Enhanced catalytic performance of methanol-tolerant Rhizopus stolonifera immobilized on polyurethane foam with hydrophobic coating for biodiesel production

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The limited methanol tolerance and glycerol adsorption tendency of whole-cell catalysts are the main factors contributing to the extended catalytic cycle and limited catalyst reusability in biodiesel preparation. In this study, a 7.5 % methanol-tolerant Rhizopus stolonifera mutant (MTS2) was developed through the atmospheric and room temperature plasma (ARTP), exhibiting a 1.5-fold increase in methanol tolerance compared with the wild type strain (WT-RS), along with a 93.5 % increase in lipases activity. Polyurethane foam PF3 with an average pore size of 0.15 mm and 90 A rigidity is optimal for MTS2 immobilization (MTS2 @PF3), enabling its application in continuous processing for industrial applications. Utilizing soybean oil as a substrate, the catalyst MTS2 @PF3 produced 95.5 % of fatty acid methyl esters (FAME) in 36 hours under optimal conditions (30°C, 180 r/min, catalyst dosage of 15 % g/g oil, 30 % water content, and methanol addition in Scheme 3), achieving over a 50 % reduction in reaction time compared with WT-RS@PF3. To address limitations in catalyst reusability due to glycerol adsorption, various modifiers, including rosin-based modifier (RM), amino silicone oil (ASO), graphene (GRA), betaine and Tween 80 were evaluated. The glycerol adsorption rates experienced substantial reductions (39.3 % with RM and 85.6 % with betaine), and the catalysts maintained above 60 % FAME yield even after 8 reaction cycles. The combination of enhanced methanol tolerance, lipase activity, and reduced glycerol adsorption contribute to enhanced catalytic efficiency and reusability, which can reduce production costs for biodiesel manufacturing. The composite of these attributes makes MTS2 @RM1-PF3 and Betaine-MTS2 @PF3 as promising candidate for industrial biodiesel production, addressing previous limitations in catalyst performance and sustainability. The developed immobilized whole-cell catalysts exhibit promising potential for more sustainable and cost-effective processes in biodiesel manufacturing.