VOL.1, NO.1, JULY 2006 ISSN 1990-6145 Journal of Agricultural and Biological Science © 2006 Asian Research Publishing Network (ARPN). All rights reserved. www.arpnjournals.com ANALYSIS OF STAPHYLOCOCCUS SP. FOR SECONDARY METABOLITE PRODUCTION Dr. Nazia Jamil 1 and Nuzhat Ahmed 2 1 Department of Microbiology and Molecular Genetics, University of Punjab, Lahore-54590, Pakistan 2 Centre for Molecular Genetics, University of Karachi, Karachi-75270, Pakistan E-mail: 1 jamil_nazi@yahoo.com ABSTRACT Several samples were collected from a variety of sources such as surface water, deep-sea water, sediments, sea animals and plants from Arabian Sea. The samples were analyzed both qualitatively & quantitatively for the presence of bacteria. Their resistance markers were studied and two Staphylococcus sps. were screened for the production of secondary metabolites. Analytic techniques and electron microscopic observations have revealed the presence of important compounds such as cadmium binding compound, oleic acid, palmatic acid, SMHS1 and 3. These have potential for commercial exploitation. Keywords: Marine bacteria, secondary metabolites, cadmium binding compound. INTRODUCTION The ocean is a vast body of salt water that covers about three-quarters of the earth's surface. The Arabian Sea lies in the northwestern section of the Indian Ocean. Deep water reaches close to the bordering lands except in the northeast, off Pakistan and India. Pakistan spans a remarkable number of the world’s broad ecological regions. These ranges from the coastal areas to the spectacular mountain top. Pakistan has a coast line of 527nautical miles (nm). This coastal area of Pakistan offers a rich repository of marine organisms while microbial flora in these marine environment forms an integral part of this unique ecosystem (Ahmed and Yasmeen, 1988 and Jamil et al., 1999). The marine microorganisms, including bacteria, fungi, and microalgea, have received increasing attention over the past ten years (Davidson, 1995). Jensen and Fenical in 1994 have introduced marine bacteria as a new biomedical source, as well as commenting on both the chemical and ecological perspective of pursuing marine bacteria as a source of new secondary metabolites. Many microbiologists around the world have attempted to isolate bacteria that produce novel products such as antibiotics, enzymes, biologically active substances, polysaccharides, emulsifiers etc. (Horikoshi, 1995).Common sources of metabolite producing bacteria are sea water, sponges, vertebrates, invertebrates, and sea sediments. Examples of such metabolites include Okadanthin, a new C50 carotenoid pigment is produced by a Pseudomonas sp. (Miki et al., 1994). Patel and Hou (1993) have recovered a surfactant, from the sea water isolates of Acinetobacter. This surfactant is used as cleaning agent for oil tankers and oil storage tanks. It is also capable of forming complex with uranium metal, so may be useful for the recovery of uranium from waste material. It has been reported that marine bacteria have interesting genetic markers, such as markers for biodegradation of complex compounds and resistance to heavy metals and antibiotics etc.. Several plasmids which have been isolated from marine bacteria i.e. TOL- palsmid and NAH-Plasmids, are responsible for the production of enzymes which degrade complex compounds like toluene and naphthalene respectively (Saunders, 1977). The genetics of production of few secondary metabolites has been studied such as tetracenomycin (Tcm) C. It is potent inhibitor of the growth of other streptomycetes and has moderate cytotoxicity towards some tumor cells. TcmC production is determined by a cluster of 12 genes contained with in an operon 13kb region of the S. glancescens (Hutchinson, 1992). There is a possibility of creating novel hybrid secondary metabolites by transferring genes from one bacterium to another. The present study was conducted in three phases. In the first phase, after the isolation and purification of bacterial strains, they were characterized for biochemical and genetic characters. In second phase, selected bacterial strains were analyzed for the production of secondary metabolites under different cultural media by using different chromatographic and spectroscopic techniques. Finally the cellular morphology and metabolites was studied by using scanning electron microscopy. MATERIALS AND METHODS Phase-1: Characterization of bacterial strains Sampling sites Samples were collected at four different stations along Karachi coast. These stations were Sandspit (around 25.20 o N 66.8 o E), Clifton (around 24.71 o N 67.17 o E), Lyari outfall (around 47.21 o N 67.10 o E), Manora (around 24.87 o N 66.81 o E), Capemonze (around 24.5 o N, 66.4 o E), Gaddani (around 25.5 o N, 66.7 o E) and Dockyard. 32