Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta
Extracellular proteases from Brevundimonas diminuta (syn. Pseudomonas diminuta) were studied in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) containing a copolymerized substrate. Two proteases were detected migrating at 67 kDa and 50 kDa: both of them hydrolysed preferentiall...
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Sociedade Brasileira de Microbiologia
2000-03-01
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doaj-307d3017c62e461982388e43919c03bd2020-11-24T23:33:53ZengSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology1517-83821678-44052000-03-01311252910.1590/S1517-83822000000100007Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminutaAndré Adriano ChaiaSalvatore Giovanni-De-SimoneSimone Dias Gonçalves PetinateAna Paula Cabral de Araújo LimaMarta Helena BranquinhaAlane Beatriz VermelhoExtracellular proteases from Brevundimonas diminuta (syn. Pseudomonas diminuta) were studied in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) containing a copolymerized substrate. Two proteases were detected migrating at 67 kDa and 50 kDa: both of them hydrolysed preferentially gelatin, but casein was also degraded and a slight hydrolysis was observed with hemoglobin. No detectable extracellular proteolytic activity was found in bovine serum albumin-containing gels. The optima temperature and pH for proteolytic activity were between 40ºC and 50ºC in a pH ranging from 7.0 to 11.0, respectively. These enzymes were isolated by analytical high performance liquid chromatography (HPLC). Protease assays with the synthetic substrate Z-Phe-Arg-MCA and the inhibitors EGTA, EDTA and 1, 10 phenanthroline point out that these enzymes are metalloproteases.<br>Proteases extracelulares de Brevundimonas diminuta (Pseudomonas diminuta) foram identificadas e caracterizadas por eletroforese em gel de poliacrilamida com dodecilsulfato de sódio, contendo um substrato co-polimerizado. Duas proteases foram detectadas migrando em 67 kDa e 50 kDa: ambas hidrolisaram preferencialmente a gelatina, embora a caseína também tenha sido degradada e uma pequena hidrólise tenha sido observada com hemoglobina. Nenhuma atividade proteolítica extracelular foi detectada nos géis contendo soro albumina bovina. Condições ótimas de temperatura e pH para a atividade proteolítica foram observadas entre 40ºC e 50ºC e numa faixa de pH que variou de 7,0 a 11,0, respectivamente. Essas enzimas foram isoladas por cromatografia líquida de alta resolução. Os ensaios enzimáticos com o substrato sintético Z-Phe-Arg-MCA e com os inibidores EGTA, EDTA e 1, 10 fenantrolina indicam que essas enzimas são metaloproteases.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822000000100007metaloproteasesBrevundimonas diminutaPseudomonadaceaeproteases extracelularesmetalloproteasesBrevundimonas diminutaPseudomonadaceaeextracellular proteases |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
André Adriano Chaia Salvatore Giovanni-De-Simone Simone Dias Gonçalves Petinate Ana Paula Cabral de Araújo Lima Marta Helena Branquinha Alane Beatriz Vermelho |
spellingShingle |
André Adriano Chaia Salvatore Giovanni-De-Simone Simone Dias Gonçalves Petinate Ana Paula Cabral de Araújo Lima Marta Helena Branquinha Alane Beatriz Vermelho Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta Brazilian Journal of Microbiology metaloproteases Brevundimonas diminuta Pseudomonadaceae proteases extracelulares metalloproteases Brevundimonas diminuta Pseudomonadaceae extracellular proteases |
author_facet |
André Adriano Chaia Salvatore Giovanni-De-Simone Simone Dias Gonçalves Petinate Ana Paula Cabral de Araújo Lima Marta Helena Branquinha Alane Beatriz Vermelho |
author_sort |
André Adriano Chaia |
title |
Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta |
title_short |
Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta |
title_full |
Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta |
title_fullStr |
Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta |
title_full_unstemmed |
Identification and properties of two extracellular proteases from Brevundimonas diminuta Identificação e propriedades de duas proteases extracelulares de Brevundimonas diminuta |
title_sort |
identification and properties of two extracellular proteases from brevundimonas diminuta identificação e propriedades de duas proteases extracelulares de brevundimonas diminuta |
publisher |
Sociedade Brasileira de Microbiologia |
series |
Brazilian Journal of Microbiology |
issn |
1517-8382 1678-4405 |
publishDate |
2000-03-01 |
description |
Extracellular proteases from Brevundimonas diminuta (syn. Pseudomonas diminuta) were studied in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) containing a copolymerized substrate. Two proteases were detected migrating at 67 kDa and 50 kDa: both of them hydrolysed preferentially gelatin, but casein was also degraded and a slight hydrolysis was observed with hemoglobin. No detectable extracellular proteolytic activity was found in bovine serum albumin-containing gels. The optima temperature and pH for proteolytic activity were between 40ºC and 50ºC in a pH ranging from 7.0 to 11.0, respectively. These enzymes were isolated by analytical high performance liquid chromatography (HPLC). Protease assays with the synthetic substrate Z-Phe-Arg-MCA and the inhibitors EGTA, EDTA and 1, 10 phenanthroline point out that these enzymes are metalloproteases.<br>Proteases extracelulares de Brevundimonas diminuta (Pseudomonas diminuta) foram identificadas e caracterizadas por eletroforese em gel de poliacrilamida com dodecilsulfato de sódio, contendo um substrato co-polimerizado. Duas proteases foram detectadas migrando em 67 kDa e 50 kDa: ambas hidrolisaram preferencialmente a gelatina, embora a caseína também tenha sido degradada e uma pequena hidrólise tenha sido observada com hemoglobina. Nenhuma atividade proteolítica extracelular foi detectada nos géis contendo soro albumina bovina. Condições ótimas de temperatura e pH para a atividade proteolítica foram observadas entre 40ºC e 50ºC e numa faixa de pH que variou de 7,0 a 11,0, respectivamente. Essas enzimas foram isoladas por cromatografia líquida de alta resolução. Os ensaios enzimáticos com o substrato sintético Z-Phe-Arg-MCA e com os inibidores EGTA, EDTA e 1, 10 fenantrolina indicam que essas enzimas são metaloproteases. |
topic |
metaloproteases Brevundimonas diminuta Pseudomonadaceae proteases extracelulares metalloproteases Brevundimonas diminuta Pseudomonadaceae extracellular proteases |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822000000100007 |
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