Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 18 3 2026 06 07 439 447 EN Yenni Karlina Department of Pharmaceutical Biology, School of Pharmacy, Bandung Institute of Technology, Bandung, West Java, Indonesia; Department of Pharmaceutical Biology, Faculty of Pharmacy, Jenderal Achmad Yani University, Cimahi, West Java, Indonesia Defri Rizaldy Department of Pharmaceutical Biology, School of Pharmacy, Bandung Institute of Technology, Bandung, West Java, Indonesia Catur Riani Department of Pharmaceutics, School of Pharmacy, Bandung Institute of Technology, Bandung, West Java, Indonesia Fahrauk Faramayuda Department of Pharmaceutical Biology, Faculty of Pharmacy, Jenderal Achmad Yani University, Cimahi, West Java, Indonesia Sukrasno Sukrasno Department of Pharmaceutical Biology, School of Pharmacy, Bandung Institute of Technology, Bandung, West Java, Indonesia 2025 07 24 2026 05 06 Background and Objectives: Enzymes are protein biomolecules that act as catalysts, including cellulase and β-glucosidase with extensive applications. Thus, this work aimed to contrast the activity of both enzymes from tempeh fermented by Rhizopus microsporus at 2 and 7 days of incubation. Materials and Methods: Incubated tempeh was tested for quality evaluation. The crude extracts of cellulase and β-glucosidase were obtained by extracting tempeh with a cold phosphate buffer solution. The presence of the R. microsporus was examined through microscopic identification, and molecular identification using PCR amplification. Cellulase activity was determined using 3.5-dinitrosalicylic acid (DNS) reagent, whereas β-glucosidase activity was evaluated by measuring the release of p-nitrophenol from p-nitrophenyl-β-D-glucopyranoside (p-NPG). Results: The protein content and water content increase with the length of fermentation time. Microscopic identification and molecular identification confirmed the presence of R. microsporus. The highest cellulase activity was found in fresh tempeh (2-day incubation) at 0.0911 U/mL at pH 7, while the highest β-glucosidase activity was found in expired tempeh (7-day incubation) at 0.0042 U/mL at pH 5. Conclusion: These findings indicate that the standard quality contributed to the differences in the enzymatic profile of tempeh incubated for 2 and 7 days. https://ijm.tums.ac.ir/index.php/ijm/article/view/5666 https://ijm.tums.ac.ir/index.php/ijm/article/download/5666/1904