Immobilization of Lipases Produced by the Endophytic Fungus Cercospora kikuchii on Chitosan Microparticles

Abstract

This work studied the immobilization of Cercospora kikuchii lipases on chitosan microparticles by chemical attachment on chitosan acetate microparticles activated by glutaraldehyde (CAM) added before or after the enzyme and physical adsorption on highly deacetylated chitosan hydrochloride microparticles (CHM). Lipases covalently immobilized on pre-activated CAM showed better performance retaining 88.4% of the enzymatic activity, with 68.2% of immobilization efficiency (IE). The immobilized enzyme retained an activity of about 53.5 % after five reuses, using p-NPP as substrate. Physical adsorption of lipase onto highly deacetylated CHM showed 46.2 % of enzymatic activity and 28.6% of IE. This immobilized derivative did not lose activity up to 80 days of storage at 4°C, while lipases immobilized on pre-activated CAM maintained its activity up to 180 days at same conditions. Taken together the results indicate that chitosan microparticles provide an optimal microenvironment for the immobilized enzyme to maintain good activity and stability.

Phytopathogenic Cercosporoid Fungi—From Taxonomy to Modern Biochemistry and Molecular Biology

“…Nonetheless, partial purification and biochemical characterization of lipase from C. kikuchii has been performed including enzyme stabilization studies with the spray drying technique [ 123 , 124 ]. The enzyme was also subjected to different immobilization strategies using chitosan microparticles, and appeared to be suitable for industrial applications [ 125 ]. Moreover, laccase from Cercospora SPF-6 has been identified recently and used for dye decolorization after immobilization procedure [ 126 ].…”

Enzymatic Transesterification of Coconut Oil Using Chitosan-Immobilized Lipase Produced by Fluidized-Bed System

“…Therefore, biocatalyst immobilization plays a crucial role within applied enzymology, especially for biodiesel production, and the main reason is the ability to isolate the enzyme–support system from the reactive medium and to reuse it, increasing the rate yields and consequently reducing the production cost, easier downstream operations (easy product separation and simple glycerol recovery), the potential to run continuous processes via packed-bed reactors, and improvement of activity and stability in terms of mechanical, chemical, and thermal properties of the lipase. , The enzymes’ properties would be affected depending on the type of immobilization technique and support used. Several methods for enzyme immobilization have been used by many researchers including adsorption onto solid supports, covalent attachment, and entrapment within sol–gel compounds. − The choice of the support/carrier is a relevant aspect influencing the immobilized biocatalyst properties and consequently the feasibility and process cost . An overview of the recent scientific literature reveals that a wide range of materials have been applied for enzymes immobilization and the high cost of commercial matrices (synthetic polymers and silica-based supports) has led several researchers to look for cheaper substitutes such as sugar cane bagasse, CaCO 3 , rice husk, , chitin, and chitosan .…”

“…Several methods for enzyme immobilization have been used by many researchers including adsorption onto solid supports, covalent attachment, and entrapment within sol–gel compounds. − The choice of the support/carrier is a relevant aspect influencing the immobilized biocatalyst properties and consequently the feasibility and process cost . An overview of the recent scientific literature reveals that a wide range of materials have been applied for enzymes immobilization and the high cost of commercial matrices (synthetic polymers and silica-based supports) has led several researchers to look for cheaper substitutes such as sugar cane bagasse, CaCO 3 , rice husk, , chitin, and chitosan . From these alternatives, the chitosan (a chitin derivative) appears to be more interesting since chitin is one of the most plentiful biopolymers on the planet.…”

“…11 An overview of the recent scientific literature reveals that a wide range of materials have been applied for enzymes immobilization and the high cost of commercial matrices (synthetic polymers and silica-based supports) has led several researchers to look for cheaper substitutes such as sugar cane bagasse, 11 CaCO 3 , 12 rice husk, 13,14 chitin, 15 and chitosan. 8 From these alternatives, the chitosan (a chitin derivative) appears to be more interesting since chitin is one of the most plentiful biopolymers on the planet. This material shows many benefits/advantages as enzyme immobilizing support: abundant and relatively inexpensive; several possibilities of chemical modification; high affinity toward the enzymes; biodegradability; and nontoxicity.…”