Figure 3. Pareto chart of Placket-Burman Design. The effectiveness of factors was evaluated using two statistical thresholds, namely the Benforrini limit and t-value limit. These thresholds were determined to be 3,898 and 2.306, respectively, for a significance level (α) of 0.05. The Pareto chart visually represents the magnitude of standardized effects through bars. Notably, four bars stand out with the highest values, corresponding to MgSO4, glycerol, glucose, and LB broth concentration.