PRODUÇÃO DE FITASES E PROTEASES FÚNGICAS, APLICAÇÃO EM RAÇÕES E VALORES DIGESTÍVEIS PELA TILÁPIA-DO-NILO

RESUMO

SCREENING OF FUNGI PHYTASES PRODUCED BY SOLID STATE FERMENTATION WITH DIFFERENT AGRICULTURAL BY-PRODUCTS AS SUBSTRATE This study was carried out for screening the phytase production using agroindustrial residues as substrate for solid state fermentation (SSF) and several fungal strains. High phytase production was observed for most of the microorganisms studied, as well as very different biochemical characteristics, including activities at specific pH values. The phytases showed very distinct behavior of optimum and stability pH, optimal values of temperature and stability. A. niger showed the higher values of enzyme activity. Lyophilized enzyme produced by A. niger reached 8,575 U/g and yield of 826,096.50 U/Kg of substrate when fermented in wheat bran. Therefore, the substrate as well as the microorganism strain can affect the biochemical character of the enzyme produced. The high phytase activity and very distinct characteristics established, plus the low cost of substrates, make these fungal phytases potential alternatives for biotechnological industry. Keywords Fungal phytase, agroindustrial residues, solid state fermentation, biochemical characteristics NOVEL INEXPENSIVE FUNGI PROTEASES PRODUCTION BY SOLID STATE FERMENTATION AND CHARACTERIZATION A comparative study was carried out for proteases production using agroindustrial residues as substrate for solid state fermentation (SSF) of several fungal strains. High protease production was observed for most of the microorganisms studied, as well as very different biochemical characteristics, including activities at specific temperatures and a wide range of pH values. The proteases showed very distinct behavior of optimum pH, optimal values of temperature and stability at 50 oC. A. oryzae showed stability at all pH values studied. P. roquefortii and A. flavipes presented optimum activity at temperatures of 50 and 90 oC, respectively. Lyophilized protease from A. oryzae reached 1,251.60 U/g and yield of 155,010.66 U/Kg of substrate. Therefore, the substrate as well as the microorganism strain can affect the biochemical character of the enzyme produced. The high protease activity and stability established plus the low cost of substrates, make these fungal proteases potential alternatives for the biotechnological industry. Keywords Fungal protease, agroindustrial residues, solid state fermentation, PRODUCTION OF Aspergillus niger 40018 ENZYMES WITH DIFFERENT AGROINDUSTRIAL CO-PRODUCTS AS SUBSTRATES Utilization of agricultural co-products as substrate for solid state fermentation (SSF) was studied, aimed the achievement of phytases and proteases from Aspergillus niger with different biological characteristics. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Protease produced with wheat bran and soybean bran was stable in the range of pH 7.0 - 9.0, however with higher production yield in wheat bran. As a result, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Keywords phytase, protease, Aspergillus niger, isozymes, Solid fermentention. DIGESTIBLE VALUES FOR NILE TILAPIA (Oreochromis niloticus) USING FUNGAL PHYTASES AND PROTEASES PRODUCED BY SOLID STATE FERMENTATION The use of enzymes in animal feed is common practice nowadays and the use of exogenous microbial enzymes has shown good results for improving the efficiency of nutrients utilization. The ingredients used for fish feed formulation aim to provide nutrients to meet their requirements, and affect the availability in the animal intestinal tract. In this sense, the goal of this work was to add low cost fungal phytases and proteases produced by Solid State Fermentation using agroindustrial co-products to improve nutritional availability of plant based diets for Nile tilapia. Concentrated enzyme extract of phytase, protease and both enzymes was added to diets with soybean meal or corn gluten as test ingredients for further analysis of nutrients digestibility. Phytase supplementation increased dry matter, protein, energy, lipids and phosphorous utilization for soybean meal (P¿0.05). Protease affects energy, lipids, phosphorous and magnesium availability for soybean meal (P¿0.05). As for corn gluten, the enzymes did not show improvement for nutrients and minerals utilization. When the enzymes were combined in the diets they did not show improved as they were used alone. These low cost enzymes are an alternative as additives to improve soybean meal digestibility in fish plant based diets. Keywords Phytase, Protease, Solid State Fermentation (SSF), Oreachromis niloticus, Nutritional requirements,

ABSTRACT

SCREENING OF FUNGI PHYTASES PRODUCED BY SOLID STATE FERMENTATION WITH DIFFERENT AGRICULTURAL BY-PRODUCTS AS SUBSTRATE This study was carried out for screening the phytase production using agroindustrial residues as substrate for solid state fermentation (SSF) and several fungal strains. High phytase production was observed for most of the microorganisms studied, as well as very different biochemical characteristics, including activities at specific pH values. The phytases showed very distinct behavior of optimum and stability pH, optimal values of temperature and stability. A. niger showed the higher values of enzyme activity. Lyophilized enzyme produced by A. niger reached 8,575 U/g and yield of 826,096.50 U/Kg of substrate when fermented in wheat bran. Therefore, the substrate as well as the microorganism strain can affect the biochemical character of the enzyme produced. The high phytase activity and very distinct characteristics established, plus the low cost of substrates, make these fungal phytases potential alternatives for biotechnological industry. Keywords Fungal phytase, agroindustrial residues, solid state fermentation, biochemical characteristics NOVEL INEXPENSIVE FUNGI PROTEASES PRODUCTION BY SOLID STATE FERMENTATION AND CHARACTERIZATION A comparative study was carried out for proteases production using agroindustrial residues as substrate for solid state fermentation (SSF) of several fungal strains. High protease production was observed for most of the microorganisms studied, as well as very different biochemical characteristics, including activities at specific temperatures and a wide range of pH values. The proteases showed very distinct behavior of optimum pH, optimal values of temperature and stability at 50 oC. A. oryzae showed stability at all pH values studied. P. roquefortii and A. flavipes presented optimum activity at temperatures of 50 and 90 oC, respectively. Lyophilized protease from A. oryzae reached 1,251.60 U/g and yield of 155,010.66 U/Kg of substrate. Therefore, the substrate as well as the microorganism strain can affect the biochemical character of the enzyme produced. The high protease activity and stability established plus the low cost of substrates, make these fungal proteases potential alternatives for the biotechnological industry. Keywords Fungal protease, agroindustrial residues, solid state fermentation, PRODUCTION OF Aspergillus niger 40018 ENZYMES WITH DIFFERENT AGROINDUSTRIAL CO-PRODUCTS AS SUBSTRATES Utilization of agricultural co-products as substrate for solid state fermentation (SSF) was studied, aimed the achievement of phytases and proteases from Aspergillus niger with different biological characteristics. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Protease produced with wheat bran and soybean bran was stable in the range of pH 7.0 - 9.0, however with higher production yield in wheat bran. As a result, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Keywords phytase, protease, Aspergillus niger, isozymes, Solid fermentention. DIGESTIBLE VALUES FOR NILE TILAPIA (Oreochromis niloticus) USING FUNGAL PHYTASES AND PROTEASES PRODUCED BY SOLID STATE FERMENTATION The use of enzymes in animal feed is common practice nowadays and the use of exogenous microbial enzymes has shown good results for improving the efficiency of nutrients utilization. The ingredients used for fish feed formulation aim to provide nutrients to meet their requirements, and affect the availability in the animal intestinal tract. In this sense, the goal of this work was to add low cost fungal phytases and proteases produced by Solid State Fermentation using agroindustrial co-products to improve nutritional availability of plant based diets for Nile tilapia. Concentrated enzyme extract of phytase, protease and both enzymes was added to diets with soybean meal or corn gluten as test ingredients for further analysis of nutrients digestibility. Phytase supplementation increased dry matter, protein, energy, lipids and phosphorous utilization for soybean meal (P¿0.05). Protease affects energy, lipids, phosphorous and magnesium availability for soybean meal (P¿0.05). As for corn gluten, the enzymes did not show improvement for nutrients and minerals utilization. When the enzymes were combined in the diets they did not show improved as they were used alone. These low cost enzymes are an alternative as additives to improve soybean meal digestibility in fish plant based diets. Keywords Phytase, Protease, Solid State Fermentation (SSF), Oreachromis niloticus, Nutritional requirements,