae and shown to sequentially methylate the C2‴- and C3‴-hydroxy groups of the 6-deoxy- d -allose moiety of demethylmacrocin and demethyllactenocin, yielding tylosin and desmycosin via macrocin and lactenocin, respectively (Figure ). , All the d -mycinose biosynthetic genes of S. fradiae were also cloned and sequenced . It has been shown using purified enzymes that both TylE and TylF possess high substrate specificity and that the C2‴- O -methylation of the 6-deoxy- d -allose moiety catalyzed by TylE generating d -javose (C2‴- O -methylated 6-deoxy- d -allose) is a prerequisite for the subsequent C3‴- O -methylation by TylF needed to generate d -mycinose (C3‴- O -methylated d -javose). , This methylation order is consistent with the biosynthetic pathway of mycinamicin IV reported recently.…”

mentioning

confidence: 99%

“… , All the d -mycinose biosynthetic genes of S. fradiae were also cloned and sequenced . It has been shown using purified enzymes that both TylE and TylF possess high substrate specificity and that the C2‴- O -methylation of the 6-deoxy- d -allose moiety catalyzed by TylE generating d -javose (C2‴- O -methylated 6-deoxy- d -allose) is a prerequisite for the subsequent C3‴- O -methylation by TylF needed to generate d -mycinose (C3‴- O -methylated d -javose). , This methylation order is consistent with the biosynthetic pathway of mycinamicin IV reported recently. The purified recombinant MycE, a TylE homologue, methylates the C2″-hydroxy group of 6-deoxy- d -allose, converting mycinamicin VI to mycinamicin III, and the TylF homologue MycF catalyzes the second C3″- O -methylation of d -javose giving mycinamicin IV .…”

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confidence: 99%

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Characterization of the Two Methylation Steps Involved in the Biosynthesis of Mycinose in Tylosin

et al. 2016

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The S-adenosyl-l-methionine-dependent O-methyltransferases TylE and TylF catalyze the last two methylation reactions in the tylosin biosynthetic pathway of Streptomyces fradiae. It has long been known that the TylE-catalyzed C2‴-O-methylation of the 6-deoxy-d-allose bound to demethylmacrocin or demethyllactenocin precedes the TylF-catalyzed C3‴-O-methylation of the d-javose (C2‴-O-methylated 6-deoxy-d-allose) attached to macrocin or lactenocin. This study reveals the unexpected substrate promiscuity of TylE and TylF responsible for the biosynthesis of d-mycinose (C3‴-O-methylated d-javose) in tylosin through the identification of a new minor intermediate 2‴-O-demethyldesmycosin (2; 3‴-methyl-demethyllactenocin), which lacks a 2‴-O-methyl group on the mycinose moiety of desmycosin, along with 2‴-O-demethyltylosin (1; 3‴-methyl-demethylmacrocin) that was previously detected from the S. fradiae mutant containing a mutation in the tylE gene. These results unveil the unique substrate flexibility of TylE and TylF and demonstrate their potential for the engineered biosynthesis of novel glycosylated macrolide derivatives.

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Section: Resultssupporting

confidence: 65%

Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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Characterization of the Two Methylation Steps Involved in the Biosynthesis of Mycinose in Tylosin

et al. 2016

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Macrolides

Kirst¹

2000

Kirk-Othmer Encyclopedia of Chemical Technology

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Macrolide antibiotics are well‐established antimicrobial agents in both clinical and veterinary medicine. These agents can be administered orally and are generally used to treat infections in the respiratory tract, skin and soft tissues, and genital tract caused by gram‐positive organisms, Mycoplasma species, and certain susceptible gram‐negative and anaerobic bacteria. The macrolide class is large and structurally diverse. Macrolides are produced by fermentation of soil microorganisms. Additionally, structural modifications using both chemical and microbiological means have yielded biologically active semisynthetic derivatives. The term macrolide was introduced to denote the class of substances produced by Streptomyces species containing a macrocyclic lactone ring. Traditional macrolide antibiotics are divided into three families according to the size of the aglycone, which can be 12‐, 14‐, or 16‐membered. Naturally occurring 14‐membered macrolides include erythromycin A (C 37 H 67 NO 13 ), erythromycin B (C 37 H 67 NO 12 ), erythromycin C (C 36 H 65 NO 13 ), erythromycin D (C 36 H 65 NO 12 ), erythromycin F (C 37 H 67 NO 14 ), and erythromycin E (C 37 H 65 NO 14 ), as well as others. Erythromycin has been the principal subject of modification of 14‐membered macrolides; some of the derivatives of erythromycin and oleandomycin include 2′‐ O ‐acetylerythromycin (C 39 H 69 NO 14 ), 2′‐ O ‐propionylerythromycin (C 40 H 71 NO 14 ), erythromycin ethyl carbonate (C 40 H 71 NO 15 ), and others. 16‐Membered macrolides are divided into leucomycin‐ and tylosin‐related groups, which differ in the substitution pattern of their aglycones. Natural products include leucomycin A 1 (C 40 H 67 NO 14 ), leucomycin A 5 (C 39 H 65 NO 14 ), leucomycin A 7 (C 38 H 63 NO 14 ), midecamycin A 2 (C 42 H 69 NO 15 ), and others. A second large group of 16‐membered macrolides differs from the leucomycins in the substitution pattern of the aglycone. One difference is a methyl or hydroxymethyl group at C‐14. The most prominent member of this group is tylosin, an important veterinary antibiotic produced by S. fradiae. Tylosin and related products include tylosin (C 46 H 77 NO 17 ), relomycin (20‐dihydrotylosin) (C 46 H 79 NO 17 ), macrocin (C 45 H 75 NO 17 ), O ‐demethylmacrocin (C 44 H 73 NO 17 ), and others. Other macrolides have been made by chemical, bioconversion, or genetic manipulations which represent hybrids of structures within the 14‐membered family, within the 16‐membered family, or between the two families. The advent of molecular biology has opened new possibilities for producing hybrid macrolides. Genetic manipulations of biosynthetic pathways in macrolide‐producing microorganisms complement traditional chemical and microbiological approaches. Macrolides inhibit growth of gram‐positive bacteria, Mycoplasma species, and certain gram‐negative and anaerobic bacteria. Susceptible gram‐positive bacteria include many species of Staphylococcus and Streptococcus; susceptible gram‐negative bacteria include Bordetella pertussis, Legionella pneumophila, Moraxella catarrhalis (formerly Branhamella ), and Haemophilus ducreyi. Macrolides inhibit growth of bacteria by inhibiting protein synthesis on ribosomes. Bacterial resistance to macrolides is often accompanied by cross‐resistance to lincosamide and streptogramin B antibiotics (MLS‐resistance). Bacterial resistance to antibiotics usually results from modification of a target site, enzymatic inactivation, or reduced uptake into or increased efflux from bacterial cells. The principal side effects of macrolides are gastrointestinal problems, such as pain, indigestion, diarrhea, nausea, and vomiting. Macrolides are obtained by controlled submerged aerobic fermentations of soil microorganisms. Although species of Streptomyces have dominated, species of Saccharopolyspora, Micromonospora, and Streptoverticillium are also well represented. Macrolide antibiotics are used clinically to treat infections resulting from susceptible organisms in the upper and lower respiratory tract, skin and soft tissues, and genital tract. They are generally used orally, although they can be given intravenously. Macrolides are regarded as among the safest of antibiotics. Relatively few macrolides are used in veterinary medicine. The most important is tylosin (Tylan, Elanco Products), which is used to control chronic respiratory disease caused by Mycoplasma gallisepticum in poultry.

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Antibiotics, Macrolides

Kirst

2001

Kirk-Othmer Encyclopedia of Chemical Technology

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Macrolide antibiotics are well‐established antimicrobial agents in clinical and veterinary medicine. These agents can be administered orally and are generally used to treat infections in the respiratory tract, skin and soft tissues, and genital tract caused by gram‐positive organisms, Mycoplasma species, and certain susceptible gram‐negative and anaerobic bacteria. The macrolide class is large and structurally diverse. Macrolides are produced by fermentation of soil microorganisms. Additionally, structural modifications using both chemical and microbiological means have yielded biologically active semisynthetic derivatives. The term macrolide was introduced to denote the class of substances produced by Streptomyces species containing a macrocyclic lactone. Traditional macrolide antibiotics are divided into three families according to the size of the aglycone, which can be a 12‐, 14‐, or 16‐membered ring. Naturally occurring 14‐membered macrolides include erythromycin A and erythromycin B, as well as others. Erythromycin has been the principal subject of modification of 14‐membered macrolides; its semisynthetic derivatives include erythromycin ethyl succinate, roxithromycin, clarithromycin, azithromycin, dirithromycin, and others. More recently, the ketolide family has been discovered that exhibits activity against certain macrolide‐resistant strains, from which telithromycin and ABT‐773 are in clinical trials. The 16‐membered macrolides are divided into leucomycin‐ and tylosin‐related groups, which differ in the substitution pattern of their aglycones. Natural products include josamycin (leucomycin A 3 ), spiramycin, and others. The most prominent member of the second group is tylosin, an important veterinary antibiotic. Semisynthetic 16‐membered macrolides include miokamycin, rokitamycin, AIV‐tylosin, and tilmicosin. The advent of molecular biology has opened new possibilities for producing novel macrolides by genetic manipulations of biosynthetic pathways from macrolide‐producing microorganisms (combinatorial biosynthesis) that complement traditional chemical and microbiological approaches. Macrolides inhibit growth of gram‐positive bacteria, Mycoplasma species, and certain gram‐negative and anaerobic bacteria. Susceptible gram‐positive bacteria include many species of Staphylococcus and Streptococcus ; susceptible gram‐negative bacteria include Bordetella pertussis, Legionella pneumophila, Moraxella catarrhalis , Campylobacter sp., Chlamydia sp., and Helicobacter pylori . Some derivatives have shown promising activity against non‐tuberculous mycobacteria. Macrolides inhibit growth of bacteria by inhibiting protein synthesis on ribosomes. Bacterial resistance to macrolides is often accompanied by cross‐resistance to lincosamide and streptogramin B antibiotics (MLS‐resistance). Bacterial resistance to antibiotics usually results from modification of a target site, enzymatic inactivation, or reduced uptake into or increased efflux from bacterial cells. The principal side effects of macrolides are gastrointestinal problems, such as pain, indigestion, diarrhea, nausea, and vomiting. Macrolide antibiotics are used clinically to treat infections resulting from susceptible organisms in the upper and lower respiratory tract, skin and soft tissues, and genital tract. They are generally used orally, although they can be given intravenously. Macrolides are regarded as among the safest of antibiotics. A few macrolides are used in veterinary medicine, such as tylosin and tilmicosin which are used to control respiratory diseases and other infections in poultry, pigs, and cattle.

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Two distinctive O-methyltransferases catalyzing penultimate and terminal reactions of macrolide antibiotic (tylosin) biosynthesis. Substrate specificity, enzyme inhibition, and kinetic mechanism.

Cited by 47 publications

References 14 publications

Characterization of the Two Methylation Steps Involved in the Biosynthesis of Mycinose in Tylosin

Characterization of the Two Methylation Steps Involved in the Biosynthesis of Mycinose in Tylosin

Macrolides

Antibiotics, Macrolides

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