Industrial Applications of Lipase

FIGURE 2.6 Reactions catalyzed by lipases (a) hydolysis, (b) esterification, (c) interesterification, (d) acidolysis, (e) alcoholysis, and (f) aminolysis. 

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The resolution of chiral alcohols through lipase-mediated enantioselective acylation (transesterification) is one of the major industrial applications of lipases [50]. Hence, the effects of ionic liquid reaction media on the resolution of various... 

In addition to hydrolyzing carboxylic ester bonds, lipases can catalyze a variety of esterification reactions in non-aqueous media. Industrial applications of lipases have focused on their enantioselectivity, regioselectively and substrate... 

Hasan F, Shah AA, Hameed A (2006) Industrial applications of lipases. Enzyme Microb Technol... 

Vulfson EN (1994) Industrial applications of lipases. In Woolley P, Peterson SB (eds). Lipases their structure, biochemistry and applications. Cambridge University Press, Cambridge, pp 271-288... 

Lipases are hydrolytic enzymes that have received extensive attention in food, pulp and paper and fuels industries. They can be found in microorganisms, plants, and animals with ability to breakdown of lipids. In this chapter, lipases are introduced and there is a brief discussion about their functions. This is followed by a short description of their main sources, structure, and features with an emphasis on their specificity and interfacial activity. The chapter focuses on microbial lipases, which are usually preferred for commercial applications due to their favorable properties, easy extraction, and unlimited supply. The chapter concludes with a discussion on the various industrial applications of lipases and properties improvement. 

Selectivity improvement is a critical requirement for industrial applications of lipase. This includes substrate selectivity, stereoselectivity, regioselectivity, and enantio-selectivity. Adsorption of Candida rugosa on celite was reported to enhance the stability of lipase and improve its enantioselectivity up to 3-fold (Ogino, 1970). Entrapment in cellulose acetate-Ti02 gel fiber improved the selectivity of Rhizomucor miehei lipase in the hydrolysis of 1,2-diacetoxypropane (Ikeda and Kurokawa, 2001). Also the enantioselectivity of pegylated P. cepacea lipase was increased 3-fold by... 

Schmid, R.D. and Verger, R., Lipases interfacial enzymes with attractive applications. Angew. Chem. Int. Ed., 1998, 37, 1608-1633 Hasan, F., Shah, A.A. and Hameed, A., Industrial applications of microbial lipases. Enzyme. Microb. TechnoL, 2006, 39, 235-251. 

Shimada, Y., Watanabe, Y., and Nagao, T. 2005a. Application of lipases to industrial-scale purification of oil- and fat-related compounds. In C. T. Hou (Ed.), Handbook of Industrial Biocatalysis (Chap. 8). Boca Raton, FL CRC Press, Taylor Francis Group. 

A new industrial application for lipases has been developed in the field of renewable energy. Lipases are used for methylation of plant oils such as rapeseed oil. The resulting methylated oils are used as a replacement or supplement for fossil fuel-derived diesel. As of today, the costs of the biocatalyst, however, prevent its commercial application and chemical processes such as saponification of fats with sodium methoxide are currently preferred. 

There is considerable worldwide interest in the application of lipases for fat modification and ester synthesis. In systems with low water activity, lipases are able to catalyze ester synthesis as well as interesterification reactions in fats. The lipase reactions under low water activity can be performed in several ways. We consider batch or continuous reactions in melted fats by use of immobilized lipases as the most relevant way for the industry. 

This chapter consequently focuses on the application of enzymes for the selective cleavage of esters, amides and nitriles [2], Out of all the reported industrial applications of enzymes these type of hydrolyses constitute more than 40% [3], Enzymatic hydrolyses are often performed because of the enantioselectivity of enzymes, and in particular of the lipases that are used for the production of enantiopure fine chemicals. 

Chemical interesterification is conveniently achieved by using alkali metal methylates as a catalyst. Microbial lipases are also used as biocatalysts in enzymatic interesteiification. In contrast to the chemical process, the enzymatic process can be more selective if an enzyme with positional specificity is used, but this reaction is usually much slower and more sensitive to reaction conditions. Recent developments in lipase-catalyzed interesterification have resulted in new industrial applications of this process (255). Nevertheless, the high costs of enzymes and process equipment may limit widespread adoption of this process. 

An application of lipases in the pharmaceutical industry can he found in the manufacture of Ihuprofen (Houde et al., 2004). Ihuprofen is a racemic mixture, where the (S)-ihuprofen molecules is 160 times more potent that the (R)-ihuprofen one. Resolution of racemic ihuprofen can he achieved hy esterification of (S)-ibuprofen with methanol or hutanol, in organic media using lipase, leading to synthesis of the corresponding (S)-ester. This ester can be separated from (R)-ibuprofen and chemical transformed to (S)-ibuprofen. 

The types of enzymes used by organic chemists vary widely and include such well-known biocataiysts as lipases, esterases, oxidoreductases, oxinitrilases, transferases and aldolases [4]. An example which illustrates the industrial application of a lipase concerns the kinetic resolution of a chiral epoxy ester used as the key intermediate in the synthesis of the calcium antagonist Diltiazem, a major therapeutic in the treatment of high blood pressure [6] (Fig. 1). In developing the industrial process for the production of this drug, many different lipases were screened, but only the bacterial lipase from Serratia marescens showed both a sufficiently high activity and enantioselectivity. The intermediate is produced industrially on a scale of 50 tons/year. 

Schoevaait R, Wolbers MW, Golubovic M et al. (2004) Preparation, optimization and structures of cross-Unked enzyme aggregates (CLEAs). Biotechnol Bioeng 87 754-762 Schulz T, Plesis J, Schmid RD (2000) Stereoselectivity of Pseudomonas cepacia lipase toward secondtuy alcohols a quantitative model. Protein Sci 9 1053-1062 Secundo F, Cturea, G, Tarabiono C et al. (2006) The lid is a stmctural and functional determinant of lipase activity and selectivity. J Mol Catal B Enzym 39 166-170 Seitz EW (1974) Industrial application of microbial lipases a review. JAOCS 51(2) 12-16 Sharma R, Chisti Y, Baneijee UC (2(X)1) Production, purification, characterization and application of lipases. Botechnol Adv 19 627-662... 

Hasan, R, Shah A. A., and Hameed A., Industrial applications of microbial lipases. Enzyme Microbiol. Technol., 39, 235, 2006. 

Lipases. The major application of lipases in the dairy industry is in the production of Italian cheeses, for example, Romano and Provo-lone. These cheese varieties have a characteristic piquant flavour due to short-chain fatty acids liberated from the milk fat by lipases. The lipases used are mainly from oral tissues because these have a higher specificity than those from microbial sources. A considerable amount of information (Shahani, 1975) has been accumulated on the characteristics of lipolytic systems. 

When the reaction yield was lower than 50%, a temperature drop from 60 to 40 °C diminished the E and enantiomeric excess, but a similar trend was not observed in the reactions at 23 and 40 °C. The enantioselectivity of the lipase in reactions performed at 60 °C was higher (for 1, E >125) than that at 40 °C (for 1, E -25). Usually an E value of 100 is considered a minimum satisfactory value for the industrial application of an enzymatic process. Thus, in the resolution of 1, a 20 temperature increment from 40 to 60 °C made an otherwise inadequate reaction process a very efficient one. This effect was also observed for 2, where the E value increased fi om 8 to 60 with an increase in temperature from 40 to 60 C. 

Hamid NSA, Zen HB, Tein OB, Halifah YM, Saari N, Bakar FA (2003) Screening and identification of extracellular lipase-producing thermophilic bacteria from a Malaysian hot spring. World J Microbiol Biotechnol 19 961-968 Hasan F, Shah AA, Hameed A (2006) Industrial applications of microbial lipases. Enzyme Microb Technol 39 235-251... 

Further Industrial applications of the synthetic potential of lipases have been reported In the area of foods. Thus, immobilized lipase was efficiently used In a solvent-free reaction system to produce cocoabutter from palm oil and stearic acid (40). This process Includes Intermittent removal... 

Applications of lipases include ester hydrolysis, regioselective acylation or deacylation, interesterification, and resolution of racemic mixtures. Such reactions can be employed in conjunction with a variety of substrate types, ranging from glycerol derivatives to organometallics [6]. Consequently, lipases can be utilized in a range of industries, including the manufacture of... 

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