Abstract
Lipases are biocatalysts that may have distinct biochemical characteristics depending on the source. The combination of lipases from different sources with complementary characteristics is a viable strategy for increasing the enzymatic activity. In this study, fungal (Aspergillus niger 01 - CBMAI 2084) and plant (orange frit – orange peel fragment) lipases were analyzed separately and together in different concentrations. In addition, we evaluated the influence of organic solvents and ultrasonic effect on lipase activity, as well as substrate specificity [p-nitrophenyl butyrate (pNPB), p-nitrophenyl-laurate (pNPL) and p-nitrophenyl-palmitate (p-NPP)] and enzymatic immobilization in different supports (celite, silica, aluminum oxide, Lewatti, calcium alginate and gelatin). Increased enzyme activity was observed in formulations with higher concentration of fungal extract. The combination of 50% fungal extract and 25% plant extract increased about 55% lipase activity, showed the highest absolute lipase activity among all combinations and was selected for the following study. Plant extract showed the highest lipase activity in the hydrolysis of p-NPB and p-NPL, however, fungal extract showed the highest hydrolytic activity on p-NPP. When applied to synthetic substrates, the combination of plant and fungal extracts showed less stability and activity in synthetic substrates than isolated extracts, however lipase activity increased after 20s on ultrasound bath. Regarding to immobilization techniques, the adsorption on silica was the most efficient for all lipase extracts.
Highlights
Lipases are required for different applications and need to gather adequate biochemical characteristics and be obtained by more ecological processes (Abdallah et al 2021)
The fungal extract was obtained from the strain Aspergillus niger 01 (CBMAI 2084), this microorganism specie was registered under n.A3B348F in the platform of the National Genetic Heritage Management System (SISGEN), as recommended by the Brazilian Biodiversity Law (n. 13.123/15), in the laboratory it is preserved in PDA (Potato Dextrose Agar, KASVI®, Italy) slants
Lipases produced by Aspergillus niger 01 (CBMAI 2084) and orange frit can hydrolyze fatty acid of different chain lengths
Summary
Lipases are required for different applications and need to gather adequate biochemical characteristics and be obtained by more ecological processes (Abdallah et al 2021). Since lipases can be obtained from different sources, they can have totally different characteristics, differing in the type of catalyzed reaction, affinity for the substrate, Km, Vmax, temperature, pH, organic solvent tolerance, and ultrasonic influence (Bharathi & Rajalakshmi 2019, Khan et al 2015, Patil et al 2011, Seth et al 2014). Microbial lipases such as those extracted from Candida ssp.
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