Repeated use of immobilized lipase for monoacylglycerol production by solid‐phase glycerolysis of olive oil

Abstract

By using immobilized lipase for production of monoacylglycerol (MAG) by solid-phase glycerolysis of fats and oils, the enzyme could be recovered easily from the reaction mixture and recycled to reduce the cost of the catalyst. Several support materials (CaCO 3 , CaSO 4 ·2H 2 O, Ca 2 P 2 O 7 , and Celite) were screened for immobilization of Pseudomonas sp. lipase by adsorption and tested for solid-phase glycerolysis of olive oil. Immobilization made the reuse of enzyme feasible. CaCO 3 proved to be the best support: 90% MAG (wt% in the glycerolfree reaction mixture after 72 h of reaction time) was obtained until the fifth use, 80% after the seventh use, and 60% after the tenth use. The same support was found suitable for immobilization of two other bacterial lipases from Chromobacterium viscosum and Pseudomonas pseudoalkali. JAOCS 74, 445-450 (1997).KEY WORDS: Bacterial lipases, CaCO 3 , CaSO 4 ·2H 2 O, Ca 2 P 2 O 7 , Celite, enzyme adsorption, glycerolysis, immobilized lipase, monoacylglycerol, reuse of immobilized enzyme.Monoacylglycerols (MAG) of saturated or unsaturated fatty acids are the most commonly used surfactants in the food industry. Their emulsifying properties are employed also in preparation of cosmetics and pharmaceutical products, manufacture of alkyd resins, detergents, etc. (1).They are produced industrially by transesterification of fats and oils with an excess of glycerol at high temperatures under alkaline catalysis. Depending on the glycerol excess and on the temperature, pressure and reaction time, equilibrium reaction mixtures of different compositions are obtained. The products contain 35-60% MAG, 35-50% diacylglycerols, 1-20% triacylglycerols,1-10% free fatty acids and their alkali metal salts, and 1-10% glycerol after ca. 1 h at 200-250°C (2). High-concentration MAG are obtained from the mixtures by molecular distillation. Use of the distilled product has increased with the desire to use purer and better products and because of their enhanced properties.The most serious drawbacks of the method are the low yields obtained and the high temperatures employed. The high temperatures lead to partial degradation of the products (formation of dark-colored by-products, burnt taste, peroxidation, and polymerization) and high energy consumption.Recently, lipase-catalyzed glycerolysis and direct esterification for the biosynthesis of partial glycerides are increasingly being studied as possible alternatives to the classical method (3-7). The main reasons are the higher yields achieved and much milder reaction conditions, resulting in products of higher quality and less energy costs. Furthermore, an enzyme with positional, substrate, or stereo selectivity can yield special products that can be used as intermediates for the syntheses of tailored triglycerides, enantiomeric acylglycerols, etc.Previous studies (8-12) have demonstrated that solidphase glycerolysis of fats and oils, catalyzed by lipases, may be an advantageous approach for obtaining MAG. In this process, the yield of MAG is greatly influ...

Enzymes as biocatalysts in the modification of natural lipids

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Use of Isopropanol as a Modifier in a Hexane‐Acetonitrile Based Mobile Phase for the Silver Ion HPLC Separation of Positional Isomers of Triacylglycerols Containing Long Chain Polyunsaturated Fatty Acids

“…Lipases used were as follows: Lipozyme (Rhizomucor miehei, 1,3-specific) was kindly donated by Novo Nordisk (Chiba, Japan); Liposam (Pseudomonas pseudoalkali, non-specific) was donated by Showa Denko (Tokyo, Japan); Lipase from Pseudomonas sp. KWI-56 was immobilized on CaCO 3 powder (PSL) as described by Rosu et al [21]. Single cell oil produced by a marine microbe, Shizochytrium sp.…”

Soy-Based Polyols and Polyurethanes

“…Lipases which have been known to catalyze soy-based polyol production such as Pseudomonas sp. and Candida rugosa have been tested in different immobilized support material systems such CaCO 3 , CaSO 4 ·2H 2 O, Ca 2 P 2 O 7 , Celite, polyurethane foam, Eupergit®C, bentotite, alginate beads, silica gel, porous glass surface, and many others (Kuncova et al, 2002;Yesiloglu, 2005;Knezevic et al, 2006;Rosu et al, 1997;Awang et al, 2007).…”