Qualitative Screening and Activity Assay of L-asparaginase-Producing Fungi from Tempeh
DOI:
https://doi.org/10.33394/bioscientist.v13i4.18443Keywords:
Fungi, L-asparaginase, screening, tempehAbstract
This study aims to isolate and determine the types of fungi in tempeh that are capable of producing the enzyme L-asparaginase and how the enzyme activity is produced. The following steps were taken: Fungi were isolated using the pour plate method with Potato Dextrose Agar (PDA) medium; morphological characterization of fungi was performed macroscopically and microscopically.The ability to produce asparaginase was assessed qualitatively using modified liquid M9 medium and quantitative assay of L-asparaginase activity with the Nessler reagent. Potential fungi were identified based on identification books. The results of fungal isolation from tempeh yielded of 7 fungal isolates. Qualitative selection of fungal isolates from tempeh capable of producing L-asparaginase resulted in 4 fungi capable of producing the L-asparaginase enzyme, indicated by a pink color change in the medium. Quantitative assay was performed using an enzyme activity test .The results of quantitative selection showed that the highest activity of L-asparaginase enzyme was found in fungal isolate with the code TP 3, reaching 16,386 U/mL, which has been identified macroscopically and microscopically and is classified as belonging to the genus Aspergillus. These findings indicate that fungi from tempeh have the potential to produce the enzyme L-asparaginase.
References
Adimas, M. A., Abera, B. D., Adimas, Z. T., Woldemariam, H. W., & Delele, M. A. (2024). Traditional food processing and acrylamide formation: A review. Heliyon, 10(9), e30258. https://doi.org/10.1016/j.heliyon.2024.e30258
Adhikari, S., & R., M. (2017). Use of microbial asparaginase to mitigate acrylamide formation in fried food. Food Science & Nutrition Technology, 2(2), 1–6.
Ambreen A, Zia MA, Asgher M, Muhammad F. (2019) .Purification of robust L-Asparaginase from Capsicum annum and its therapeutic efficacy in Leukemia. Pakistan Veterinary Journal 39(3).
Al Zubairy, M. A., Thamer, F. H., Thabet, H., & Aldeen, K. S. (n.d.). (2023). Production of L-Asparaginase by Yemeni Filamentous Fungi. Archieves of Biotechnology and Medicine.
Atika, F. N. & Susanto, A. (2019). Identifikasi Rhizopus sp dan Aspergillus sp pada tempe yang tersimpan dalam suhu ruang. Jurnal Insan Cendekia.
Dewi, L., Paramastuti, R., & Rizkaprilisa, W. (2024). Tempe “Super Food” Untuk Kalangan AnakAnak Tk Di Salatiga Dan Semarang. Communnity Development Journal, 5(3), 5419–5426. https://doi.org/10.31004/cdj.v5i3.27136
Basha, N.S., Rekha, R., Komala, M., dan Ruby, S.. (2009). Production of Extracellular Anti-Lukaeia Enzyme L-asparaginase from arine actinomycetes by Solid-State and Submerged Fermentation : Purification and Characterization.Tropical Journal of Pharmaceutical Research, 8(4), 353-360.
Blumenthal, C. Z. (2004). Production of toxic metabolites in Aspergillus niger, Aspergillus oryzae, and Trichoderma reesei: Justification of mycotoxin testing in food grade enzyme preparations derived from the three fungi. Regulatory Toxicology and Pharmacology, 39(2), 214–228. https://doi.org/10.1016/j.yrtph.2003.09.002.
Badoei-Dalfard, A. (2016). L-asparaginase production in the Pseudomonas pseudoalcaligenes strain JHS-71 isolated from Jooshan hot spring. Molecular Biology Research Communications, 5(1), 1–10. https://pubmed.ncbi.nlm.nih.gov/27844015
Cachumba, J. J. M., Antunes, F. A. F., Peres, G. F. D., Brumano, L.P., Santos, J. C. D., and Silva, S. S. D., (2016). Current Applications and Different Approaches for Microbial L-asparaginase Production. Brazilian Journal of Microbiology 47, 77–85. https://doi.org/10.1016/j.bjm.2016.10.004.
Dange, V. U., & Peshwe, S. A. (2011). Production, purification and characterization of fungal L-asparaginase. Bionano Frontier, 4(1), 162–167.
Dinarvand, Behnus, Rezaei, Parisa Fathi, Akbari, Neda. (2020). Optimization of L-asparaginase Production from a Lactobacillus sp. Isolat from Traditional Dairy Products. Biological Journal of Microorganisms. No.34.
Ekpenyong, M., Asitok, A., Antigha, R., Ogarekpe, N., Ekong, U., Asuquo, M., Essien, J., & Antai, S. (2021). Bioprocess optimization of nutritional parameters for enhanced anti-leukemic L-asparaginase production by Aspergillus candidus UCCM 00117: A sequential statistical approach. International Journal of Peptide Research and Therapeutics, 27(3), 1501–1527. https://doi.org/10.1007/s10989-021-10188-x
Freitas, M., Souza, P., Cardoso, S., Cruvinel, K., Abrunhosa, L. S., Ferreira Filho, E. X.,& O. Magalhães, P. (2021). Filamentous fungi producing L-asparaginase with low glutaminase activity isolated from Brazilian Savanna soil. Pharmaceutics, 13(8), 1268.
Han, S., Jung, J., & Park, W. (2014). Biochemical characterization of L-asparaginase in NaCl-tolerant Staphylococcus sp. OJ82 isolated from fermented seafood. Journal of Microbiology and Biotechnology, 24(8), 1096–1104. https://doi.org/10.4014/jmb.1405.0502
Hartanti, A. T., Rahayu, G., & Hidayat, I. (2015). Rhizopus species from fresh tempeh collected from several regions in Indonesia. HAYATI Journal of Biosciences, 22(3), 136–142. https://doi.org/10.4308/hjb.22.3.136
Imada, A., Igarasi, S., Nakahama, K., & Isono, M. (1973). Asparaginase and glutaminase activities of micro-organisms. Journal of General Microbiology, 76(1), 85–99. https://doi.org/10.1099/00221287-76-1-85
Jia, R., Wan, X., Geng, X., Xue, D., Xie, Z., & Chen, C. (2021). Microbial L-asparaginase for application in acrylamide mitigation from food: Current research status and future perspectives. Microorganisms, 9(8), 1659.
Juliasih, N. L. G. R., Ratri, L. M., Setyawan, A., Rilyanti, M., Rinawati, R., Kiswandono, A. A., & Kurniawati, F. (2024). Nessler Method Verification for Determining Ammonia in Shrimp Pond Wastewater and Its Application in the Ammonia Adsorption Test with Lampung Natural Zeolite. Alchemy Journal Research, 20(2), 257. https://doi.org/10.20961/alchemy.20.2.72456.257-266.
Othman, S. I., Mekawey, A. A., El Metwally, M. M., Gabr, S. A., Alwaele, M. A., Al Fassam, H., & Saber, W. I. (2022). Rhizopus oryzae AM16; a new hyperactive L asparaginase producer: Semi solid state production and anticancer activity of the partially purified protein. Biomedical Reports, 16(3), 1-9.
Pitt, J. I., & Hocking, A. D. (2009). Fungi and Food Spoilage. Springer.
Prativi, A., Saleky, Y. W., Suparman, S., Hapsari, A. I., Mulyo, G., & Agung, F. (2023). Pengembangan formula enteral tinggi BCAA (branched-chain amino acids) berbahan dasar tepung tempe (Rhizopus oryzae) dan whey protein isolate untuk pasien dengan sirosis hati. Jurnal Gizi dan Dietetik, 4(1). https://doi.org/10.34011/jgd.v4i1.2724
Oktavia, A., Zainal, Z., Djalal, M., Hidayat, S. H., & Azkiyah, M. (2024). Exploring the utilization of fungi in Indonesian traditional foods: A review. BIO Web of Conferences, 96, 01025. https://doi.org/10.1051/bioconf/20249601025.
Osama, S., El-Khawaga, M.A., El-Hawary, S., Abou-Zaid, A.A. & El-Din, M.I. (2023) Optimization and characterization of antileukemic L-asparaginase produced by Fusarium solani endophyte. AMB Express, 13(1), 96. https://doi.org/10.1186/s13568-023-01602-2
Sine, Y., & Soetarto, E. S. (2018). Isolasi dan identifikasi kapang Rhizopus pada tempe gude (Cajanus cajan L.). Savana Cendana, 3(04), 67–68. https://doi.org/10.32938/sc.v3i04.487.
Utari, D. M., Rimbawan, R., Riyadi, H., Muhilal, & Purwantyastuti. (2010). Pengaruh pengolahan kedelai menjadi tempe dan pemasakan tempe terhadap kadar isoflavon (Effects of soybean processing becoming tempeh and the cooking of tempeh on isoflavones level). Penelitian Gizi dan Makanan (PGM), 33(2), 148–153.
Yuliana, R., & Qurrohman, M. T. (2022). Pengaruh variasi konsentrasi sari pati buah sukun sebagai alternatif media semi sintetik pada pertumbuhan jamur Candida albicans. Journal of Indonesian Medical Laboratory and Science (JoIMedLabS), 3(1), 65–79. https://doi.org/10.53699/joimedlabs.v3i1.67
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Niva Tahyat, Yurnaliza, Liana Dwihastuty
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
