Analysis of the Extracellular Polymeric Substances of Mycobacterium Smegmatis

Bacteria are single-celled organisms comprising soil microfauna and microflora. They are the most numerous soil inhabitants, and reproduce by rapid binary fission under favorable environmental conditions. Different species of bacteria exhibit parasitism, mutualism, and commensalism under varying degrees of association that are obligate, facultative, or symbiotic. In the soil, bacteria survive in the film of moisture surrounding the soil particles. In these conditions, bacteria surviving in the soil water form biotic films called biofilms which affords the bacterial colonies many advantages whilst inhabiting the soil. Bacterial biofilms are a conglomeration of bacteria embedded in a self-synthesized and self-secreted matrix of extracellular polymeric substances (EPS). Serving as a protective hydrated barrier, the EPS provides surface-adhesive properties, facilitates chemical interactions among bacteria, and affords protection from environmental insults including ultraviolet radiation, desiccation and antibiotics. These biofilms have been shown to be able to clean up soil that is contaminated with toxic organics and heavy metals, all of which are potential groundwater pollutants. The main goal of this study is to extract and study the EPS of Mycobacterium smegmatis. By studying the physical and chemical composition of EPS, therapies can be designed to disturb and destroy the EPS, thereby making the task of removing harmful soil bacteria easier. Preliminary studies show that the EPS produced by M. smegmatis is a tangible material capable of withstanding physical and chemical manipulations. Lipids have been extracted using a modified Bligh and Dyer method, and analyzed by TLC. Further investigations include protein and DNA analyses, and biofilm resistance to the antibiotics Streptomycin and Tetracycline.