Characteristics of a Newly Isolated Lipase from Thermophilic Burkholderia Pseudomallei.docVIP

Characteristics of a Newly Isolated Lipase from Thermophilic Burkholderia Pseudomallei 　　Abstract. A lipase producing thermophilic Burkholderia pseudomallei bacterial strain was isolated from Saudi Arabian environment. Based on this strain, a lipase protein was purified to homogeneity 128.2 fold as a fusion protein with glutathione S-transferase. SDS- PAGE of the purified enzyme revealed it has Mr of 32 kDa. The recombinant lipase was efficiently immobilized in calcium alginate gelatin composites. The optimum temperature for free enzyme was recorded at 65 ℃. However the immobilized enzyme exhibits the highest activity at 70℃. 　　Key words: Cloning, Expression, Purification, Immobilization, Characterization, Glutathione S transferase. 　　1. Introduction 　　The production of fatty acids by the hydrolysis of natural oils and fats is a very important component in the economic utilization of these naturally produced renewable raw materials. These products include oils from corn, sunflower, palm, coconut, olives and rice bran. A significant number of high value products require fatty acids in their manufactures. These include coatings, adhesives, specially lubricating oils, shampoos and other personal care products. Oils and fats are part of a group of compounds known as fatty esters or triglycerides, and their hydrolysis essentially involves reactions with water to produce valuable free fatty acids and glycerol [1]. Enzymatic hydrolysis of triglycerides may be carried out at ambient conditions. Microorganisms produce different classes of lipolytic enzymes, including lipases (EC 3.1.1.3), which specifically catalyze the hydrolysis of ester linkages of long chain triglycerides at an oil-water interface [2]. Lipases are currently attracting enormous attention because they constitute the most important group of biocatalysts for biotechnological applications [3-5]. Most of the lipases used in industry are microbial enzymes, of both fungal and bacterial origin [6,7]. Bact