Pakistan Journal of Medical and Health Sciences Vol. 8, Issue 1, JAN – MAR 2014 Website: www.pjmhsonline.com page no.21 Biosurfactant production by Pseudomonas Aeruginosa Strains on 4 ml of inoculum size ABDUL GHANI KORAI 1 , YASER AMEER 2 , SHOAIB ASIF 3 , HAROON HABIB 4 , MUDASER HUSSAIN ABBASI 5 , RANA MUHAMMAD AKHTAR 6 , MIAN ABDUR RASHEED 7 , SALAHUDDIN 8 , AHMED TARIQ 9 , HEENA AWAIS 10 1 Assistant Prof. Forensic medicine & Toxicology, Central Park Medical College, Lahore, 2 Assistant Prof. Forensic medicine & Toxicology, Lahore Medical & Dental College, Lahore 3 Biochemist Scholar , 4 Lecturer Biochemistry 5 Associate Prof. Forensic Medicine & Toxicology 6 Prof. Community Medicine, Avicenna Medical College Lahore , 7 Prof.Forensic Medicine& Toxicology,Shaheed Benazir Bhutto Medical college,Azad Kashmir, 8,9 Lecturer in Forensic Medicine & Toxicology, Physiotherapist Correspondence: Shoaib Asif Email: biofilm@yahoo.com ABSTRACT Aim: To produce biosurfactants from Pseudomonas aeruginosa using agricultural resource and to produce Biosurfactants using low cost materials. Study design: Descriptive study Place and duration of study: Study was conducted at Institute of molecular biology and biotechnology in university of Lahore. Duration of the study was two years. Methods: The volume of sample taken are 4ml, of innoculum from growing culture of Pseudomonas aeruginosa was isolated from contaminated soil collected from industrial area of District Kasoor and flasks were then placed into an orbital shaker at speed of 120rpm. The samples were collected in sterile screw capped bottle, 4-5 cm deep from the soil surface aseptically. The samples were stored at 4 o C till further use. After every 24h the culture broth from each flask was taken to estimate bacterial cell mass. Results:-. surface tension was 64.3, 62.1, 49.8 and 46.4mN/m at time 24, 48, 72 and 96 hours respectively at constant temperature of 37°C and molasses used 0.25g with 4ml inoculum size. The rhamnolipid production was 0.15, 0.4, 0.76 and 0.8 g/L respectively. Similarly the bacterial cell mass was 0.3, 0.33, 0.26 and 0.28 g/L respectively Conclusion: After optimizing various growth and environmental factors a production of rhamnolipid was achieved. Keywords: Biosurfactant, pseudomonas aeruginosa, incolation INTRODUCTION In recent years, much attention has been directed towards biosurfactants owing to their different advantages such as, lower toxicity, higher biodegradability, better environmental compatibility, higher foaming, high selectivity, specific activity at extreme temperatures, pH and salinity, and the ability to be synthesized from renewable feed stocks 1 . Due to their biodegradability, biosurfactants were originally meant to replace chemical surfactants 2 . The choice of inexpensive raw materials is important to the overall economy of the process as they account for 50% of the final production cost and also reduce the expenses with waste treatment 3 . The amphiphiles that form micelles and can be potentially used for surface chemical works are termed surface active agents or surfactants. The enormous market demand for surfactants is currently met by numerous synthetic, mainly petroleum-based chemical surfactants. These compounds are usually toxic to the environment as well as non-biodegradable. They may bio- accumulate and their production, processes and by-products can be environmentally hazardous. It has become important that tightening environmental regulations and increasing awareness for the need to protect the ecosystem have effectively resulted in an increasing interest in biosurfactants as possible