Optimization and Kinetic Studies of Enzymatic Saccharification of Double-Stage Ozonolysis Pretreated Oil Palm Empty Fruit Bunch for Enhanced Sugar Production

Nur Zahidah Abd Majid (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor)
Amnani Shamjuddin (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor)
Mohd Asmadi (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor)
Umi Aisah Asli (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor)
Sharifah Nurain Hussain (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor)
Riyani Tri Yulianti (Chemical Reaction Engineering Group (CREG), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor||Malaysia Research Center for Electronics, National Research and Innovation Agency (BRIN), KST Sama’)
Nur Hidayah Zainan (Centre of Lipid Engineering Applied Research (CLEAR), Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor||Malaysia Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy)
Nardiah Rizwana Jaafar (Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru)
Asiah Nusaibah Masri (UTM-MPRC Institute for Oil & Gas, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia (UTM), 81310 Johor Bahru, Johor||Malaysia Energy Management Group, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia ()
Gwendoline Christophe (Université Clermont Auvergne, CNRS, Clermont auvergne INP, Institut Pascal, 63000 Clermont-Ferrand)
Philippe Michaud (Université Clermont Auvergne, CNRS, Clermont auvergne INP, Institut Pascal, 63000 Clermont-Ferrand)

Bulletin of Chemical Reaction Engineering & Catalysis
2026: BCREC Volume 21 Issue 3 Year 2026 (October 2026)

Oil Palm Empty Fruit Bunch (OPEFB), an abundant lignocellulosic biomass in Malaysia, is a promising feedstock for producing Total Reducing Sugar (TRS), although its recalcitrant structure limits enzymatic conversion. This study proposes a novel double-stage ozonolysis pretreatment integrated with intermediate alkaline swelling to enhance cellulose accessibility and saccharification efficiency. Structural modifications were confirmed through compositional analysis, TGA, XRD, FTIR, and SEM. Enzymatic saccharification (ES) was optimized using Response Surface Methodology (RSM) based on a Face-Centered Central Composite Design (FCCCD), evaluating reaction time, biomass loading, and temperature. Analysis of Variance (ANOVA) indicated a significant model (R2 = 0.88), with optimal conditions of 44 h reaction time, 1.8 % w/v biomass loading, and 50 °C temperature, achieving a maximum TRS yield of 42.75%. The double-stage ozonolysis outperformed single-stage and alkaline pretreatments, yielding the highest cellulose enrichment (up to 79 wt%) and improved digestibility. Kinetic analysis revealed a substantial reduction in the Michaelis-Menten constant ( ) from 175.713 to 9.010 mg/mL, indicating enhanced enzyme–substrate affinity. These findings demonstrate a robust and efficient strategy for improving biomass-to-sugar conversion. Copyright © 2026 by Authors, Published by BCREC Publishing Group. This is an open access article under the CC BY-SA License (https://creativecommons.org/licenses/by-sa/4.0).