In this paper, biomass production parameter optimization was performed using Chlorella medium by design of experiment (DOE) with the help of Qualitek-4 software with bigger is better as quality characteristics with four media components and they are studied at three levels in submerged culture condition. I have studied four parameters like pH, Na2CO3, NH4Cl, and NaNO3 with three levels. These factors are optimized based on their S/N ratios are obtained from Qualitek-4 software and their significant individual interactions, and interactions with each other have been studied. During experimental studies, I had worked on the optimization of several parameters. It has found that there was an enhancement of chlorella pyrenoidose by 6% in chlorella medium