Microbes are being widely applied in various fields like pollution mitigation, biofuel and enzyme production, food industry, agriculture, and medical science. Behavior of microbes toward their environment, especially depends on the microbial cell surface. Biomolecules present on cell surface decide the fluidity or hydrophobicity of microbial cell, which results in the affinity of microbial cell toward the hydrophilic or hydrophobic substance. Microbial cell with higher hydrophobicity has been correlated with cell aggregation, biodegradation of organic pollutants, biofilm production, cell resistance toward hydrophobic substances, etc. Thus, the study of biomolecules that influence cell surface hydrophobicity (CSH) is important for better utilization of these potent natural agents for their different applications. The present study briefs about the different cell surface biomolecules (fatty acids and proteins) that are being reported as responsible for the alteration of CSH, along with the environmental factors that influence microbial cell hydrophobicity.