Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso

Correia, Thaylanna Cavalcante

Dissertação

Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso

Poultry farming is one of the sectors of great impact on the Brazilian economy. In recent years, there has been an increase in the production of broiler chicken, making this sector of the industry responsible for the generation of tons of feathers, which have polluting potential when improperly disp...

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Autor principal:	Correia, Thaylanna Cavalcante
Grau:	Dissertação
Idioma:	por
Publicado em:	Universidade Federal de Roraima 2023
Assuntos:	Biotecnologia Avicultura Resíduos Aspergillus sp. Enzimas microbianas Biotechnology Poultry farming Waste Microbial enzymes CNPQ::CIENCIAS BIOLOGICAS
Acesso em linha:	http://repositorio.ufrr.br:8080/jspui/handle/prefix/842

id	oai:repositorio.ufrr.br:prefix-842
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spelling	oai:repositorio.ufrr.br:prefix-8422023-06-01T14:05:32Z Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso Use of poultry residius in the production of proteases by filamentous fungus Correia, Thaylanna Cavalcante Vital, Marcos José Salgado http://lattes.cnpq.br/7855596789769104 Corrêa, Ana Paula Folmer http://lattes.cnpq.br/6615922848800872 Biotecnologia Avicultura Resíduos Aspergillus sp. Enzimas microbianas Biotechnology Poultry farming Waste Microbial enzymes CNPQ::CIENCIAS BIOLOGICAS Poultry farming is one of the sectors of great impact on the Brazilian economy. In recent years, there has been an increase in the production of broiler chicken, making this sector of the industry responsible for the generation of tons of feathers, which have polluting potential when improperly disposed of in the environment. The disposal of feathers can become an economic problem for companies, which must invest in their correct destination to avoid possible environmental impacts. Given this, there is a need for sustainable alternatives for recycling these protein rich waste. The objective of this study was to investigate the production of proteases secreted by filamentous fungus, isolated from the soil of the Amazon Forest in Roraima, using chicken feathers as the only source of energy, to characterize the strain as to the optimum temperature and pH of production, as well as the effects chemical products, in addition to evaluating the effect of different growth substrates on proteolytic activity. Initially, a screening of 40 strains of filamentous fungi obtained from soil samples from Parna Viruá (preserved in the Culture Collection of the Microbiology Laboratory-PRONAT) was carried out, in order to select an isolate with the potential for the production of extracellular protease. The identification of the selected fungus was carried out through macroscopic and microscopic observations, inoculating the isolate in specific culture media, which was identified as Aspergillus sp. The fungus was inoculated on milk agar and feather meal agar to verify the qualitative production of proteases and keratinases, respectively. The formation of halo indicated positivity for both enzyme productions. As for the quantitative assessment of proteolytic and keratinolytic activities, obtained through spectrophotometry at 420 nm, the results indicated the ability of Aspergillus sp. in producing both proteases and keratinases. The ability to secrete proteases was evaluated on different growth substrates (whole feather, feather meal, peptone, casein, chicken beak, hair and gelatin). The highest production of enzymes occurred in culture media with peptone and feather meal, indicating that Aspergillus sp. is a keratinolytic microorganism, capable of efficiently degrading keratinous materials. The protease showed optimal activity at pH 5.0 and temperature of 37 ° C. The enzymatic activity was enhanced with the addition of CaCl2, MnSO4, KCl, MgSO4 and CuSO4. The detergents Tween 20 and Triton x-100 tended to stimulate activity. The enzymes were resistant to organic solvents (methanol, acetone, butanol, acetonitrile, isoprapanol and DMSO), keeping the enzymatic activity close to the control (100%). The β-mercaptoethanol inhibitors and ethylenediaminetetraacetium acid (EDTA) did not inhibit proteolytic activity in enzymatic assays, suggesting that the enzymes present in the enzyme hydrolyzate are neither metalloproteases nor cysteine proteases. From the perspective of industrial microbiology, the results of this research suggest that the crude protease extract can potentially be used in the bioconversion of keratinous residues, considered difficult to break down in the environment, and the possibility of applying protein hydrolyzate in animal feed supplementation in use as biofertilizers. A avicultura é um dos setores de grande impacto na economia brasileira. Nos últimos anos, tem sido observado um aumento na produção de frangos de corte, fazendo com que este setor da indústria seja responsável pela geração de toneladas de penas, que possuem potencial poluente quando descartadas de forma inadequada no meio ambiente. O descarte das penas pode se tornar um problema econômico para as empresas, que devem investir na sua correta destinação para evitar possíveis impactos ambientais. Diante disto, surge a necessidade por alternativas sustentáveis para a reciclagem desses resíduos ricos em proteína. O objetivo deste estudo foi investigar a produção de proteases secretadas por fungo filamentoso, isolado de solo de Floresta Amazônica em Roraima, utilizando penas de frango como única fonte de energia, caracterizar a cepa quanto à temperatura e pH ótimos de produção, bem como os efeitos de produtos químicos, além de avaliar o efeito de diferentes substratos de crescimento sobre a atividade proteolítica. Inicialmente, foi realizado um screening de 40 linhagens de fungos filamentosos obtidos de amostras de solo do Parna Viruá (preservados na Coleção de Culturas do Laboratório de Microbiologia-PRONAT), a fim de selecionar um isolado com potencial para a produção de protease extracelular. A identificação do fungo selecionado, foi realizada por meio de observações macroscópicas e microscópicas inoculando o isolado em meios de cultura específicos, o qual foi identificado como Aspergillus sp. O fungo foi inoculado em meio ágar leite e ágar farinha de penas para verificar a produção qualitativa de proteases e queratinases, respectivamente. A formação de halo indicou positividade para ambas as produções enzimáticas. Quanto a avaliação quantitativa das atividades proteolítica e queratinolítica, obtidas através da espectrofotometria a 420 nm, os resultados indicaram a habilidade de Aspergillus sp. em produzir tanto proteases quanto queratinases. A capacidade de secretar proteases foi avaliada em diferentes substratos de crescimento (pena inteira, farinha de penas, peptona, caseína, bico de frango, cabelo e gelatina). A maior produção de enzimas ocorreu em meios de cultivo com peptona e farinha de penas, indicando que Aspergillus sp. é um microrganismo queratinolítico, capaz de degradar materiais queratinosos com eficiência. A protease apresentou atividade ótima em pH 5,0 e temperatura moderada de 37 °C. A atividade enzimática foi potencializada com a adição de CaCl2, MnSO4, KCl, MgSO4 e CuSO4. Os detergentes Tween 20 e Triton x-100 tenderam a estimular a atividade. As enzimas foram resistentes aos solventes orgânicos (metanol, acetona, butanol, acetonitrila, isoprapanol e DMSO), mantendo a atividade enzimática próxima ao controle (100 %). Os inibidores β-mercaptoetanol e o ácido etilenodiaminotetracécio (EDTA) não inibiram a atividade proteolítica em ensaios enzimáticos, sugerindo que as enzimas presentes no hidrolisado enzimático não são metaloproteases nem cisteína proteases. Na perspectiva da microbiologia industrial, os resultados desta pesquisa sugerem, que o extrato da protease bruta pode ser potencialmente utilizado na bioconversão de resíduos queratinosos, considerados de difícil degradação no meio ambiente, e a possibilidade de aplicação do hidrolisado proteico na suplementação de ração animal e no uso como biofertilizantes. Agência 1 2023-02-27T19:09:33Z 2023 2023-02-27T19:09:33Z 2021 Dissertação CORREIA. Thaylanna Cavalcante. Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso. 2021. 65 f. Dissertação (Mestrado em Ciências Ambientais) - Programa de Pós-Graduação em Recursos Naturais, Universidade Federal de Roraima, Boa Vista, 2021. http://repositorio.ufrr.br:8080/jspui/handle/prefix/842 por Acesso Aberto Attribution-NonCommercial-ShareAlike 3.0 Brazil http://creativecommons.org/licenses/by-nc-sa/3.0/br/ Universidade Federal de Roraima Brasil PRPPG - Pró-reitoria de Pesquisa e Pós-Graduação PRONAT - Programa de Pós-Graduação em Recursos Naturais UFRR
institution	Repositório Institucional - Universidade Federa de Roraima - UFRR
collection	DSpace
language	por
topic	Biotecnologia Avicultura Resíduos Aspergillus sp. Enzimas microbianas Biotechnology Poultry farming Waste Microbial enzymes CNPQ::CIENCIAS BIOLOGICAS
spellingShingle	Biotecnologia Avicultura Resíduos Aspergillus sp. Enzimas microbianas Biotechnology Poultry farming Waste Microbial enzymes CNPQ::CIENCIAS BIOLOGICAS Correia, Thaylanna Cavalcante Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
topic_facet	Biotecnologia Avicultura Resíduos Aspergillus sp. Enzimas microbianas Biotechnology Poultry farming Waste Microbial enzymes CNPQ::CIENCIAS BIOLOGICAS
description	Poultry farming is one of the sectors of great impact on the Brazilian economy. In recent years, there has been an increase in the production of broiler chicken, making this sector of the industry responsible for the generation of tons of feathers, which have polluting potential when improperly disposed of in the environment. The disposal of feathers can become an economic problem for companies, which must invest in their correct destination to avoid possible environmental impacts. Given this, there is a need for sustainable alternatives for recycling these protein rich waste. The objective of this study was to investigate the production of proteases secreted by filamentous fungus, isolated from the soil of the Amazon Forest in Roraima, using chicken feathers as the only source of energy, to characterize the strain as to the optimum temperature and pH of production, as well as the effects chemical products, in addition to evaluating the effect of different growth substrates on proteolytic activity. Initially, a screening of 40 strains of filamentous fungi obtained from soil samples from Parna Viruá (preserved in the Culture Collection of the Microbiology Laboratory-PRONAT) was carried out, in order to select an isolate with the potential for the production of extracellular protease. The identification of the selected fungus was carried out through macroscopic and microscopic observations, inoculating the isolate in specific culture media, which was identified as Aspergillus sp. The fungus was inoculated on milk agar and feather meal agar to verify the qualitative production of proteases and keratinases, respectively. The formation of halo indicated positivity for both enzyme productions. As for the quantitative assessment of proteolytic and keratinolytic activities, obtained through spectrophotometry at 420 nm, the results indicated the ability of Aspergillus sp. in producing both proteases and keratinases. The ability to secrete proteases was evaluated on different growth substrates (whole feather, feather meal, peptone, casein, chicken beak, hair and gelatin). The highest production of enzymes occurred in culture media with peptone and feather meal, indicating that Aspergillus sp. is a keratinolytic microorganism, capable of efficiently degrading keratinous materials. The protease showed optimal activity at pH 5.0 and temperature of 37 ° C. The enzymatic activity was enhanced with the addition of CaCl2, MnSO4, KCl, MgSO4 and CuSO4. The detergents Tween 20 and Triton x-100 tended to stimulate activity. The enzymes were resistant to organic solvents (methanol, acetone, butanol, acetonitrile, isoprapanol and DMSO), keeping the enzymatic activity close to the control (100%). The β-mercaptoethanol inhibitors and ethylenediaminetetraacetium acid (EDTA) did not inhibit proteolytic activity in enzymatic assays, suggesting that the enzymes present in the enzyme hydrolyzate are neither metalloproteases nor cysteine proteases. From the perspective of industrial microbiology, the results of this research suggest that the crude protease extract can potentially be used in the bioconversion of keratinous residues, considered difficult to break down in the environment, and the possibility of applying protein hydrolyzate in animal feed supplementation in use as biofertilizers.
author_additional	Vital, Marcos José Salgado
author_additionalStr	Vital, Marcos José Salgado
format	Dissertação
author	Correia, Thaylanna Cavalcante
title	Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
title_short	Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
title_full	Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
title_fullStr	Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
title_full_unstemmed	Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
title_sort	utilização de resíduos da avicultura na produção de proteases por fungo filamentoso
publisher	Universidade Federal de Roraima
publishDate	2023
url	http://repositorio.ufrr.br:8080/jspui/handle/prefix/842
_version_	1787156677621972992
score	11.755432

Utilização de resíduos da avicultura na produção de proteases por fungo filamentoso

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