Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42)
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tde/1267 |
Resumo: | Trichoderma harzianum is a saprophytic fungus, known for its potential as a biological control agent of different phytopathogens that causes losses in crops. Its action is based on different mechanisms like volatile and non-volatile antibiotics production, competition for nutrient and space, production of hydrolytic enzymes and mycoparasitism. This fungus also plays an important role in the release of carbon, nitrogen and phosphorus from insoluble macromolecules to the medium, favoring the growth of plants. Phosphorus is a limiting nutrient for plant growth, however, over 80% of the phosphorus applied to the soil, it becomes unavailable, due to its adsorption, precipitation or conversion to organic form. One way to obtain phosphate compounds in soil is through the action of enzymes called phosphatases, which catalyze the hydrolysis of phosphate esters producing soluble phosphorus. High levels of acid phosphatase (ACP) were produced by Trichoderma harzianum ALL42. This study evaluated the ability of T. harzianumALL42produce acid phosphatases(ACPs) in minimal medium modified by varying the concentration of glucose and phosphate (KH2PO4). The results showed that the concentration of glucose and phosphate in the culture medium regulated the production of ACPs T. harzianum ALL42. Thisfungusproducedanacid phosphatase(ACPII) inculture mediumcontainingglucose0.5% and0.04% phosphate. Theenzymewaspartiallypurifiedby hydrophobic interaction chromatography on Phenyl Sepharose. A typical procedure provided 2,0 fold purification with 32.65 % yield. It was optimally active in the pH range 5,2 and at 50°C. The enzyme was heat-stable, retaining approximately 60% of its activity after heating for 60 min at 60°C. Kinetic parameters calculated for the hydrolysis of p-nitrophenyl phosphate by acid phosphatase were Km= 0,054 M and Vmax= 2,058 units.min-1. The enzyme was strongly inhibited by KH2PO4, FeCl3 and sodium tungstate. The enzyme is inhibited by KH2PO4 and sodium tungstate through mixed inhibition. The acid phosphatase hydrolyzed a number of phosphate esters ATP, ADP, AMP, D-glucose-1-phosphate, D-fructose-6-phosphate, includingphytic acid, showinganactivityofphytase. |
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ULHOA, Cirano Josehttp://lattes.cnpq.br/8368469162867277FREITAS, Sonia Maria dehttp://lattes.cnpq.br/8369464483722617http://lattes.cnpq.br/5791449510340723SOUZA, Amanda Araujo2014-07-29T15:16:32Z2012-11-202011-06-29SOUZA, Amanda Araujo. Production and biochemistry caracterization of the acid phosphatase Trichoderma harzianum (ALL42). 2011. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2011.http://repositorio.bc.ufg.br/tede/handle/tde/1267Trichoderma harzianum is a saprophytic fungus, known for its potential as a biological control agent of different phytopathogens that causes losses in crops. Its action is based on different mechanisms like volatile and non-volatile antibiotics production, competition for nutrient and space, production of hydrolytic enzymes and mycoparasitism. This fungus also plays an important role in the release of carbon, nitrogen and phosphorus from insoluble macromolecules to the medium, favoring the growth of plants. Phosphorus is a limiting nutrient for plant growth, however, over 80% of the phosphorus applied to the soil, it becomes unavailable, due to its adsorption, precipitation or conversion to organic form. One way to obtain phosphate compounds in soil is through the action of enzymes called phosphatases, which catalyze the hydrolysis of phosphate esters producing soluble phosphorus. High levels of acid phosphatase (ACP) were produced by Trichoderma harzianum ALL42. This study evaluated the ability of T. harzianumALL42produce acid phosphatases(ACPs) in minimal medium modified by varying the concentration of glucose and phosphate (KH2PO4). The results showed that the concentration of glucose and phosphate in the culture medium regulated the production of ACPs T. harzianum ALL42. Thisfungusproducedanacid phosphatase(ACPII) inculture mediumcontainingglucose0.5% and0.04% phosphate. Theenzymewaspartiallypurifiedby hydrophobic interaction chromatography on Phenyl Sepharose. A typical procedure provided 2,0 fold purification with 32.65 % yield. It was optimally active in the pH range 5,2 and at 50°C. The enzyme was heat-stable, retaining approximately 60% of its activity after heating for 60 min at 60°C. Kinetic parameters calculated for the hydrolysis of p-nitrophenyl phosphate by acid phosphatase were Km= 0,054 M and Vmax= 2,058 units.min-1. The enzyme was strongly inhibited by KH2PO4, FeCl3 and sodium tungstate. The enzyme is inhibited by KH2PO4 and sodium tungstate through mixed inhibition. The acid phosphatase hydrolyzed a number of phosphate esters ATP, ADP, AMP, D-glucose-1-phosphate, D-fructose-6-phosphate, includingphytic acid, showinganactivityofphytase.Trichoderma harzianum é um fungo saprofítico, conhecido pelo seu grande potencial como agente de controle biológico de diferentes fitopatógenos. Sua ação é baseada em diferentes mecanismos como a produção de antibióticos voláteis e não-voláteis, competição por espaço e nutrientes, produção de enzimas hidrolíticas e o micoparasitismo. Esse fungo também desempenha um importante papel na liberação de carbono, nitrogênio e fósforo, a partir de macromoléculas insolúveis, para o meio, favorecendo o crescimento de plantas. O fósforo é um nutriente limitante para o crescimento da planta, entretanto, mais de 80% do fósforo aplicado diretamente no solo, torna-se indisponível, devido a sua adsorção, precipitação ou conversão para forma orgânica. Uma forma de se obter compostos fosfatados no solo é através da ação de enzimas denominadas fosfatases, que catalisam a hidrólise de ésteres fosfatados produzindo fósforo solúvel. Altos níveis de fosfatase ácida foram produzidos por Trichoderma harzianum ALL42. Neste trabalho avaliou-se a capacidade de T. harzianum ALL42 produzir fosfatases ácidas (ACPs) em meio mínimo modificado, variando a concentração de glicose e fosfato (KH2PO4). Os resultados obtidos demonstraram que, a concentração de glicose e fosfato no meio de cultura regulou a produção de ACPs por T. harzianum ALL42. Esse fungoproduziu uma fosfatase ácida (ACPII) em meio de cultura contendo glicose 0,5% e fosfato 0,04%. A enzima foi parcialmente purificada através da cromatografia de interação hidrofóbica Phenyl-Sepharose. O processo de purificação obteve um fator de purificação de 2,0 e rendimento de 32,65%. A atividade ótima encontrada foi na faixa de pH 5,2 e a 50 ºC. A enzima foi termoestável, mantendo aproximadamente 60% de sua atividade após incubação por 60 min a 60 ºC. Os parâmetros cinéticos calculados pela hidrólise de ƿ-nitrofenilfosfato pela fosfatase ácida foram de Km = 0,054 µmol e Vmáx = 2,028U.min-1. A enzima foi fortemente inibida por KH2PO4,FeCl3 e tungstato de sódio. A enzima foi inibida por KH2PO4 e tungstato de sódio por inibição mista. A fosfatase ácida hidrolisou todos os ésteres fosfatados testados (ATP, ADP, AMP, D-glicose-1-fosfato, D-frutose-6-fosfato, fenil fosfato de sódio), inclusive ácido fítico, demonstrando uma atividade de fitase.Made available in DSpace on 2014-07-29T15:16:32Z (GMT). No. of bitstreams: 1 dissertacao de mestrado completa Amanda A Souza.pdf: 994299 bytes, checksum: 8cf098dd70299abaf08a216436f38888 (MD5) Previous issue date: 2011-06-29application/pdfhttp://repositorio.bc.ufg.br/TEDE/retrieve/3840/dissertacao%20%20de%20mestrado%20completa%20Amanda%20A%20Souza.pdf.jpgporUniversidade Federal de GoiásMestrado em BiologiaUFGBRCiências BiolóicasTrichoderma harzianumsolubilização de fosfatofosfatase ácidaTrichoderma harzianum, phosphate solubilization, acid phosphataseCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIAProdução e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42)Production and biochemistry caracterization of the acid phosphatase Trichoderma harzianum (ALL42)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGORIGINALdissertacao de mestrado completa Amanda A Souza.pdfapplication/pdf994299http://repositorio.bc.ufg.br/tede/bitstreams/9bb94542-1a45-45d6-996b-ff1a34856625/download8cf098dd70299abaf08a216436f38888MD51THUMBNAILdissertacao de mestrado completa Amanda A Souza.pdf.jpgdissertacao de mestrado completa Amanda A Souza.pdf.jpgGenerated Thumbnailimage/jpeg2132http://repositorio.bc.ufg.br/tede/bitstreams/dba12bf8-deb0-43ae-9799-4941fb1e7f05/download9991e27c67fca1c59009d80839bc12ceMD52tde/12672014-07-30 03:08:39.964open.accessoai:repositorio.bc.ufg.br:tde/1267http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2014-07-30T06:08:39Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)false |
dc.title.por.fl_str_mv |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
dc.title.alternative.eng.fl_str_mv |
Production and biochemistry caracterization of the acid phosphatase Trichoderma harzianum (ALL42) |
title |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
spellingShingle |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) SOUZA, Amanda Araujo Trichoderma harzianum solubilização de fosfato fosfatase ácida Trichoderma harzianum, phosphate solubilization, acid phosphatase CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
title_short |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
title_full |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
title_fullStr |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
title_full_unstemmed |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
title_sort |
Produção e caracterização bioquímica de uma fosfatase ácida de Trichoderma harzianum (ALL42) |
author |
SOUZA, Amanda Araujo |
author_facet |
SOUZA, Amanda Araujo |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
ULHOA, Cirano Jose |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8368469162867277 |
dc.contributor.advisor-co1.fl_str_mv |
FREITAS, Sonia Maria de |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/8369464483722617 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5791449510340723 |
dc.contributor.author.fl_str_mv |
SOUZA, Amanda Araujo |
contributor_str_mv |
ULHOA, Cirano Jose FREITAS, Sonia Maria de |
dc.subject.por.fl_str_mv |
Trichoderma harzianum solubilização de fosfato fosfatase ácida |
topic |
Trichoderma harzianum solubilização de fosfato fosfatase ácida Trichoderma harzianum, phosphate solubilization, acid phosphatase CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
dc.subject.eng.fl_str_mv |
Trichoderma harzianum, phosphate solubilization, acid phosphatase |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::ENZIMOLOGIA |
description |
Trichoderma harzianum is a saprophytic fungus, known for its potential as a biological control agent of different phytopathogens that causes losses in crops. Its action is based on different mechanisms like volatile and non-volatile antibiotics production, competition for nutrient and space, production of hydrolytic enzymes and mycoparasitism. This fungus also plays an important role in the release of carbon, nitrogen and phosphorus from insoluble macromolecules to the medium, favoring the growth of plants. Phosphorus is a limiting nutrient for plant growth, however, over 80% of the phosphorus applied to the soil, it becomes unavailable, due to its adsorption, precipitation or conversion to organic form. One way to obtain phosphate compounds in soil is through the action of enzymes called phosphatases, which catalyze the hydrolysis of phosphate esters producing soluble phosphorus. High levels of acid phosphatase (ACP) were produced by Trichoderma harzianum ALL42. This study evaluated the ability of T. harzianumALL42produce acid phosphatases(ACPs) in minimal medium modified by varying the concentration of glucose and phosphate (KH2PO4). The results showed that the concentration of glucose and phosphate in the culture medium regulated the production of ACPs T. harzianum ALL42. Thisfungusproducedanacid phosphatase(ACPII) inculture mediumcontainingglucose0.5% and0.04% phosphate. Theenzymewaspartiallypurifiedby hydrophobic interaction chromatography on Phenyl Sepharose. A typical procedure provided 2,0 fold purification with 32.65 % yield. It was optimally active in the pH range 5,2 and at 50°C. The enzyme was heat-stable, retaining approximately 60% of its activity after heating for 60 min at 60°C. Kinetic parameters calculated for the hydrolysis of p-nitrophenyl phosphate by acid phosphatase were Km= 0,054 M and Vmax= 2,058 units.min-1. The enzyme was strongly inhibited by KH2PO4, FeCl3 and sodium tungstate. The enzyme is inhibited by KH2PO4 and sodium tungstate through mixed inhibition. The acid phosphatase hydrolyzed a number of phosphate esters ATP, ADP, AMP, D-glucose-1-phosphate, D-fructose-6-phosphate, includingphytic acid, showinganactivityofphytase. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011-06-29 |
dc.date.available.fl_str_mv |
2012-11-20 |
dc.date.accessioned.fl_str_mv |
2014-07-29T15:16:32Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
SOUZA, Amanda Araujo. Production and biochemistry caracterization of the acid phosphatase Trichoderma harzianum (ALL42). 2011. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2011. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tde/1267 |
identifier_str_mv |
SOUZA, Amanda Araujo. Production and biochemistry caracterization of the acid phosphatase Trichoderma harzianum (ALL42). 2011. 82 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2011. |
url |
http://repositorio.bc.ufg.br/tede/handle/tde/1267 |
dc.language.iso.fl_str_mv |
por |
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por |
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openAccess |
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Universidade Federal de Goiás |
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Mestrado em Biologia |
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UFG |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Ciências Biolóicas |
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Universidade Federal de Goiás |
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