skip to main content

Isoflavones and Bioactivities in Over-fermented Tempeh Extracts

1Research Center for Chemistry, National Research and Innovation Agency (BRIN) Republic of Indonesia, Kawasan Puspiptek, Tangerang Selatan 15314, Indonesia

2Department of Food Science and Technology, Faculty of Agricultural Engineering and Technology, IPB University, Bogor 16680, Indonesia

Received: 23 Apr 2021; Revised: 6 Sep 2021; Accepted: 14 Sep 2021; Published: 31 Dec 2021.
Open Access Copyright 2021 Jurnal Kimia Sains dan Aplikasi under

Citation Format:

Tempeh is nutritious food prepared through solid-state fermentation of cooked and dehulled soybeans with Rhizopus sp. for about 48 h. Fermentation beyond 48 h resulted in over-fermented tempeh. There may or may not have been similar research done before, especially related to its antioxidant and cytotoxicity. This study aims to determine the characteristics of fermented tempeh for up to 156 h. Samples were fermented at 0, 24, 48, 60, 72, 84, 96, 108, 120, and 156 h. Samples were dried, grounded, and extracted with acetone, followed by defatting with n‑hexane. Extracts were dissolved in organic solvents for free radical scavenging activity (FRSA) and cytotoxicity assays. The 120-h tempeh extract, at the concentration of 1,000 μg/mL, demonstrated the highest FRSA (81.31% inhibition) against 100 µM 2,2-diphenyl-1-picrylhydrazyl (DPPH) solution. Meanwhile, the 108-h tempeh extract at 1,000 μg/mL possessed the highest cytotoxicity (IC50 of 2.54 μg/mL) against MCF-7 breast cancer cell lines. Liquid Chromatography-Mass Spectrometry/Mass Spectroscopy (LC-MS/MS) analysis revealed the presence of daidzin, genistin, daidzein, and genistein in all extracts. Extracts prepared from 108 h and 120 h tempeh stood out from other extracts in the Partial Least Square (PLS) bi-plot due to their high percentage of inhibition, low response of daidzin, and high responses of the other three isoflavones. The cytotoxicity assays of the standard isoflavones showed that genistein had the lowest IC50 value at 4.82 ± 0.11 μg/mL. Standard genistein showed a low percentage of inhibition at 29.79 ± 9.10.

Fulltext View|Download
Keywords: acetone extract; DPPH; citotoxicity; PLS
Funding: TWAS research grant under contract 15-196 RG/CHE/AS_G-FR3240287020 ; Research Center of Chemistry, Indonesian Institute of Sciences under contract B-036/IPT.2/KP/I/2016

Article Metrics:

  1. J David Owens, Indigenous fermented foods of Southeast Asia, CRC Press, 2014,
  2. Netsanet Shiferaw Terefe, Mary Ann Augustin, Fermentation for tailoring the technological and health related functionality of food products, Critical reviews in food science and nutrition, 60, 17, (2020), 2887-2913
  3. Amadeus Driando Ahnan‐Winarno, Lorraine Cordeiro, Florentinus Gregorius Winarno, John Gibbons, Hang Xiao, Tempeh: A semicentennial review on its health benefits, fermentation, safety, processing, sustainability, and affordability, Comprehensive Reviews in Food Science and Food Safety, 20, 2, (2021), 1717-1767
  4. Reggie Surya, Winiati P Rahayu, Production and characteristics of canned tempe extract, Asian Journal of Food and Agro-Industry, 5, 4, (2012), 299-306
  5. Dwi Larasatie Nur Fibri, Michael Bom Frøst, Indonesian millennial consumers’ perception of tempe–And how it is affected by product information and consumer psychographic traits, Food Quality and Preference, 80, (2020), 103798
  6. Badrut Tamam, Dahrul Syah, Maggy Thenawidjaja Suhartono, Wisnu Ananta Kusuma, Shinjiro Tachibana, Hanifah Nuryani Lioe, Proteomic study of bioactive peptides from tempe, Journal of bioscience and bioengineering, 128, 2, (2019), 241-248
  7. Chen‐Tien Chang, Cheng‐Kuang Hsu, Su‐Tze Chou, Ya‐Chen Chen, Feng‐Sheng Huang, Yun‐Chin Chung, Effect of fermentation time on the antioxidant activities of tempeh prepared from fermented soybean using Rhizopus oligosporus, International Journal of Food Science & Technology, 44, 4, (2009), 799-806
  8. Maciej Kuligowski, Katarzyna Pawłowska, Iwona Jasińska-Kuligowska, Jacek Nowak, Isoflavone composition, polyphenols content and antioxidative activity of soybean seeds during tempeh fermentation, CyTA-Journal of Food, 15, 1, (2017), 27-33
  9. Dian Muzdalifah, Zatil Afrah Athaillah, Anastasia F Devi, Linar Z Udin, Cytotoxicity of ethanol overripe tempe extract against MCF-7 breast cancer cell and its antioxidant activity, AIP Conference Proceedings, 2018
  10. Zatil Afrah Athaillah, Dian Muzdalifah, Asri Lestari, Anastasia Fitria Devi, Linar Zalinar Udin, Nina Artanti, Phenolic compound profile and functionality of aqueous overripe tempe extracts, Current Research in Nutrition and Food Science Journal, 7, 2, (2019), 382-392
  11. Rahayu Utami, Christofora Hanny Wijaya, Hanifah Nuryani Lioe, Taste of water-soluble extracts obtained from over-fermented tempe, International Journal of Food Properties, 19, 9, (2016), 2063-2073
  12. Riris Istighfari Jenie, Sri Handayani, Ratna Asmah Susidarti, Zalinar Udin, Edy Meiyanto, Cytotoxic and Antimetastasis Effect of Ethyl Acetate Fraction from Caesalpinia sappan L. on MCF-7/HER2 Cells, Indonesian Journal of Cancer Chemoprevention, 8, 1, (2017), 42-50
  13. JC De Reu, D Ramdaras, FM Rombouts, MJR Nout, Changes in soya bean lipids during tempe fermentation, Food Chemistry, 50, 2, (1994), 171-175
  14. Chiara Schiraldi, Mario De Rosa, Mesophilic Organisms, in: E. Drioli, L. Giorno (Eds.) Encyclopedia of Membranes, Springer Berlin Heidelberg, Berlin, Heidelberg, 2016,
  15. Indah Dwiatmi Dewijanti, Sofna D Banjarnahor, Triyuliani, Faiza Maryani, Lia Meilawati, Antioxidant activity, phenolic and flavonoids total of ethanolic extract of Ipomoea batata L. leaves (white, yellow, orange, and purple), AIP Conference Proceedings, 2017
  16. Genevieve M Forster, Komal Raina, Ajay Kumar, Sushil Kumar, Rajesh Agarwal, Ming-Hsuan Chen, John E Bauer, Anna M McClung, Elizabeth P Ryan, Rice varietal differences in bioactive bran components for inhibition of colorectal cancer cell growth, Food chemistry, 141, 2, (2013), 1545-1552
  17. Luciane Yuri Yoshiara, Tiago Bervelieri Madeira, Fernanda Delaroza, Josemeyre Bonifácio da Silva, Elza Iouko Ida, Optimization of soy isoflavone extraction with different solvents using the simplex-centroid mixture design, International Journal of Food Sciences and Nutrition, 63, 8, (2012), 978-986
  18. Christian Reichardt, Empirical parameters of solvent polarity as linear free‐energy relationships, Angewandte chemie international edition in English, 18, 2, (1979), 98-110
  19. Hiroshi Murakami, Tomomi Asakawa, Junji Terao, Setsuro Matsushita, Antioxidative stability of tempeh and liberation of isoflavones by fermentation, Agricultural and Biological Chemistry, 48, 12, (1984), 2971-2975
  20. Ritsuko Shirabe, Eiko Saito, Norie Sawada, Junko Ishihara, Ribeka Takachi, Sarah Krull Abe, Taichi Shimazu, Taiki Yamaji, Atsushi Goto, Motoki Iwasaki, Fermented and nonfermented soy foods and the risk of breast cancer in a Japanese population‐based cohort study, Cancer Medicine, 10, 2, (2021), 757-771
  21. Wuryani, A Study of Isoflavones in Soybean and Tempe, University of Reading, Department of Food Science and Technology, 1992
  22. James Devillers, Methods for building QSARs, in: Computational toxicology, Springer, 2013,
  23. Herve Abdi, Lynne J Williams, Partial least squares methods: partial least squares correlation and partial least square regression, in: Computational toxicology, Springer, 2013,
  24. Natnicha Bhumiratana, Mona Wolf, Edgar Chambers IV, Koushik Adhikari, Coffee drinking and emotions: Are there key sensory drivers for emotions?, Beverages, 5, 2, (2019), 27
  25. Livia Dias de Queiros, Amanda Rejane Alves de Ávila, Andressa Vianna Botaro, Danielle Branta Lopes Chirotto, Juliana Alves Macedo, Gabriela Alves Macedo, Combined isoflavones biotransformation increases the bioactive and antioxidant capacity of soymilk, Applied microbiology and biotechnology, 104, 23, (2020), 10019-10031
  26. Chung Hung Lee, Lin Yang, Jin Ze Xu, Sai Ying Venus Yeung, Yu Huang, Zhen-Yu Chen, Relative antioxidant activity of soybean isoflavones and their glycosides, Food chemistry, 90, 4, (2005), 735-741

Last update:

No citation recorded.

Last update:

No citation recorded.