Study of Mangan Peroxidase (Mnp) Enzymes from Pleurotus Ostreatus Produced using Coffee Grounds and Rice Bran as Substrates through Response Surface Methodology
Abstract
This study aimed to optimize the production of Manganese Peroxidase (MnP) enzyme from Pleurotus ostreatus using agricultural waste substrates such as coffee grounds (CG) and rice bran (RB) through Response Surface Methodology (RSM) approach. Optimization was conducted using a Central Composite Design (CCD) with three main variables, namely MnSO₄ concentration (0.5-1.5) mM, incubation temperature (25-35) °C, and reaction time (20-30) min. The enzyme fractionation process was carried out by ammonium sulfate saturation (0-20%), (20-40%), (40-60%), followed by dialysis, and SDS-PAGE analysis. The results showed that the 0-20% fraction of CG:RB (50:50) substrate had the highest enzyme activity, which was 20.588 ± 0.141 U/mL, with the highest specific activity of 76.967 ± 0.325 U/mg. The highest protein content was found in CG:RB (0:100) substrate at 0.825 ± 0.016 mg/mL. However, the enzyme activity was lower due to the presence of non-enzymatic proteins and natural inhibitors. This study successfully demonstrated that the substrate combination of CG:RB (50:50) provided optimal results for MnP production. The Response Surface Methodology proved effective in optimizing process variables, providing great potential for biotechnology applications using agricultural waste as an environmentally friendly raw material.
Keywords: Pleurotus ostreatus, Manganese Peroxidase, Coffee Dregs, Rice Bran, RSM, SDS-PAGEFull Text:
Full-text PDFReferences
Arunima, Satish, and Satish Verulkar. 2022. “Comparative Analysis of Different Protein Estimation Methods.”The Pharma Innovation JournalSP-11(4): 2091–2095.
Argaw, Biniam, Teklemichael Tesfay, Tesfay Godifey, and Negasi Asres. 2023. “Growth and Yield Performance of Oyster Mushroom (P. Ostreatus (Jacq.: Fr.) Kummer) Using Waste Leaves and Sawdust.” Edited by Maria Serrano.International Journal of Agronomy2023 (October): 1–7. doi:10.1155/2023/8013491.
Blanco. G, Laura, Yolanda Diñeiro, Andrea Díaz-Luis, Ana Coto-Montes, Mamen Oliván, and Verónica Sierra. 2021. “Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat.”Foods10 (5): 1097. doi:10.3390/foods10051097.
Bonaventura. A., and Anna. K., 2022. “Effect of Coffee Grounds as Compost on The Growth of Vorstenlanden Tobacco.” Journal of Global Sustainable Agriculture 2(2): 44-49.
Cahyani. P., Wijanarka., and Budi. R., “Specific Activity of Serratia Marcescens Cellulase By Variation of Amonium Sulfate ((Nh4)2so4) Concentration and Ph.” Journal of Biology 6(2): 41-49.
Chai. W. Y., Umahsreerekah. G. K., Vikineswary. S., And Joash. B. L. T., 2021. “ Assessment Of Coffee Waste In Formulation Of Substrate For Oyster Mushrooms Pleurotus Pulmonarius And Pleurotus Floridanus., Future Foods 4: 100075.
Charis. B., Vianey Ariadna, Eddy Jonatan Moreno-Román, Juan Antonio Vidales Contreras, and Celestino García-Gómez. 2019. “Simultaneous Removal of Cd2+ and Zn2+ from Aqueous Solution Using an Upflow Al-Electrocoagulation Reactor: Optimization by Response Surface Methodology.”Water Science and Technology79 (7): 1297–1308. doi:10.2166/wst.2019.123.
Daugirdas, John T. 2018. “Changes in Total Protein Concentration Due to Fluid Removal During and Shortly after Hemodialysis.”American Journal of Nephrology48 (2): 118–126. doi:10.1159/000491935.
Dimawarnita. F., and Tri. P., 2019. “Activity of Ligninolytic Enzyme of Pleurotus Ostreatus On Media Containing Opefb and Their Application For Dyes Decolorization.” Plantation Tower 87(1): 31-40.
Fadel, Hoda, Rasha Saad, Mohsen Asker, Manal Mahmoud, Shereen Lotfy, Fatma Abd El-Aleem, Mohamed Ahmed, and Eman Elmansy. 2021. “Production of Enzymatically Hydrolyzed Soybean Protein Isolate by a Novel Neutral Protease from Enterobacter Sp. NRCM1 to Be Used as a Base Precursor of Thermal Process Flavors.”Egyptian Journal of Chemistry0 (0): 0–0. doi:10.21608/ejchem.2021.45983.2940.
Gaur. N., Korrapati. N., and Pydisetty., 2018. “Biochemical and kinetic characterization of laccase and manganese peroxidase from novel Klebsiella pneumoniae strains and their application in bioethanol production.” The Royal Society of Chemistry 8 : 15044–15055.
Gwon, Ju-Hui, Hyeok Park, and Ahn-Heum Eom. 2022. “Effect of Temperature, pH, and Media on the Mycelial Growth ofTuber Koreanum.”Mycobiology50 (4): 238–243. doi:10.1080/12298093.2022.2112586.
Hamzah, Pratiwi, Syaifuddin Syaifuddin, Rachmat Rachmat, and Agus Agus. 2022. "Analysis of Mycelium Growth of F1 Seeds of White Oyster Mushrooms (Pleurotus Ostreatus) Using Corn Seeds and Rice Seeds as Media."JASATHP: Journal of Agricultural Products Science and Technology2 (2): 67–77. doi:10.55678/jasathp.v2i2.807.
Heirina, Anna, Rozirwan Rozirwan, and Muhammad Hendri. 2020. "Isolation and antibacterial activity of endophytic fungi in Sonneratia alba mangroves from Tanjung Carat, Banyuasin Regency, South Sumatra."Journal of Science Research22 (1): 16. doi:10.56064/jps.v22i1.562.
Irdawati., and Rada. A., 2024. “Xylanase Specific Enzyme Activity Values of Bacterial Consortium MS 16, MS 18 and MSS 11 at Various Temperatures.” Journal of Tambusai Education 8(2): 28159-28169.
Kohphaisansombat. C., Yanisa. J., Pitak. L., Sukon. T., Jirayut E., Nattawut. R., Charuwan. C., Papichaya. K., Sayanh. S., and Nattawut. B., 2024. “Fabrication Of Mycelium (Oyster Mushroom)-Based Composites Derived From Spent Coffee Grounds With Pineapple Fibre Reinforcement.” MYCOLOGY 15(4): 665–682.
Kumari, Arpana, Balwinder Kaur, Rajendra Srivastava, and Rajender S. Sangwan. 2015. "Isolation and Immobilization of Alkaline Protease on Mesoporous Silica and Mesoporous ZSM-5 Zeolite Materials for Improved Catalytic Properties."Biochemistry and Biophysics Reports2 (July): 108–114. doi:10.1016/j.bbrep.2015.05.009.
Kurien, Biji T., and R. Hal Scofield. 2012. “Accelerated Coomassie Blue Staining and Destaining of SDS-PAGE Gels with Application of Heat.” InProtein Electrophoresis: Methods and Protocols, edited by Biji T. Kurien and R. Hal Scofield, 471–479. Totowa, NJ: Humana Press. doi:10.1007/978-1-61779-821-4_41.
Kuwahara, Masaaki, J.K Glenn, and M.A Morgan. 1984. “Separation and Characterization of Two Extracellular H2O2-Dependent Oxidases from Ligninolytic Cultures of Phanerochaete Chrysosporium.” FEMS Microbiol. Lett., 169, 247-250.
Meza. R, Silvia Leticia, Lázaro Cañizares, Brenda Dannenberg, Betina Bueno Peres, Larissa Alves Rodrigues, Celina Mardade, Maria Antônia De Leon, César Augusto Gaioso, Isabel Egea, and Mauricio De Oliveira. 2024. “Sustainable Rice Bran Protein: Composition, Extraction, Quality Properties and Applications.”Trends in Food Science & Technology145 (March): 104355. doi:10.1016/j.tifs.2024.104355.
Nguyen. D. V., Cham. T. T. D., Chau. N. M. V., Hung. M. N., Tuyet. T. P., Tuyet. M. T. T., And Long. Q. N., 2023. “Data on Chemical Composition of Coffee Husks and Lignin Microparticles as Their Extracted Product.” Data In Brief 51: 109781.
Nurkhotimah., 2017. “The Influence of Temperature and Ph Toward The Activity of The Phosphatase Enzyme Fromthermophilic Bacteria of Gendol River Pasca Merapi Eruption.” Journal of Biology 6(8): 465-471.
Qing, Rui, Shilei Hao, Eva Smorodina, David Jin, Arthur Zalevsky, and Shuguang Zhang. 2022. “Protein Design: From the Aspect of Water Solubility and Stability.”Chemical Reviews122 (18): 14085–14179. doi:10.1021/acs.chemrev.1c00757.
Shen, Chang-Hui. 2019. “Chapter 8 - Quantification and Analysis of Proteins.” InDiagnostic Molecular Biology, edited by Chang-Hui Shen, 187–214. Academic Press. doi:10.1016/B978-0-12-802823-0.00008-0.
Umar, Aisha, and Shakil Ahmed. 2022. “Optimization, Purification and Characterization of Laccase from Ganoderma Leucocontextum along with Its Phylogenetic Relationship.”Scientific Reports12 (1): 2416. doi:10.1038/s41598-022-06111-z.
Wingfield, Paul. 2016. "Protein Precipitation Using Ammonium Sulfate."Current Protocols in Protein Science13 (1). doi:10.1002/0471140864.psa03fs1.
Xue. H. Meng. Y. G., Yan. H. G., Xiao. H. B., and Xuan. W. Z. 2017. “Expression and characteristics of manganese peroxidase from Ganoderma lucidum in Pichia pastoris and its application in the degradation of four dyes and phenol.” BMC Biotechnol 17: 19.
Zhu, Yingpeng, Jiangliu Yu, Chunyan Jiao, Jinfeng Tong, Lei Zhang, Yan Chang, Weina Sun, Qing Jin, and Yongping Cai. 2019. “Optimization of Quercetin Extraction Method in Dendrobium Officinale by Response Surface Methodology.”Heliyon5 (9): e02374. doi:10.1016/j.heliyon.2019.e02374.
Zusfahair, Zusfahair, Dian Riana Ningsih, Dania Putri, and Amin Fatoni. 2018. "Partial Purification and Characterization of Urease from Black-Eyed Pea (Vigna Unguiculata Ssp Unguiculata L.)" 14 (1).
DOI: http://dx.doi.org/10.24845/ijfac.v10.i1.68
Refbacks
- There are currently no refbacks.
Editorial Office:
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sriwijaya
Jl. Palembang-Prabumulih Km.35 Indralaya Ogan Ilir Sumatera Selatan 30662
IJFAC by Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Sriwijaya is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License