Purifica????o e caracteriza????o da lacase do fungo Trametes cubensis (Mont.) Sacc. 1891

Autor: Damasceno, Andrey Azedo
Přispěvatelé: Campos, Ceci Sales, Silva, Ademir Castro e
Jazyk: portugalština
Rok vydání: 2016
Předmět:
Zdroj: Biblioteca Digital de Teses e Dissertações da UFAM
Universidade Federal do Amazonas (UFAM)
instacron:UFAM
Popis: Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-07-31T15:38:22Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Andrey A. Damasceno.pdf: 2116345 bytes, checksum: 7c6f07c8d1a70353d64b35efff86a951 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-07-31T15:38:34Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Andrey A. Damasceno.pdf: 2116345 bytes, checksum: 7c6f07c8d1a70353d64b35efff86a951 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-07-31T15:38:57Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Andrey A. Damasceno.pdf: 2116345 bytes, checksum: 7c6f07c8d1a70353d64b35efff86a951 (MD5) Made available in DSpace on 2017-07-31T15:38:57Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Andrey A. Damasceno.pdf: 2116345 bytes, checksum: 7c6f07c8d1a70353d64b35efff86a951 (MD5) Previous issue date: 2016-05-27 CNPq - Conselho Nacional de Desenvolvimento Cient??fico e Tecnol??gico The following interest in fungal enzymes comes about their potential for industrial use, as well as biotechnological research. In this context, the enzyme laccase (EC 1.10.3.2) stand out by its potential use in several industrial applications. Thus, the main goal of this research was to characterize the laccase enzyme produced by the Amazonian fungus Trametes cubensis. The T. cubensis strain was collected in Tef?? National Forest (FLONA) and the enzymatic activity was determined from the submerged and semi-solid fermentation. As substrate, sawdust of the wood of Simarouba amara and sugarcane bagasse was tested. As inductors, copper sulfate, ???ABTS??? and mineral medium were used. The laccase was purified by means of ion exchange chromotography and characterized by mass (SDS-PAGE and zymogram) and determined kinetic parameters Km and Vmax. The most expressive laccase activity occurred in the medium with substrate of Simarouba amara (marup??) with 184 U / mL. The yield was 40% and purification factor of 1.5, with Vmax and Km of 33.33 ??mol.L-1 6.66 mM, respectively. The becoming laccase from Trametes cubensis has an apparent molecular mass of approximately 40 kDa. In this way, from the cultivation of Trametes cubensis, it is possible to obtain laccases, however, the study is still necessary in reference the use of different inductors, looking for a potentialization of this activity. O crescente interesse pelas enzimas f??ngicas vem em fun????o da sua potencialidade para uso industrial, bem como na pesquisa biotecnol??gica. Neste contexto, a enzima lacase (EC 1.10.3.2) destaca-se pelo seu potencial uso em diversas aplicabilidades industriais. Assim, o objetivo dessa pesquisa foi caracterizar a enzima lacase produzida pelo fungo amaz??nico Trametes cubensis. A cepa de T. cubensis foi coletada na Floresta Nacional de Tef?? (FLONA) e determinada a atividade enzim??tica a partir da fermenta????o submersa e semi-s??lida. Como substrato testou-se serragem da madeira de Simarouba amara e baga??o-de-cana de a????car. Como indutores utilizou-se sulfato de cobre, ABTS e meio mineral. A lacase foi purificada por meio de cromotografia de troca i??nica e caracterizada quanto ?? massa (SDS-PAGE e zimograma) e determinados os par??metros cin??ticos Km e Vmax. A atividade de lacase mais expressiva ocorreu no meio com substrato de Simarouba amara (marup??) com 184 U/mL. O rendimento foi de 40% e fator de purifica????o de 1,5, com Vmax e Km de 33,33 ??mol.L-1 6,66 mM, respectivamente. A lacase proveniente de Trametes cubensis apresenta massa molecular aparente de, aproximadamente, 40 kDa. Nesse sentido, a partir do cultivo de Trametes cubensis ?? poss??vel obter lacases, contudo, estudos ainda s??o necess??rios no que se refere ao uso de diferentes indutores, visando uma potencializa????o dessa atividade.
Databáze: OpenAIRE
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