Optimization of Lipase Activity Obtained from Dairy Industry Wastes

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Optimization of Lipase Activity Obtained from Dairy Industry Wastes

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Jaganmai. G & Rajeswari Jinka*

Department of Biochemistry, University College of Sciences, AcharyaNagarjuna University, Guntur, Andhra Pradesh, India

Abstract:- Optimisation of media components is very essential for the production of lipase to enhance its activity in order to meet wide variety of industrial applications. The enzyme production by the microorganisms in the environment is very low, hence it is essential to improve under required optimum growth conditions. This study mainly focuses on the enhancement of lipase production and its activity which includes the environmental, nutritional and biochemical factors of the media. In order to accomplish the enhanced activity, media parameters such as Lipid, and Carbon and Nitrogen sources along with other process parameters were altered. The lipase production mainly depends on nitrogen source, carbon source, metal ions, pH, and temperature and incubation time interval for media enrichment. The observations includes the screening of lipolytic bacteria by quantitative estimation of lipase by titrimetric method for isolated bacteria from dairy industry waste, confirmed on tributyrin agar and found to be highest lipase enzyme producer at an optimum pH 8, temperature 370C with sunflower oil as substrate where as the nitrogen and carbon sources found to be peptone and glucose respectively.

Key words: Bacteria, Tributyrin agar, Lipolysis, enrichment

I. INTRODUCTION:

Dairy industries around the world, produces wide range of products like cheese, milk, buttermilk etc. During the manufacturing of these products, the industries are involved in the production of solid and liquid wastes. The effluents which were collected from the diary industry consist of wide range of proteins, fats, and possibly other active substances. Micro biota are adapted to the waste water physiochemical conditions and produces wide range of enzymes like lipases, proteases etc. Some new active bacterial strains may be identified in waste water effluents from dairy industries.

Lipases are ubiquitous enzymes which are found in animals, plants, fungi, bacteria. Enzymes from microbial source are more stable than their corresponding plant and animal enzymes and the production is more convenient and safer. Microbial lipases have been produced by yeasts, fungi, and bacteria as extracellular, intracellular, and cell-bound enzyme. It has been estimated that only about 2% of the worlds microorganisms have been tested as source of enzymes (sahu etal. 2011).These enzymes are generally involved in increasing free fatty acid [FFA] concentration through the hydrolysis of fats or lipids. They are mainly involved in hydrolysis, which catalyses the breakdown of triacylglycerol to glycerol and fatty acids. Hence these enzymes are used to catalyze interesterification of fats and oils to modify glycerides. Lipase production is also used as a marker for pathogenicity in some medically important bacteria. Many varieties of lipases are produced from both gram positive and gram negative bacteria.

The process of lipolysis is to release free fatty acids [FFA] and partial glycerides which have either deterimental or beneficial effects in the environment. Lipolytic enzymes have wide range of industrial applications in food, dairy, paper, textile, leather, and detergent industries. These lipolytic enzymes are also used in waste water treatment, production of fine chemicals, pharmaceuticals and cosmetics, synthesis of surfactants and polymers, vegetable fermentation and meat curing.

Due to the wide range of industrial applications, lipases obtained from microorganisms are more interesting as they can be produced with better yields. There are many varieties of catalytic activities that can be used in several applications and the genetic manipulations are easily available. The enzymes are heat stable and involved in the spoilage of a various dairy products (subham verma etal 2014.)

II. MATERIALS AND METHODS:

2.1 Collection of dairy waste water samples:2.2 Isolation of lipolytic bacteria:2.2.1 Preparation of enrichment media:2.2.3 Preparation of confirmatory media:2.3 Screening of lipolytic isolates:3.1 Estimation of lipase activity in the lipolytic bacteria:(a)Effect of pH on lipase activity:(b)Effect of temperature on lipase activity:(c)Effect of different carbon sources on lipase activity:(d)Effect of different nitrogen sources on lipase activity:(e)Effect of different oils as Lipid source on lipase activity:(f) Effect of different metal ions on lipase activity:3.1Optimisation of media parameters:(a)Selection of pH optima for lipase activity:(b)Selection of temperature optima for lipase activity:Fig 2(c)Selection of carbon sources for optimum lipase activity:Fig 3(d)Selection of nitrogen sources for optimum lipase activity:Fig 4(e) Selection of lipid sources as substrates for optimum lipase activity:Fig 5(f) Effect of metal ion on lipase activity:Fig 6Metal ions concentration (1 mM)Concentration (mM)

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