Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus

Background: Tannases are enzymes that may be used in different industrial sectors as, for example, food and pharmaceutical. They are obtained mainly from microorganisms, as filamentous fungi. However, the diversity of fungi stays poorly explored for tannase production. In this article, Aspergillus o...

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Autores principales:	Gonçalves,Heloísa Bressan, Riul,Alana Jacomini, Quiapim,Andréa Carla, Jorge,João Atílio, Guimarães,Luis Henrique Souza
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2012
Materias:	Aspergillus ochraceus gallic acid tannase tannic acid tannin acyl hydrolase
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500004
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spelling	oai:scielo:S0717-345820120005000042012-11-29Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceusGonçalves,Heloísa BressanRiul,Alana JacominiQuiapim,Andréa CarlaJorge,João AtílioGuimarães,Luis Henrique Souza Aspergillus ochraceus gallic acid tannase tannic acid tannin acyl hydrolase Background: Tannases are enzymes that may be used in different industrial sectors as, for example, food and pharmaceutical. They are obtained mainly from microorganisms, as filamentous fungi. However, the diversity of fungi stays poorly explored for tannase production. In this article, Aspergillus ochraceus is presented as a new source of tannase with interesting features for biotechnological applications. Results: Extracellular tannase production was induced when the fungus was cultured in Khanna medium with tannic acid as carbon source. The extracellular tannase was purified 9-fold with 2% recovery and a single band corresponding to 85 kDa was observed in SDS-PAGE. The native apparent molecular mass was estimated as 112 kDa. Optima of temperature and pH were 40ºC and 5.0, respectively. The enzyme was fully stable from 40ºC to 60ºC during 1 hr. The activity was enhanced by Mn2+ (33-39%) and NH4+ (15%). The purified tannase hydrolyzed tannic acid and methyl gallate with Km of 0.76 mM and 0.72 mM, respectively, and Vmax of 0.92 U/mg protein and 0.68 U/mg protein, respectively. The analysis of a partial sequence of the tannase encoding gene showed an open read frame of 567 bp and a sequence of 199 amino acids were predicted. TLC analysis revealed the presence of gallic acid as a tannic acid hydrolysis product. Conclusion: The extracellular tannase produced by A. ochraceus showed distinctive characteristics such as monomeric structure and activation by Mn2+, suggesting a new kind of fungal tannases with biotechnological potential. Further, it was the first time that a partial gene sequence for A. ochraceus tannase was described.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.15 n.5 20122012-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500004en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Aspergillus ochraceus gallic acid tannase tannic acid tannin acyl hydrolase
spellingShingle	Aspergillus ochraceus gallic acid tannase tannic acid tannin acyl hydrolase Gonçalves,Heloísa Bressan Riul,Alana Jacomini Quiapim,Andréa Carla Jorge,João Atílio Guimarães,Luis Henrique Souza Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
description	Background: Tannases are enzymes that may be used in different industrial sectors as, for example, food and pharmaceutical. They are obtained mainly from microorganisms, as filamentous fungi. However, the diversity of fungi stays poorly explored for tannase production. In this article, Aspergillus ochraceus is presented as a new source of tannase with interesting features for biotechnological applications. Results: Extracellular tannase production was induced when the fungus was cultured in Khanna medium with tannic acid as carbon source. The extracellular tannase was purified 9-fold with 2% recovery and a single band corresponding to 85 kDa was observed in SDS-PAGE. The native apparent molecular mass was estimated as 112 kDa. Optima of temperature and pH were 40ºC and 5.0, respectively. The enzyme was fully stable from 40ºC to 60ºC during 1 hr. The activity was enhanced by Mn2+ (33-39%) and NH4+ (15%). The purified tannase hydrolyzed tannic acid and methyl gallate with Km of 0.76 mM and 0.72 mM, respectively, and Vmax of 0.92 U/mg protein and 0.68 U/mg protein, respectively. The analysis of a partial sequence of the tannase encoding gene showed an open read frame of 567 bp and a sequence of 199 amino acids were predicted. TLC analysis revealed the presence of gallic acid as a tannic acid hydrolysis product. Conclusion: The extracellular tannase produced by A. ochraceus showed distinctive characteristics such as monomeric structure and activation by Mn2+, suggesting a new kind of fungal tannases with biotechnological potential. Further, it was the first time that a partial gene sequence for A. ochraceus tannase was described.
author	Gonçalves,Heloísa Bressan Riul,Alana Jacomini Quiapim,Andréa Carla Jorge,João Atílio Guimarães,Luis Henrique Souza
author_facet	Gonçalves,Heloísa Bressan Riul,Alana Jacomini Quiapim,Andréa Carla Jorge,João Atílio Guimarães,Luis Henrique Souza
author_sort	Gonçalves,Heloísa Bressan
title	Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
title_short	Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
title_full	Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
title_fullStr	Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
title_full_unstemmed	Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus
title_sort	characterization of a thermostable extracellular tannase produced under submerged fermentation by aspergillus ochraceus
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2012
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000500004
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_version_	1718441860054122496

Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus

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