Inducible nature of the enzymes involved in catabolism of caffeine and related methylxanthines

Dash, Swati Sucharita; Gummadi, Sathyanarayana N.

doi:10.1002/jobm.200800004

Cited by 20 publications

(25 citation statements)

References 20 publications

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“…For instance, when caffeine-degrading Pseudomonas sp. strain NCIM5235 was grown with theobromine as the sole source of carbon and nitrogen, the caffeine N-demethylase and theobromine N-demethylase activities in cell extracts were 10-fold lower and 3-to 4-fold higher, respectively, than those in cell extracts from caffeine-grown cells (2). Caffeine dehydrogenase activity in Pseudomonas sp.…”

Section: Discussionmentioning

confidence: 95%

“…Caffeine and theobromine also did not inhibit 7-methylxanthine N demethylation by this enzyme. A recent study by Dash and Gummadi (2) showed that caffeine and theobromine N-demethylases in Pseudomonas sp. strain NCIM5235 were inducible in nature, but the caffeine N-demethylase activity in theobromine-grown cells was 10-fold lower than that in caffeine-grown cells.…”

Section: Discussionmentioning

confidence: 99%

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Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5

Louie

Summers

et al. 2009

J Bacteriol

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Pseudomonas putida CBB5 was isolated from soil by enrichment on caffeine. This strain used not only caffeine, theobromine, paraxanthine, and 7-methylxanthine as sole carbon and nitrogen sources but also theophylline and 3-methylxanthine. Analyses of metabolites in spent media and resting cell suspensions confirmed that CBB5 initially N demethylated theophylline via a hitherto unreported pathway to 1-and 3-methylxanthines. NAD(P)H-dependent conversion of theophylline to 1-and 3-methylxanthines was also detected in the crude cell extracts of theophylline-grown CBB5. 1-Methylxanthine and 3-methylxanthine were subsequently N demethylated to xanthine. CBB5 also oxidized theophylline and 1-and 3-methylxanthines to 1,3-dimethyluric acid and 1-and 3-methyluric acids, respectively. However, these methyluric acids were not metabolized further. A broad-substrate-range xanthine-oxidizing enzyme was responsible for the formation of these methyluric acids. In contrast, CBB5 metabolized caffeine to theobromine (major metabolite) and paraxanthine (minor metabolite). These dimethylxanthines were further N demethylated to xanthine via 7-methylxanthine. Theobromine-, paraxanthine-, and 7-methylxanthine-grown cells also metabolized all of the methylxanthines mentioned above via the same pathway. Thus, the theophylline and caffeine N-demethylation pathways converged at xanthine via different methylxanthine intermediates. Xanthine was eventually oxidized to uric acid. Enzymes involved in theophylline and caffeine degradation were coexpressed when CBB5 was grown on theophylline or on caffeine or its metabolites. However, 3-methylxanthine-grown CBB5 cells did not metabolize caffeine, whereas theophylline was metabolized at much reduced levels to only methyluric acids. To our knowledge, this is the first report of theophylline N demethylation and coexpression of distinct pathways for caffeine and theophylline degradation in bacteria.Caffeine (1,3,7-trimethylxanthine) and related methylxanthines are widely distributed in many plant species. Caffeine is also a major human dietary ingredient that can be found in common beverages and food products, such as coffee, tea, and chocolates. In pharmaceuticals, caffeine is used generally as a cardiac, neurological, and respiratory stimulant, as well as a diuretic (3). Hence, caffeine and related methylxanthines enter soil and water easily through decomposed plant materials and other means, such as effluents from coffee-and tea-processing facilities. Therefore, it is not surprising that microorganisms capable of degrading caffeine have been isolated from various natural environments, with or without enrichment procedures (3, 10). Bacteria use oxidative and N-demethylating pathways for catabolism of caffeine. Oxidation of caffeine by a Rhodococcus sp.-Klebsiella sp. mixed-culture consortium at the C-8 position to form 1,3,7-trimethyluric acid (TMU) has been reported (8). An 85-kDa, flavin-containing caffeine oxidase was purified from this consortium (9). Also, Mohapatra et al. (12) purified a 65-kDa...

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5

Louie

Summers

et al. 2009

J Bacteriol

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“…Some indirect experiments by others indicate the presence of specific, maybe multiple N-demethylases in caffeine-degrading bacteria. A recent study by Dash & Gummadi (2008) showed that caffeine and theobromine N-demethylases in Pseudomonas sp. NCIM 5235 were inducible in nature but caffeine N-demethylase activity in theobromine-grown cells was 10-fold lower than that of caffeine-grown cells.…”

Section: Discussionmentioning

confidence: 99%

“…The majority of these belong to the genus Pseudomonas (Asano et al, 1993;Blecher & Lingens, 1977;Dash & Gummadi, 2008;Sideso et al, 2001;Woolfolk, 1975;Yu et al, 2009); however, a strain of Serratia marcescens was also found to N-demethylate caffeine (Mazzafera et al, 1996). N-Demethylation of caffeine in bacteria results in production of either theobromine (3,7-dimethylxanthine) or paraxanthine (1,7-dimethylxanthine).…”

Section: Introductionmentioning

confidence: 99%

Characterization of a broad-specificity non-haem iron N-demethylase from Pseudomonas putida CBB5 capable of utilizing several purine alkaloids as sole carbon and nitrogen source

Summers

Louie

et al. 2011

Microbiology

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N-Demethylation of many xenobiotics and naturally occurring purine alkaloids such as caffeine and theobromine is primarily catalysed in higher organisms, ranging from fungi to mammals, by the well-studied membrane-associated cytochrome P450s. In contrast, there is no well-characterized enzyme for N-demethylation of purine alkaloids from bacteria, despite several reports on their utilization as sole source of carbon and nitrogen. Here, we provide what we believe to be the first detailed characterization of a purified N-demethylase from Pseudomonas putida CBB5. The soluble N-demethylase holoenzyme is composed of two components, a reductase component with cytochrome c reductase activity (Ccr) and a two-subunit N-demethylase component (Ndm). Ndm, with a native molecular mass of 240 kDa, is composed of NdmA (40 kDa) and NdmB (35 kDa). Ccr transfers reducing equivalents from NAD(P)H to Ndm, which catalyses an oxygen-dependent N-demethylation of methylxanthines to xanthine, formaldehyde and water. Paraxanthine and 7-methylxanthine were determined to be the best substrates, with apparent K m and kcat values of 50.4±6.8 μM and 16.2±0.6 min−1, and 63.8±7.5 μM and 94.8±3.0 min−1, respectively. Ndm also displayed activity towards caffeine, theobromine, theophylline and 3-methylxanthine, all of which are growth substrates for this organism. Ndm was deduced to be a Rieske [2Fe–2S]-domain-containing non-haem iron oxygenase based on (i) its distinct absorption spectrum and (ii) significant identity of the N-terminal sequences of NdmA and NdmB with the gene product of an uncharacterized caffeine demethylase in P. putida IF-3 and a hypothetical protein in Janthinobacterium sp. Marseille, both predicted to be Rieske non-haem iron oxygenases.

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“…Studies of the enzymes involved in caffeine degradation are also not explicit, with the exception of caffeine oxidase, which has been purified and characterized [24,29]. A few studies have shown there might be separate enzymes, acting separately [1,14] or as a complex [3,12], which bring about specific demethylations in Pseudomonas sp.…”

Section: Introductionmentioning

confidence: 97%

Optimization of production of caffeine demethylase by Pseudomonas sp. in a bioreactor

Gummadi

Dash

Santhosh

2009

J Ind Microbiol Biotechnol

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The effect of pH, aeration rate, and agitation rate on specific productivity of caffeine demethylase from Pseudomonas sp. was studied in a bioreactor. Maximum specific productivity of caffeine demethylase of 2,214 U g cell dry weight(-1) h(-1) was obtained at 0.27 vvm, 700 rpm, and pH 7.0. Under these conditions, volumetric oxygen transfer coefficient was 74.2 h(-1), indicating that caffeine demethylase production by Pseudomonas sp. was highly oxygen-dependent. Different metabolite formation at different agitation and aeration rates can be used as a strategy for recovery of pharmaceutically important metabolites from caffeine by manipulation of conditions in a bacterial culture. This is the first report on production of high levels of caffeine demethylase in bioreactors.

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Inducible nature of the enzymes involved in catabolism of caffeine and related methylxanthines

Cited by 20 publications

References 20 publications

Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5

Two Distinct Pathways for Metabolism of Theophylline and Caffeine Are Coexpressed in Pseudomonas putida CBB5

Characterization of a broad-specificity non-haem iron N-demethylase from Pseudomonas putida CBB5 capable of utilizing several purine alkaloids as sole carbon and nitrogen source

Optimization of production of caffeine demethylase by Pseudomonas sp. in a bioreactor

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