Abstract
Fusarium verticillioides lipases were purified in a “cascade” method using octadecyl Sepabeads and octyl Sepharose resins, which led to the isolation of two proteins with lipolytic activities. Lip 1 was purified after octyl Sepharose adsorption presenting 30.3 kDa and, Lip 2 presented 68.0 kDa after octadecyl adsorption. These immobilization processes resulted in an increase of 3-fold in activity of each immobilized enzyme. These enzymes presented optima of pH of 5.0 and 6.0, respectively and temperature at 40 °C. They were thermostable at 40 °C and both remained more than 50% of its activity at the pH range of 5.0 to 7.0, with 180 min of incubation. The sardine oil hydrolysis showed higher EPA/DHA ratio. Concerning the ethanolysis reaction, Lip 2 showed higher conversion (5.5%) and both lipases showed activity in the release of the S enantiomers from 2-O-butyryl-2-phenylacetic acid (mandelic butyrate acid) and HPBE hydrolysis. Lip 2 also demonstrated capacity of transesterification. These applications made these enzymes attractive for industrial application.
Highlights
Lipases are very interesting enzymes due to their ability to catalyze a wide range of reactions, such as synthesis, transesterification and ester hydrolysis in aqueous or organic media, accompanied with a high regio- or enantioselectivity
This study aims a high selective purification of two lipases from
It was important to test the behavior of F. verticillioides crude extract on different supports with increasing levels of hydrophobicity
Summary
Lipases are very interesting enzymes due to their ability to catalyze a wide range of reactions, such as synthesis, transesterification and ester hydrolysis in aqueous or organic media, accompanied with a high regio- or enantioselectivity The interest in this class of enzymes essentially increased due to their industrial and biotechnological applications in oil and fat hydrolysis and because of their capacity to recognize different synthetic or natural substrates. Catalysts 2018, 8, 84 pharmaceutical industries as well as the use of fatty acid esters synthesis as cosmetic ingredients or surfactants, pesticides and agro-chemistry synthesis are interesting [1,2,3,4,5] Due to these broad uses, they are attractive in biotechnological application and there is an increasing search for new microorganisms able to produce these enzymes with different specificities and stabilities. Another advantage is the enzyme purification which minimizes or eliminates the contamination of other proteins, as shown in this work
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