RESUMO
Lipases (triacylglycerol hydrolases, EC 3.1.1.3) are a class of enzymes with high industrial importance. An option for the production of this enzyme is through fungal growth via solid-state fermentation (SSF). Thus, this research presents a study of lipase production by Penicillium roqueforti ATCC 10110 through SSF using cocoa bran residues (Theobroma cacao) as a substrate. To achieve maximum lipase production, fermentation time (0 to 120 h) and palm oil (PO) percentage (0 to 50%) were optimized through analysis of one factor at a time (OFAT), with lipase activity as the response. The amount of cocoa was fixed (5 g), the incubation temperature was maintained at 27 °C, and the moisture content was established at 70%. For a 72 h incubation, the highest enzyme activity achieved using SSF without adding PO was 14.67 ± 1.47 U g-1, whereas with PO (30%), it was 33.33 ± 3.33 U g-1, thus demonstrating a 44% increase in enzyme activity. Through the OFAT methodology, it was possible to confirm that supplementation with palm residue was efficient and maximized the lipase of P. roqueforti ATCC 10110.
Assuntos
Arecaceae/metabolismo , Cacau/metabolismo , Fermentação , Lipase/biossíntese , Penicillium/metabolismoRESUMO
Nitrilases and nitrile hydratases/amidases hydrolyze nitriles into carboxylic acids and/or amides, which are used in industrial chemical processes. In the present study, 26 microorganisms, including yeasts and filamentous fungi, in a minimum solid mineral medium supplemented with glucose and phenylacetonitrile were screened to evaluate their biocatalytic potential. Of these microorganisms, five fungi of the genus Aspergillus were selected and subjected to colorimetry studies to evaluate the production and distinction of nitrilase and nitrile hydratase/amidase enzymes. Aspergillus parasiticus Speare 7967 and A. niger Tiegh. 8285 produced nitrilases and nitrile hydratase, respectively. Nitrilase optimization was performed using a Box-Behnken design (BBD) and fungus A. parasiticus Speare 7967 with phenylacetonitrile volume (µl), pH, and carbohydrate source (starch:glucose; g/g) as independent variables and nitrilase activity (U ml-1 ) as dependent variable. Maximum activity (2.97 × 10-3 U ml-1 ) was obtained at pH 5.5, 80 µl of phenylacetonitrile, and 15 g of glucose. A. parasiticus Speare 7967 showed promise in the biotransformation of nitriles to carboxylic acids.
Assuntos
Aminoidrolases , Ensaios de Triagem em Larga Escala , Fungos , Nitrilas/metabolismo , Ácidos Carboxílicos/metabolismo , Aspergillus/metabolismo , GlucoseRESUMO
Solid state fermentation (SSF) simulates the natural conditions fungal growth, where the amount of water in the reaction medium must be restricted, thus limiting the use of liquid substrate. An analytical strategy to deal with this limitation is the design of blending with constraints. Thus, the objective of the work was to optimize two constrained waste mixtures for the production of lipase by Penicillium roqueforti ATCC 10110 under SSF, using different substrates that combine solid and liquid waste. For this, the best fermentation time was determined through a fermentative profile, afterwards a restricted-mix design with lower and upper limits of the components of mixture I (cocoa residue, solid palm oil residue and liquid palm oil residue) and II (cocoa residue, mango residue and palm oil residue liquid palm) was applied. By means of Pareto and contour graphs, the maximum production points of lipase in mixtures I (6.67 ± 0.34 U g-1) and II (6.87 ± 0.35 U g-1) were obtained. The restricted mixture design proved to be a promising tool in the production of lipase by P. roqueforti ATCC 10110 under SSF since the use of restrictions is useful when intending to combine solid and liquid residues in fermentation processes.
Assuntos
Resíduos Industriais , Lipase , Fermentação , Lipase/metabolismo , Óleo de Palmeira , PenicilliumRESUMO
OBJECTIVES: To immobilize Candida rugosa lipase in Accurel MP 1000 (CRL-AMP) by physical adsorption in organic medium and apply in the synthesis of wax esters dodecanoyl octadecanoate 1 and hexadecanoyl octadecanoate 2 in a heptane medium, as well as evaluating the stability and recyclability of CRL-AMP in six reaction cycles. RESULTS: The specific activity (Asp) for CRL-AMP was 200 ± 20 U mg-1. Its catalytic activity was 1300 ± 100 U g-1. CRL-AMP was used in the synthesis of esters in heptane medium with a 1:1 acid:alcohol molar ratio at 45 °C and 200 rpm. In synthesis 1, conversion was 62.5 ± 3.9% in 30 min at 10% m v-1 and 56.9 ± 2.8% in 54 min at 5% m v-1; while in synthesis 2, conversion was 79.0 ± 3.9% in 24 min at 10% m v-1, and 46.0 ± 2.4% in 54 min at 5% m v-1. Reuse tests after six consecutive cycles of reaction showed that the biocatalyst retained approximately 50% of its original activity for both reaction systems. CONCLUSIONS: CRL-AMP showed a high potential in the production of wax esters, since it started from low enzymatic load and high specific activities and conversions were obtained, in addition to allowing an increase in stability and recyclability of the prepared biocatalyst.