General Introduction Enzyme Catalysis and its relevance to environment Biocatalysis in ionic liquid Lipases (triacylglycerol acylhydrolases, E.C.

3.1.1.3) are ubiquitous enzymes that catalyze the hydrolysis of fats and oils. Lipases catalyze the hydrolysis of triacylglycerols to glycerol and free fatty acids. In eukaryotes, lipases are involved in various stages of lipid metabolism including fat digestion, absorption, and lipoprotein metabolism. In plants, lipases are found in energy reserve tissues. Lipases are also produced by microorgaisms. Lipases have great importance in food,chemical,phermaceutical and several other industries.

Rview of Literature In 1856, Claude Bernard first discovered lipase in pancreatic juice as an enzyme that hydrolysed insoluble oil droplets and converted them to soluble products. Lipases have traditionally obtained from animal pancreas. Initial interest in microbial lipases was generated because of a shortage of pancreas and difficulties in collecting available material. The presence of lipases has been observed as early as in 1901 for Bacillus prodigiosus, B. pyocyaneus and B. fluorescens which represent todays best studied lipase producing bacteria now named Serratia marcescens, Pseudomonas aeruginosa and Pseudomonas fluorescens, respectively. Lipases differ greatly as regards both their origins (which can be bacterial, fungal, mammalian, etc.) and their properties and they can catalyze the hydrolysis, or synthesis, of a wide range of different carboxylic esters and liberate organic acids and glycerol. Lipases from a large number of bacterial, fungal and plant and animal sources.

Lipases catalyed reactions

The biological function of lipases to catalyse the hydrolysis of triglycerides to free fatty acids, diglycerides, monoglycerides and glycerols. Most of these substrates are insoluble in water so, reactions take place in emulsions. Lipases are well adapted for that and activated in the presence of an interface between an aqueous and organic phase (Brokeman 1984).

Lipase catalysed esterification and transesterification reactions These catalysed

based reactions in organic solvents provide new approches of chemists. Reversiblity of these reactions provides several difficulties regarding enzyme activity. To solve this problem, vinyl acetate was introduced as a acylating agent to being irreeversible by formation of acetaldehyde.

Other lipase based catalytic reactions -

Lipase Catalytic centre

The three dimension structure of lipases is determined by X-ray cristallography. All the lipases members come under /-hydrolase fold family. i.e., Structure is composed of a core of predominantly parallel strands surrounded by helices. The composition of catalytic centre of lipases with an /-hydrolase fold includes three amino acids (Serine, Histidine, Aspartate/Glutamate). Asp and Glu is negative charged residue which allow the His residue to acts as a general base which can remove a proton from the hydroxyl group of the active site Ser. Nucleophilic alxoxides ion generated ser residue is proposed to attack the carbonyl group of the esterified substrate forming an acyl enzyme intermediate. Oxyanion hole is another important component of catalytic machenisms. Which is composed of properly arranged H- bond donners ( mostly main chain NH groups). The oxyanion hole helps to stabilize a reaction intermediate during catalysis when the carbonyl oxygen carries a partial negative charge. The active serine residue of lipases is embedded in the short consensus sequence GXSXG (with X being any amino acid), a motif also found in esterases, thioesterases and proteases. The active site of lipases in the 'closed' form is shielded from the surface by protective surface loops called the 'lid'. Upon activation, the lid undergoes a conformational rearrangement exposing the active site serine and creating the active, open form of the enzyme. Both, the open and the closed form of lipases have been observed in X-ray structures of lipases.

Various type of lipases used

Lipases are widely used in the processing of fats and oils, detergents and degreasing formulations, food processing, the synthesis of fine chemicals and pharmaceuticals, paper manufacture, and production of cosmetics, and pharmaceuticals (Rubin and Dennis, 1997, Kazlauskas and Bornscheuer, 1998). Lipase can be used to accelerate the degradation of fatty waste (Masse et al., 2001) and polyurethane (Takamoto et al., 2001). Major applications of lipases are summarized in Table 2. Most of the industrial microbial lipases are derived from fungi and bacteria.