Microalgae have gained considerable attention as sustainable sources of biomass and valuable metabolites, including lipids. The unicellular green alga Parachlorella kessleri has shown potential for lipid production. In this study, an algae species was isolated from the Yamuna River in Delhi. The organism was cultured and characterized using morphological, biochemical, and molecular approaches, with species identification primarily supported by morphological and taxonomic characterization. This study was performed to examine the influence of media pH, incubation temperature, photoperiod, and CO₂ sparging duration on the growth characteristics of P. kessleri. One-factor-at-a-time (OFAT) approach was used to assess the effect of parameters on cell growth. The lipid content of the organism was also determined. The findings indicated that the maximum biomass production occurred in a 16 day-old culture under the following conditions: pH 8, a temperature of 25 °C, a photoperiod of 16:8 (light:dark), and CO₂ sparging for 2 min twice daily. This species exhibited significant potential for lipid production under normal culture conditions.