Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1080/10826068.2015.1031387 http://hdl.handle.net/11449/158832 |
Resumo: | Peptidases are important because they play a central role in pharmaceutical, food, environmental, and other industrial processes. A serine peptidase from Aspergillus terreus was isolated after two chromatography steps that showed a yield of 15.5%. Its molecular mass was determined to be 43 kD, by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This peptidase was active between pH 5.0 to 8.0 and had maximum activity at pH 7.0, at 45 degrees C. When exposited with 1 M of urea, the enzyme maintained 100% activity and used azocasein as substrate. The N-terminal (first 15 residues) showed 33% identity with the serine peptidase of Aspergillus clavatus ES1. The kinetics assays showed that subsite S-2 did not bind polar basic amino acids (His and Arg) nonpolar acidic amino acids (Asp and Glu). The subsite S-1 showed higher catalytic efficiency than the S-2 and S-3 subsites. |
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Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreusAspergillusenzyme kineticsfilamentous fungiN-terminal sequenceproteaseproteinPeptidases are important because they play a central role in pharmaceutical, food, environmental, and other industrial processes. A serine peptidase from Aspergillus terreus was isolated after two chromatography steps that showed a yield of 15.5%. Its molecular mass was determined to be 43 kD, by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This peptidase was active between pH 5.0 to 8.0 and had maximum activity at pH 7.0, at 45 degrees C. When exposited with 1 M of urea, the enzyme maintained 100% activity and used azocasein as substrate. The N-terminal (first 15 residues) showed 33% identity with the serine peptidase of Aspergillus clavatus ES1. The kinetics assays showed that subsite S-2 did not bind polar basic amino acids (His and Arg) nonpolar acidic amino acids (Asp and Glu). The subsite S-1 showed higher catalytic efficiency than the S-2 and S-3 subsites.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Sao Paulo, Fac Pharmaceut Sci Ribeirao Preto, Dept Pharmaceut Sci, S-N Cafe, BR-14040903 Ribeirao Preto, BrazilUniv Estadual Paulista, Inst Biosci Letters & Exact Sci, Sao Jose Do Rio Preto, BrazilFed Univ Grande Dourados, Fac Biol & Environm Sci, Dourados, BrazilUniv Sao Paulo, Fac Pharmaceut Sci Ribeirao Preto, Dept Chem & Phys, BR-14040903 Ribeirao Preto, BrazilUniv Ctr Educ Fdn Barretos, Fac Pharm, Barretos, BrazilUniv Fed Sao Paulo, Paulista Sch Med, Dept Biophys, Sao Paulo, BrazilUniv Estadual Paulista, Inst Biosci Letters & Exact Sci, Sao Jose Do Rio Preto, BrazilFAPESP: 2011/06986-0FAPESP: 2012/24703-8CNPq: 308078/2012-8Taylor & Francis IncUniversidade de São Paulo (USP)Universidade Estadual Paulista (Unesp)Fed Univ Grande DouradosUniv Ctr Educ Fdn BarretosUniversidade Federal de São Paulo (UNIFESP)Biaggio, Rafael TageSilva, Ronivaldo Rodrigues da [UNESP]Rosa, Nathalia Gonsales daRibeiro Leite, Rodrigo SimoesArantes, Eliane CandianiFreitas Cabral, Tatiana Pereira deJuliano, Maria A.Juliano, LuizCabral, Hamilton2018-11-26T15:29:22Z2018-11-26T15:29:22Z2016-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article298-304application/pdfhttp://dx.doi.org/10.1080/10826068.2015.1031387Preparative Biochemistry & Biotechnology. Philadelphia: Taylor & Francis Inc, v. 46, n. 3, p. 298-304, 2016.1082-6068http://hdl.handle.net/11449/15883210.1080/10826068.2015.1031387WOS:000374887000012WOS000374887000012.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPreparative Biochemistry & Biotechnology0,362info:eu-repo/semantics/openAccess2024-01-07T06:25:54Zoai:repositorio.unesp.br:11449/158832Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T22:20:55.636867Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
title |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
spellingShingle |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus Biaggio, Rafael Tage Aspergillus enzyme kinetics filamentous fungi N-terminal sequence protease protein |
title_short |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
title_full |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
title_fullStr |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
title_full_unstemmed |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
title_sort |
Purification and biochemical characterization of an extracellular serine peptidase from Aspergillus terreus |
author |
Biaggio, Rafael Tage |
author_facet |
Biaggio, Rafael Tage Silva, Ronivaldo Rodrigues da [UNESP] Rosa, Nathalia Gonsales da Ribeiro Leite, Rodrigo Simoes Arantes, Eliane Candiani Freitas Cabral, Tatiana Pereira de Juliano, Maria A. Juliano, Luiz Cabral, Hamilton |
author_role |
author |
author2 |
Silva, Ronivaldo Rodrigues da [UNESP] Rosa, Nathalia Gonsales da Ribeiro Leite, Rodrigo Simoes Arantes, Eliane Candiani Freitas Cabral, Tatiana Pereira de Juliano, Maria A. Juliano, Luiz Cabral, Hamilton |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Estadual Paulista (Unesp) Fed Univ Grande Dourados Univ Ctr Educ Fdn Barretos Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Biaggio, Rafael Tage Silva, Ronivaldo Rodrigues da [UNESP] Rosa, Nathalia Gonsales da Ribeiro Leite, Rodrigo Simoes Arantes, Eliane Candiani Freitas Cabral, Tatiana Pereira de Juliano, Maria A. Juliano, Luiz Cabral, Hamilton |
dc.subject.por.fl_str_mv |
Aspergillus enzyme kinetics filamentous fungi N-terminal sequence protease protein |
topic |
Aspergillus enzyme kinetics filamentous fungi N-terminal sequence protease protein |
description |
Peptidases are important because they play a central role in pharmaceutical, food, environmental, and other industrial processes. A serine peptidase from Aspergillus terreus was isolated after two chromatography steps that showed a yield of 15.5%. Its molecular mass was determined to be 43 kD, by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This peptidase was active between pH 5.0 to 8.0 and had maximum activity at pH 7.0, at 45 degrees C. When exposited with 1 M of urea, the enzyme maintained 100% activity and used azocasein as substrate. The N-terminal (first 15 residues) showed 33% identity with the serine peptidase of Aspergillus clavatus ES1. The kinetics assays showed that subsite S-2 did not bind polar basic amino acids (His and Arg) nonpolar acidic amino acids (Asp and Glu). The subsite S-1 showed higher catalytic efficiency than the S-2 and S-3 subsites. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-01-01 2018-11-26T15:29:22Z 2018-11-26T15:29:22Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1080/10826068.2015.1031387 Preparative Biochemistry & Biotechnology. Philadelphia: Taylor & Francis Inc, v. 46, n. 3, p. 298-304, 2016. 1082-6068 http://hdl.handle.net/11449/158832 10.1080/10826068.2015.1031387 WOS:000374887000012 WOS000374887000012.pdf |
url |
http://dx.doi.org/10.1080/10826068.2015.1031387 http://hdl.handle.net/11449/158832 |
identifier_str_mv |
Preparative Biochemistry & Biotechnology. Philadelphia: Taylor & Francis Inc, v. 46, n. 3, p. 298-304, 2016. 1082-6068 10.1080/10826068.2015.1031387 WOS:000374887000012 WOS000374887000012.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Preparative Biochemistry & Biotechnology 0,362 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
298-304 application/pdf |
dc.publisher.none.fl_str_mv |
Taylor & Francis Inc |
publisher.none.fl_str_mv |
Taylor & Francis Inc |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1808129418343743488 |