Cutinase production by fusarium oxysporum utilizing agro-industrial by products

Orientador: Gabriela Alves MacedoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Cutinases são enzimas amplamente encontradas na natureza, podendo ser obtidas de fontes vegetais ou microorganismos. Trata-se de uma esterase que atua tanto como esterase quanto como lipase, capaz de hidrolisar uma ampla variedade de ésteres sintéticos e triglicerídeos. Este perfil de atividade oferece-lhe um número de aplicações importantes nas indústrias como detergentes de roupas e louças, alimentos, fármacos, agroquímicos e cosméticos. Utilizando a fermentação em estado sólido (FES) é possível a produção de uma enzima com características interessantes de catálise, com baixo custo de produção e bom rendimento, e que possa ser empregada na indústria de modo geral. Este trabalho utilizou FES para produção de cutinase por Fusarium oxysporum utilizando sub-produtos agrícolas visando à seleção dos melhores meios e indutores para produção da enzima (utilizando fontes de carbono e minerais), otimização das condições de cultivo (avaliando os parâmetros temperatura e quantidade de água do meio além do estudo do inóculo), produção de cutinases utilizando diferentes substratos (farelo de trigo, casca de soja, farelo de arroz e produzida por meio líquido) e comparação entre eles, além da aplicação das enzimas em reações de esterificação para separação enantiomérica de substâncias racêmicas como o 2-octanol e o Ibuprofeno. De um modo geral o trabalho mostrou a produção de enzima de grande interesse comercial usando substratos diferentes e seu comportamento frente a reações de esterificação. Três meios sólidos foram selecionados (farelo de trigo, casca de soja e farelo de arroz) e suas condições de cultivo otimizadas (temperatura 28ºC e quantidade de água do meio 100%). Nos testes de aplicação das cutinases, para resolução enantiomérica do 2-octanol todas as cutinases tiveram bons valores de esterificação e enantioseletividade com destaque para farelo de trigo (76,4% de esterificação após 200 horas) e casca de soja (71,9% de esterificação após 200 horas). Para resolução do fármaco racêmico Ibuprofeno as cutinases mais efetivas foram as produzidas por farelo de arroz (5,1% de conversão após 200 horas) e meio líquido (2,9% de conversão após 200 horas). Esses resultados mostraram a diferença entre o comportamento de cutinases, do mesmo microorganismo, produzidas em diferentes meios, e a grande possibilidade de utilização de cutinases para processos de interesse industrialAbstract: Cutinases enzymes are often found in nature and may be obtained from plant sources or microorganisms. This is an enzyme that acts both as esterases and as lipases, capable of hydrolyzed a several variety of synthetic esters and triglycerides. This activity profile gives a number of important applications in industries such detergents, clothes and dishes, foods, drugs, chemicals and cosmetics. Using solid state fermentation SSF it is possible production of an enzyme interesting of catalysis with low production cost and good yields, and it can be employed in the industries. This work used SSF for the cutinase production by Fusarium oxysporum using agro-industrial sub-products to find selection of the best media and inducer to enzyme production (using sources of carbon and minerals), optimization of fermentation conditions (modifying the parameters temperature and water amount and after the inoculum study), cutinases production using different substrates (wheat bran, soy rind, rice bran and produced by liquid medium), comparison among them and enzymes application in esterification reactions to enantiomeric separation of racemic substances such 2-octanol and drug Ibuprofen. In general the work showed enzyme production of great commercial interest using various substrates and their behavior in esterification reactions. Three solid media were selected (wheat bran, soy rind, and rice bran) and their fermentation conditions optimized (temperature 28ºC and water amount 100%). In tests of cutinases application for enantiomeric resolution of 2-octanol all cutinases had good values to esterification and enantioselectivity and the best results was using wheat bran (76.4% of esterification after 200 hours) and soy rind (71.9% of esterification after 200 hours). The test using racemic drug Ibuprofen the more effective resolution was found with cutinase produced by rice bran (5.1% of esterification after 200 hours) and liquid medium (2.9% of esterification after 200 hours). These results showed the difference among behavior of cutinases using same strain, produced by different media, and the great possibility of cutinases with interesting catalysis characteristics and it can be employed in the industryMestradoMestre em Ciência de Alimento

Fungal cutinases: properties and industrial applications - review

Cutinases (EC 3.1.1.74) are also known as cutin hidrolases. These enzymes share catalytic properties of lipases and esterases, presenting a unique feature of being active regardless the presence of an oil-water interface, making them interesting as biocatalysts in several industrial processes involving hydrolysis, esterification and trans-esterification reactions. They are also active in different reaction media, allowing their applications in different areas such as food industry, cosmetics, fine chemicals, pesticide and insecticide degradation, treatment and laundry of fiber textiles and polymer chemistry. The present review describes the characteristics, potential applications and new perspectives for these enzymes.21182123Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Cutinases fúngicas: propriedades e aplicações industriais

Cutinases (EC 3.1.1.74) are also known as cutin hidrolases. These enzymes share catalytic properties of lipases and esterases, presenting a unique feature of being active regardless the presence of an oil-water interface, making them interesting as biocatalysts in several industrial processes involving hydrolysis, esterification and trans-esterification reactions. They are also active in different reaction media, allowing their applications in different areas such as food industry, cosmetics, fine chemicals, pesticide and insecticide degradation, treatment and laundry of fiber textiles and polymer chemistry. The present review describes the characteristics, potential applications and new perspectives for these enzymes

Lipase and esterase: to what extent can this classification be applied accurately?

Enzyme technology is an ever-growing field of knowledge and, in recent years, this technology has raised renewed interest, due to the search for new paradigms in several productive processes. Lipases, esterases and cutinases are enzymes used in a wide range of processes involving synthesis and hydrolysis reactions. The objective of this work was to investigate and compare the specific lipase and esterase activities of five enzymes - four already classified as lipases and one classified as cutinase - in the presence of natural and synthetic substrates. All tested enzymes presented both esterase and lipase specific activities. The highest specific esterase activity was observed for Aspergillus 1068 lipase in natural substrate and for F. oxysporum cutinase in synthetic substrate, while the highest specific lipase activity was observed for Geotrichum sp. lipase in natural substrate and for F. oxysporum cutinase in synthetic substrate. These results display some interface-independent lipolytic activity for all lipases tested. This is in accordance with the rationale that a new and broader definition of lipases may be necessary