In the present investigation bioflocculant production from fungiwas investigated as microbial bioflocculant has advantages over chemical flocculant. They are found to be economical, potential flocculating effects, biodegradability and harmless to humans and the environment. Based on these properties it is proved to be advantageous in many ways and may potentially be applied in production industries for various purposes. In this study, a total of four fungal isolates were obtained and the isolates were screened using screening media for flocculant production, measured using the kaolin solution as a test material. The flocculant activity was measured using UV-Vis spectroscopy and studied for its bioflocculant activity. The functional groups were analysed by Fourier transfer infrared radiation (FTIR). Further, for better quantification of the flocculant production, Response surface methodology (RSM) was used to design an experimental set up which could be used to study the effect of different parameters on the bioflocculant production. © 2015, Sphinx Knowledge House. All rights reserved.