The biosurfactants produced by microorganisms have high demand from microbial-enhanced oil recovery (MEOR) and they have focused on a chemical surfactant for the past few decades for degrading petro-based pollutants and oil spills due to its non-toxicity and increasing bioavailability. The study aims to identify and screen potential lipopeptide biosurfactant produced by Paenibacillus species employing a design experiment based on RSM. The bacterial culture was isolated from Chilika Lake, India. The data generated from the biosurfactant stability experiments were used to fit a regression model using the parameters such as ph, temp, and salinity to predict the E24 index. R-squared value 0.91 obtained from the ANOVA model explains that the regression model was significant, and the model p-value obtained was < 0.05 and was also statistically significant. Therefore the statistical regression model obtained in the present investigation can predict the E24 index by using any combination of ph, temp, and salinity parameters. The novel isolates obtained in this research were further named Paenibacillus dendritiformis ANSKLAB02 and deposited in GenBank with accession number KU518891. The growth of this species under controlled conditions has a high potential to help in environmental clean-up and is suitable for use in MEOR applications.