SCREENING AND IDENTIFICATION OF BACILLUS SPP.LIPASE PRODUCING BACTERIA FROM LIPID CONTENT OF KITCHEN WASTES

* Sher Singh Gill 1 , Archana Shrivastav 1 and Asha Mukul Jana 2 . 1. College of Life Sciences, Cancer Hospital and Research Institute, campus, Gwalior, MP, India. 2. Retired Scientist, DRDE, Gwalior, MP, India. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History

. Bacterial enzymes are more preferred over fungal enzymes because of their higher activities and neutral or alkaline pH optima. In order to increase the cell yields and the enzymatic activities of the cells, genetic and environmental manipulations can be performed more readily on bacterial cells due to their short generation times, their simple nutritional needs and easy screening procedures for desired properties (Hasan, F., et al., (2006).
Growth conditions affect the synthesis of lipase by microorganisms. Carbon and nitrogen sources, the presence of activators and inhibitors, incubation temperature, pH, inoculum amount and oxygen tension can influence lipase production (Gupta, R., et al., 2004). The aim of present study is to isolate and characterize bacterial strains from lipids containing kitchen waste by determining their ability to produce lipase enzymes and their efficacy of effective bacteria use in the degradation of kitchen waste.

ISSN: 2320-5407
Int. J. Adv. Res. 4 (9), 2319-2322 2320 Materials and Methods:-Sample collection:-Lipids and oil content of kitchen waste were collected in plastic bags from Cafeteria house at near College of Life sciences, CHRI hill campus, Gwalior in Madhya Pradesh, India. The isolation of bacterial isolate was carried out by serially diluting the kitchen waste as samples in sterile water and subsequently plating on the Nutrient agar medium.

Isolation and screening of lipase producing bacterial isolates:-
In the present study, lipids contain kitchen waste as sample was collected from a Cafeteria nearby College of Life Sciences, CHRI, Hill campus, Gwalior in a sterile container for the isolation of lipase producing bacteria under laboratory condition. For the isolation of lipolytic bacteria, 1.0 gm of sample was dissolved in 100 ml of distilled water. It was then serially diluted (10 -1 to 10 -6 ) and the diluted samples were inoculated on tributyrin agar for screening of lipase producing bacteria. And inoculated petriplates were incubated at 37 0 C for 24 to 48 hours and the formation of clear zone around the colonies on the plate was considered as lipolytic microbes.

Isolation and Screening of lipase producing bacterial isolates:-
The isolation of bacterial isolate were carried out by serially diluting the volatile samples in sterile water and subsequently plating on the 1.5% Tributyrin agar base medium by using pour plate method. The plates were incubated at 37"Cfor 24-48hrs. And observe for zone formation (Fig.-01). A clear zone around the colonies indicates the production of lipase (Cardenas, J., et al, 2001). The lipase positive bacterial strains were further purified, grown in nutrient broth at 37"C for 24 hrs and screened for the ability to produce lipase with tributyrin, the bacterial isolates L1, and L2. During lipase activities, more lipase production by L1 bacterial strains than the L2 bacterial strain (Fig.-02). These bacteria was identified and further studies were carried out by morphological, physiological and biochemical characteristics (Table-01  Enzyme assay of lipase producing bacterial isolates-According to lipase activity of L1 and L2 bacterial strains are given at 24hrs of incubation at 37 0 C, the Bacillus cereus (L1) showed a maximum produced amount recorded as 55.0 U/ml, whereas the minimum production observed at 37 0 C, at 24hrs and the amount was recorded as 15.0 U/ml by Bacillus megaterium (Fig-02).