Produção de células íntegras de Penicillium citrinum para aplicação na hidrólise do óleo de soja

Abstract

The application of enzymatic technology in oils and fats has been consolidated as an attractive alternative for the substitution of traditional chemical processes.The hydrolysis of vegetable oils is a process by which several compounds of industrial interest can be obtained, allowing to add value both the production chain and the final products.This study aimed to the production of an active biocatalyst and stable from whole cells of the filamentous fungus Penicillium citrinum for application in the hydrolysis reaction of vegetable oils.The whole cells containing intracellular lipase attached to the mycelium were produced in free and immobilized form polyurethane foams cut into cubes 6 mm edge. The biocatalysts were characterized as their biochemical and kinetic properties including optimum values of pH, temperature and kinetic constants. The free cells showed optimal values of activity at pH 7.5 and temperature 45 ° C.O half-life obtained at 60 ° C was 1.81 h, with a maximum speed of 267,33 U / g. For immobilized cells, the best performance of intracellular lipase was at pH 8.0 and 40 ° C, with a maximum speed of 123,24 U / g and half-life at 60 ° C for 2,23 h.After the optimum conditions of cell performance were determined, the next stage of the work consisted in hydrolysis of soybean oil using the free cells, evaluating the effects of temperature, pH, agitation and solvent use.In order to improve system productivity, the temperature parameters and ratio by weight oil / buffer were optimized through experimental design, in search of better conditions for reaction. The optimized conditions predicted by the experimental design was 37 ° C and weight ratio oil / 20% buffer to give a maximum percentage of hydrolysis of 38.6% after 3 hours of reaction using 10% m / m biocatalyst in free form in the presence of emulsifying agent. The next step explored the use of ultrasonic waves in the hydrolysis of soybean oil with significant increases in the degree of maximum hydrolysis in a time period of 9 h.The values of 77 and 96% of hydrolysis degree with and without emulsifying agent were obtained, evidencing the positive influence of the ultrasonic waves in the reaction.The chromatographic analysis of the hydrolyzate showed that the P. citrinumcells provided the main fatty acids present in soybean oil, predominantly C18:1 and C18:2 fatty acids.The influence of vegetable oils containing different compositions in fatty acids (olive, canola and sunflower) was also evaluated in the hydrolysis reaction, being verified the preference of the free biocatalyst in the conversion to long chain fatty acids present in sunflower oil.At the end, the performance of the immobilized biocatalyst was investigated in the hydrolysis reaction of soybean oil and showed promising results, with a maximum hydrolysis degree of 55.7% in 12 hours of reaction.The results obtained in this work demonstrate that the use of intact cells as a biocatalyst in hydrolysis reactions of vegetable oils is a promising alternative to conventional catalysis, especially in the production of polyunsaturated fatty acids, such as omega 6.