Microbial biofilm is a microbial assemblage which is formed by bacterial adhesion, growth and expansion, enclosed in a self-produced polymeric matrix that is adherent to an inert or living surface. Biofilms are group or microorganisms in which microbes produced extracellular polymeric substances (EPS), such as polysaccharides, proteins and extracellular microbial DNA. The biofilm can consist of one or more microbial (bacterial or fungal) species and formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment, especially in the hostile environment. Bacterial biofilms are normally beyond the access of antibiotics and human immune system and antibiotic treatment is cu rrently most effective mea sure for the control of microbial infections. However, antibiotic treatments are almost impossible to remove biofilm infections as the pathogenic bacteria in biofilms are resistant to current therapeutic regimes due to their resistant phenotype. The efficient eradication of biofilm is major concern in healthcare sector, especially in living system where use of harsh chemicals and high temperature are unthinkable. However, milder reagents such as enzymes can be of great help as their actions are highly specific to target molecule and have capability to disrupt the structural integrity of the biofilm matrix. The degradation of extra polymeric substance exposes the pathogenic bacterial cells to antibiotics, which along with host immune response acts more efficiently to clear the infectious agents. The major enzymes used to degrade biofilm are alginate lyase, DNase I, -amylase, protease and dispersin B. The knowledge of chemical nature of the EPS in biofilm helps in deciding that requirement for the use of single enzyme or combination of various enzymes for efficient dispersion of microbial biofilms.