Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L
Autor(a) principal: | |
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Data de Publicação: | 1999 |
Outros Autores: | |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista de Microbiologia |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000100013 |
Resumo: | Caffeine catabolism and a xanthine oxidase involved in the alkaloid breakdown were studied in Pseudomonas putida L, a strain displaying high ability to grow on this substrate. Cells cultured with unlabelled caffeine and 14C labeled caffeine and xanthine showed that this alkaloid was broken-down via theobromine/paraxanthine -> 7-methylxanthine -> xanthine -> uric acid -> allantoin -> allantoic acid. Methyluric acids were formed from the oxidation of theobromine, paraxanthine and 7-methylxanthine, although no bacterial growth was observed on these compounds, indicating that this might be due to a wide substrate specificity of xanthine oxidase. This was confirmed by activity staining in PAGE where activity was observed with theophylline and 3-methylxanthine, which are not involved in the alkaloid breakdown. A single band of activity was detected without addition of NAD+, showing an oxidase form of the enzyme. The enzyme optimum temperature and pH were 30oC and 7.0, respectively. The determined Km was 169 µM, and the pI 3.1 - 4.0. The molecular weight determined by side by side comparison of activity staining of the enzyme in PAGE and PAGE of BSA was 192 kDa, which was coincident with the sum (198.4 kDa) of three subunits (71, 65.6 and 61.8 kDa) of the purified protein. |
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Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida Lcaffeinemethylpurinesmethyluric acidsPseudomonasxanthine oxidaseCaffeine catabolism and a xanthine oxidase involved in the alkaloid breakdown were studied in Pseudomonas putida L, a strain displaying high ability to grow on this substrate. Cells cultured with unlabelled caffeine and 14C labeled caffeine and xanthine showed that this alkaloid was broken-down via theobromine/paraxanthine -> 7-methylxanthine -> xanthine -> uric acid -> allantoin -> allantoic acid. Methyluric acids were formed from the oxidation of theobromine, paraxanthine and 7-methylxanthine, although no bacterial growth was observed on these compounds, indicating that this might be due to a wide substrate specificity of xanthine oxidase. This was confirmed by activity staining in PAGE where activity was observed with theophylline and 3-methylxanthine, which are not involved in the alkaloid breakdown. A single band of activity was detected without addition of NAD+, showing an oxidase form of the enzyme. The enzyme optimum temperature and pH were 30oC and 7.0, respectively. The determined Km was 169 µM, and the pI 3.1 - 4.0. The molecular weight determined by side by side comparison of activity staining of the enzyme in PAGE and PAGE of BSA was 192 kDa, which was coincident with the sum (198.4 kDa) of three subunits (71, 65.6 and 61.8 kDa) of the purified protein.Sociedade Brasileira de Microbiologia1999-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000100013Revista de Microbiologia v.30 n.1 1999reponame:Revista de Microbiologiainstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S0001-37141999000100013info:eu-repo/semantics/openAccessYamaoka-Yano,Dirce MithicoMazzafera,Pauloeng1999-09-17T00:00:00Zoai:scielo:S0001-37141999000100013Revistahttps://www.scielo.br/j/rm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||revmicro@icb.usp.br0001-37140001-3714opendoar:1999-09-17T00:00Revista de Microbiologia - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
title |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
spellingShingle |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L Yamaoka-Yano,Dirce Mithico caffeine methylpurines methyluric acids Pseudomonas xanthine oxidase |
title_short |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
title_full |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
title_fullStr |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
title_full_unstemmed |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
title_sort |
Catabolism of caffeine and purification of a xanthine oxidade responsible for methyluric acids production in Pseudomonas putida L |
author |
Yamaoka-Yano,Dirce Mithico |
author_facet |
Yamaoka-Yano,Dirce Mithico Mazzafera,Paulo |
author_role |
author |
author2 |
Mazzafera,Paulo |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Yamaoka-Yano,Dirce Mithico Mazzafera,Paulo |
dc.subject.por.fl_str_mv |
caffeine methylpurines methyluric acids Pseudomonas xanthine oxidase |
topic |
caffeine methylpurines methyluric acids Pseudomonas xanthine oxidase |
description |
Caffeine catabolism and a xanthine oxidase involved in the alkaloid breakdown were studied in Pseudomonas putida L, a strain displaying high ability to grow on this substrate. Cells cultured with unlabelled caffeine and 14C labeled caffeine and xanthine showed that this alkaloid was broken-down via theobromine/paraxanthine -> 7-methylxanthine -> xanthine -> uric acid -> allantoin -> allantoic acid. Methyluric acids were formed from the oxidation of theobromine, paraxanthine and 7-methylxanthine, although no bacterial growth was observed on these compounds, indicating that this might be due to a wide substrate specificity of xanthine oxidase. This was confirmed by activity staining in PAGE where activity was observed with theophylline and 3-methylxanthine, which are not involved in the alkaloid breakdown. A single band of activity was detected without addition of NAD+, showing an oxidase form of the enzyme. The enzyme optimum temperature and pH were 30oC and 7.0, respectively. The determined Km was 169 µM, and the pI 3.1 - 4.0. The molecular weight determined by side by side comparison of activity staining of the enzyme in PAGE and PAGE of BSA was 192 kDa, which was coincident with the sum (198.4 kDa) of three subunits (71, 65.6 and 61.8 kDa) of the purified protein. |
publishDate |
1999 |
dc.date.none.fl_str_mv |
1999-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000100013 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000100013 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0001-37141999000100013 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Revista de Microbiologia v.30 n.1 1999 reponame:Revista de Microbiologia instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Revista de Microbiologia |
collection |
Revista de Microbiologia |
repository.name.fl_str_mv |
Revista de Microbiologia - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||revmicro@icb.usp.br |
_version_ |
1754821030175571968 |