<p>Recently, valorization of waste carbon-rich industrial streams to bio-based chemicals has taken center stage. Microbial production of 1, 3-PDO via glycerol route has resurrected owing to the booming biodiesel industry. The present study aimed to evaluate fed-batch fermentation for enhancing 1, 3-PDO production from a novel alkalophilic glycerol assimilating <i>Citrobacter freundii</i> IIPDR3, under shake flask conditions. Simultaneously, the feasibility of substituting pure glycerol (PG) with pretreated crude glycerol (PCG) was explored. Paired T-test revealed that increasing PG concentration (50–80 gL⁻¹) under fed batch conditions, displayed significant positive correlation with 1, 3-PDO titers (ρ: 0.982; α:0.005) and glycerol uptake rate (GUR) (ρ: 0.874; α:0.003).With PCG as substrate, shifting from batch to fed-batch radically enhanced the GUR and 1, 3-PDO titers by 166.1% and 281.7%, respectively. This effect was more pronounced when fed-batch was conducted with 40 gL⁻¹ rather than 30 gL⁻¹ initial PCG concentration and increasing inoculum size. With PCG (at 60gL⁻¹), the best titer, yield, and productivity (TYP) matrices of 1, 3-PDO attained was: 22.03 gL⁻¹, 0.37gg⁻¹, and 1.46gL⁻¹&#xa0;h⁻¹. Additionally, high L ( +) lactic acid titers were achieved by switching from batch to fed-batch fermentation. With PG and PCG, its highest titers were 15.4 gL⁻¹, and 12.4 gL⁻¹, respectively.</p> Graphical Abstract <p></p>