Antioxidant activity of a mixture of water-soluble tempeh extract with whey powder that has undergone a Maillard reaction

Antioxidant activity of a mixture of water-soluble tempeh extract with whey powder

  • Azis Boing Sitanggang Department of Food Science and Technology, IPB University, Bogor, Indonesia
  • Nisrina Zulfa Firdausi Department of Food Science and Technology, IPB University, Bogor, Indonesia
  • Slamet Budijanto Department of Food Science and Technology, IPB University, Bogor, Indonesia
Keywords: Antioxidant activity, Maillard reaction, Water-soluble tempeh extract, Whey powder


High amino acid content in tempeh and lactose in whey powder can be combined to perform Maillard reaction. So, it can be used to produce functional food products. This is caused by the formation of Amadori and melanoidin as water-soluble compounds which are reported to have high antioxidant capacity and produce a preferred flavor. The general objective of the study was to obtain an optimum process condition in a mixture of water soluble tempeh extract and whey powder with the highest antioxidant capacity (100 mL). The research method was carried out in three  stages, namely (i) optimation condition of the Maillard reaction between water soluble tempeh extract and whey powder (concentration of whey powder and pH reaction) (ii) optimation of temperature and time for reaction, and also study kinetics of Maillard reactions that occur based on changes in antioxidant capacity, (iii) analysis an optimal mixture of water soluble tempeh extract and whey powder that has undergone a Maillard reaction. The analysis of  the antioxidant activity was carried out using DPPH method, browning intensity, and color analysis using chromameter. The results of the analysis of antioxidant activity showed that the mixture ratio of  water-soluble tempeh extract with whey powder 5 % b/v (100 mL), reacted at 100 °C and pH 5 for 90 minutes can increase the antioxidant activity of the mixture from  67,91 ± 2,62 % to 89,89 ± 0,17 %. That concludes as the optimum condition for Maillard reaction between water-soluble tempeh extract and whey powder.


[AOAC] Association of Official Analytical Chemist. 2012. Official Methods of Analysis. Washington DC (US): AOAC International.
[BSN] Badan Standardisasi Nasional. 2015. SNI 3144:2015 tentang Tempe Kedelai. Jakarta(ID): Badan Standardisasi Nasional.
Adedayo, B.C., Oboh, G., & Akindahusi, A.A. 2010. Changes in the total phenol content and antioxidant properties of pepperfruit (Dennettia tripetala) with ripening. African Journal of Food Science, 4(6), 403-409.
Aimo, J., Promancio, E., & Damiani, P.C. 2016. Determination of reducing sugars in foodstuff applying multivariate second-order calibration. Analytical Methods, 8(23), 4617-4631.
Alonso, S., Rendueles, M., & Diaz, M. 2011. Efficient lactobionic acid production from whey by Pseudomonas taetroloens under pH-shift condition. Bioresource Technology, 102(20), 9730-9736.
Amadou, I., Gbadamosi, O.S., Shi, Y., Kamara, M.T., Jin, S., & Le, G. 2010. Identification of antioxidative peptides from Lactobacillus plantarum Lp6 fermented soybean protein meal. Research Journal of Microbiology, 5(5), 372-380.
Amarowicz, R. 2009. Antioxidant activity of Maillard reaction products. European Journal of Lipid Science and Technology, 111(2), 109-111.
Antony, S.M., Han, L.Y., Rieck, J.R., & Dawson, P.L. 2000. Antioxidative effect of Maillard reaction products formed from honey at different reaction times. Journal of Agricultural and Food Chemistry, 48(9), 3985-3989.
Aryanta, W.R. 2000. Traditional fermented foods in Indonesia (review). Japanese Journal of Lactic Acid Bacteria, 10(2), 90-102.
Astawan, M. 2008. Sehat dengan Tempe Panduan Lengkap Menjaga Kesehatan dengan Tempe. Jakarta(ID): Dian Rakyat.
Astawan, M., Puteri, N.E., & Palupi, N.S. 2017. Karakteristik tepung tempe larut air. Jurnal Pangan, 26(2), 1-12.
Astuti, M., Meliala, A., Dalais, F.S., & Wahlqvist, M.L. 2000. Tempe, a nutritious and healthy food from Indonesia (review). Asia Pasific Journal of Clinical Nutrition, 9(4), 322-325.
Aurand, L.W., Woods, A.E., & Wells, M.R. 1987. Milk and Milk Products. In: Food Composition and Analysis. Dordrecht(NL): Springer.
Babji, A.S., Fatimah, S., Ghassem, M., & Abolhassani, Y. 2010. Protein quality of selcted edible animal and plant protein sources using rat bio-assay. International Food Research Journal, 17, 303-308.
Ballin, N.Z. 2006. Estimation of whey Protein in casein coprecipitate and milk powder by hghperformance liquid chromatography quantification of cysteine. Journal of Agricultural and Food Chemistry, 54, 4131-4135.
Barus, T., Suwanto, A., Wahyudi, A.T., Wijaya, H. 2008. Role of bacteria in tempe bitter taste formation: microbiological and molecular biological analysis based on 16S rRNA gene. Microbiology Indonesia, 2(1), 17-21.
Bavia, A.C.F., Silva, C.E., Ferreira, M.P., Santos Elite, R., Mandarino, J.C.G., & Carrao-Panizzi, M.C. 2012. Chemical composition of tempeh from soybean cultivars specially developed for human consumption. Food Science and Technology, Campinas, 32(3), 613-620.
Budijanto, S., Sitanggang, A.B., & Murdiati, W. 2011. Karakterisasi sifat fisiko-kimia dan fungsional isolat protein biji kecipir (Psophocarpus tetragonolobus L.). Jurnal Teknologi dan Industri Pangan, 22(2), 130-136.
Brand-Williams, W., Cuvelier, M.E., & Berset, C. 1995. Use of a free radical method to evaluate antioxidant activity. LWT-Food Science and Technology, 28(1), 25-30.
Brudzynski, K., & Miotto, D. 2011. Honey melanoidins: Analysis of the compositions of the high molecular weight melanoidins exhibiting radical-scavenging activity. Food Chemistry, 127, 1023-1030.
Campbell-platt G. 1994. Fermentedf foods - a world perspective. Food Research International, 27, 25-257.
Chang, C., Hsu, C., Chou, S., Chen, Y., Huang, F., & Chung, Y. 2009. 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), 799-806.
Chawla, S.P., Chander, R., & Sharma, A. 2007. Antioxidant formation by gammairradiation of glucose-amino acid model systems. Food Chemistry, 103(4), 1297 -1304.
Chegeni, G., & Taheri, M. 2013. whey powder: process technology and physical properties: a review. Middle-East Journal of Scientific Research, 13(10), 1377-1387.
Chen, C.C., Shih, Y.C., Chiou, P.W.S., & Yu, B. 2010. Evaluating nutritional quality of single stage- and two stage-fermented soybean meal. Asian-Australasian Journal of Animal Sciences, 23(5), 598–606.
Fardiaz, D., Dedin, F.R., Apriyantono, A., & Andarwulan, N. 2006. Isolasi dan karakterisasi melanoidin kecap manis dan peranannya sebagai antioksidan. Jurnal Teknologi dan Industri Pangan, 17(3), 204-213.
Fibri, D.L.N., & Frøst, M.B. 2019. Indonesian millennial consumer’s perception of tempe and how it is affected by product information and consumer psychographic traits. Food Quality and Preference, 80, 1-32.
Gibbs, B.F., Zougman, A., Masse, R., & Mulligan, C. 2004. Production and characterization of bioactive peptides from soy hydrolysate and soy-fermented food. Food Research International, 37(2),123-131.
Guo, Y., Pan, D., & Tanokura, M. 2009. Optimisation of hydrolysis conditions for the production of the angiotensin-I converting enzyme (ACE) inhibitory peptides from whey protein using response surface methodology. Food Chemistry, 114, 328 – 333.
Han, L., Li, F., Yu, Q., & Li, D. 2017. In vitro antioxidant and cytoprotective properties of Maillard reaction products from phloridzin-amino acid model systems. Journal of the Science of Food and Agriculture, 98(2), 590-597.
Haron, H., & Raob, N. 2014. Nutrition and food changes in macronutrient, total phenolic and anti-nutrient contents during preparation of tempeh. Journal of Nutrition & Food Sciences, 4(2), 1-5.
Hashiba, H. 1982. The browning reaction of amadori compounds derived from various sugars. Agricultural and Biological Chemistry, 46(2), 547-548.
Herawati, D.A., & Wibawa, D.A.A. 2011. Pengaruh konsentrasi susu skim dan waktu fermentasi terhadap hasil pembuatan soyghurt. Jurnal Ilmiah Teknik Lingkungan, 1(2), 48-58.
Hodge, J.E. 1953. Chemistry of browning reactions in model systems. Journal of Agricultural and Food Chemistry, 1(15), 928–943.
Hull, P. 2010. Glucose Syrups: Technology and Applications. Chichester(UK): John Wiley & Sons, Ltd.
Hwang, I.G., Kim, H.Y., Woo, K.S., Lee, J., & Jeong, H.S. 2011. Biological activities in sugar amino acid model system. Food Chemistry, 126(1), 221-227.
Illanes, A. 2011. Whey upgrading by enzyme biocatalysis. Electronic Journal of Biotechnology, 14(6), 1–28.
Jing, H., & Kitts, D.D. 2004. Antioxidant activity of sugar-lysine Maillard reaction products in cell free and cell culture systems. Archives of Biochemistry and Biophysics, 429(2), 154-163.
Karseno, Erminawati, Yanto, T., Setyowati, R., & Haryanti, P. 2018. Effect of pH and temperature on browning intensity of coconut sugar and its antioxidant activity. Food Research, 2(1), 32-38.
Kucükboyaci, N., Ilginer, Ö., Torul, H., Tamer, U., Adigüzel, N., Banİ, B., & Köroğlu, A. 2014. Quantitative determination of isoflavones by HPLC-UV method and antioxidant activity of Trifolium longidentatum. Turkey Journal of Pharmacy Science, 11(2), 185-194.
Kuligowski, M., Pawłowska, K., Kuligowska, I.J., & Nowak, J. 2017. Isoflavone composition, polyphenols content and antioxidative activity of soybean seeds durin g tempeh fermentation. CyTA - Journal of Food, 15(1), 27-33.
Kiers, J., Van laeken, A.E.A., Rombouts, F., & Nout, M.J. 2000. In vitro digestibility of Bacillus fermented soya bean. International Journal of Food Microbiology, 60(2-3), 163–169.
Kiers, J.L., Meijer, J.C., Nout, M.J.R., Rombouts, F.M., Nabuurs, M.J.A., & van der Meulen, J. 2003. Effect of fermented soya beans on diarrhoea and feed efficiency in weaned piglets. Journal of Applied Microbiology, 95(3), 545–552.
Kustyawati, M.E. 2009. Kajian peran yeast dalam pembuatan tempe. Agritech, 29(2), 64-70.
Kustyawati, M.E., Nawansih, O., & Nurdjanah, S. 2017. Profile of aroma compounds and acceptability of modified tempeh. International Food Research Journal, 24(2), 734-740.
Laidler, K.J. 1987. Analysis of kinetic results (3rd ed.). Chemical kinetics. New York(US): Harper & Row Publisher.
Liu, C.F., & Pan, T.M. 2011. Beneficial effects of bioactive peptides derived from soybean on human health and their production by genetic engineering. IntechOpen. 311-328.
Liu, S.C., Yang, D.J., Jin, S.Y., Hsu, C.H., & Chen, S.L. 2008. Kinetics of color development, pH decreasing, and anti-oxidative activity reduction of Maillard reaction in galactose/glycine model systems. Food Chemistry. 108:533-541.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., & Randall, R.J. 1951. Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275.
Maillard, M.N., Billaud, C., Chow, Y.N., Ordonaud, C., & Nicholas, J. 2007. Free radical scavenging of polyphenoloxidase activity and copper chelating properties of model Maillard systems. LWT - Food Science and Technology, 40, 1434-1444.
Motarjemi, Y. 2002. Impact of scale fermentation technology on food safety in developing countries. International Journal of Food Technologgy, 75, 213-229.
Müller, V. 2008. Bacterial fermentation. In: Encyclopedia of Life Sciences (ELS). Chichester(UK): John Wiley & Sons, Ltd.
Mulyowidarso, R.K., Fleet, G.H., & Buckle, K.A. 1990. Association of bacteria with the fungal fermentation of soybean tempe. Journal of Applied Bacteriology, 68, 43-47.
Mursyid, Astawan, M., Muchtadi, D., Wresdiyati, T., Widowati, S., Bintari, S.H., & Suwarno, M. 2014. Evaluasi nilai gizi protein tepung tempe yang terbuat dari varietas kedelai impor dan lokal. Jurnal Pangan, 23(1), 33-42.
Mshayisa, V.V. 2016. Antioxidant effects of Maillard reaction products (MRPs) derived from glucose-casein model systems [thesis]. Bellville(ZA): Faculty of Applied Science, Cape Peninsula University of Technology.
Nooshkam, M., Varidi, M., & Bashash, M. 2019. The Maillard reaction products as food-born antioxidant and antibrowning agents in model and real food systems. Food Chemistry, 275, 644-660.
Nout, M.J.R., & Kiers, J.L. 2005. Tempe fermentation, innovation and functionality: update into the third millenium. Journal of Applied Microbiology, 98(4), 789-805.
Nurdini, A.L., Nuraida, L., Suwanto, A. & Suliantari. 2015. Microbial growth dynamics during tempe fermentation in two different home industries. International Food Research Journal, 22(4), 1668-1674.
O, H.B., Song, K.Y., Joung, K.Y., Shin, S.Y., & Kim, Y.S. 2019. Effects of chia (salvia hispanica L.) seed roasting conditions on quality of cookies. Italian Journal of Food Science, 31(1), 54-66.
Osada, Y., & Shibamoto, T. 2006. Antioxidative activity of volatile extracts from Maillard model systems. Food Chemistry, 98(3), 522 – 528.
Panesar, P.S., & Kennedy, J.F. 2012. Biotechnological approaches for the value addition of whey. Journal Critical Reviews in Biotechnology, 32(4), 327-348.
Pradana, F.R., Anwar, C., Fridayani, N., Aziz, H.A., & Assyfa, A.N. 2017. Inovasi minuman sehat berbasis whey dan sari buah tropis. Asian Journal of Innovation and Entrepreneurship, 2(3), 239–246.
Prazeres, A.R., Carvalho, F., & Rivas, J. 2012. Cheese whey management: a review. Journal of Environmental Management, 110, 48–68.
Prinyawiwatkul, W., Eitenmiller, R.R., Beuchat, L.R., McWatters, K.H., & Phillips, R.D. 1996. Cowpea flflour vitamins and trypsin inhibitor affected by treatment and fermentation with Rhizopus microsporus. Journal of Food Science, 61, 1039-1042.
Puteri, N.E., Astawan, M., Palupi, N.S., Wresdiyati, T., & Takahi, Y. 2018. Characterization of biochemical and functional properties of water-soluble tempe flour. Food Science and Technology, Campinas, 38(1), 147-153.
Rusdah, R., Suhartono, T., Palupi, N.S., & Ogawa. 2017. Tingkat kelarutan peptida tempe dengan bobot molekul kecil pada berbagai jenis pelarut. Agritech, 37(3), 327-333.
Sanjukta, S., & Rai, A.K. 2016. Review: Production of bioactive peptides during soybean fermentation and their potential health benefits. Trends in Food Science & Technology, 50, 1-10.
Schaafsma, G. 2000. The protein digestibility corrected amino acid score. Journal of Nutrition, 130(7), 1865S-1867S.
Seumahu, C.A., Suwanto, A., Rusmana, I., & Solihin, D.D. 2013 Bacterial and fungal communities in tempe as reveal by amplifified ribosomal intergenic sequence analysis. HAYATI Journal of Bioscience, 20(2), 65-71.
Sitanggang, A.B., Drews, A., & Kraume, M. 2016. Recent advances on prebiotic lactulose production. World Journal of Microbiology and Biotechnology, 32(9), 154-163.
Sitanggang, A.B., Lesmana, M., & Budijanto, S. 2020a. Membrane-based preparative methods and bioactivities mapping of tempe-based peptides. Food Chemistry, 127193.
Sitanggang, A.B., Sinaga, W.S.L., Wie, F., Fernando, F ., & Krusong, W. 2020b. Enhanced antioxidant activity of okara through solid state fermentation of GRAS Fungi. Food Science and Technology, 40(1), 178-186.
Soares, L.L., Lucas, A.M.M., & Boaventura, G.T. 2005. Can organic and transgenic soy be used as a substitute for animal protein by rats?. Brazilian Journal of Medical and Biological Research, 38(4), 583-586.
Steinkraus, K.H. 1983. Traditional food fermentations as industrial resources. Acta Biotechnology, 3(1), 3-12.
Suwanto, A., Rahayu, G., & Nuraida, L. 2013. Population dynamics of yeasts and lactic acid bacteria (LAB) during tempe production. HAYATI Journal of Bioscience, 20(2), 57-64.
Tamam, B., Syah, D., Suhartono, M.T., Kusuma, W.A., Tachibana, S., & Lioe, H.N. 2019. Proteomic study of bioactive peptides from tempe. Journal of Bioscience and Bioengineering, 128(2), 241-248.
Wagner, K.H., Derkits, S., Herr, M., Schuh, W., & Elmadfa, I. 2002. Antioxidant potential of melanoidins isolated from a roasted glucose– glycine model. Food Chemistry, 78, 375–382.
Wilson, S., Blaschek, K., & Mejia, E.G. 2005. Allergenic proteins in soybean: processing and reduction of P34 allergenicity. Nutrition Reviews, 63(2), 47-58.
Yoon, D.K., Han, K.J., Won, Y.C., Da, S.J., Ha, J.L., Jung, K.H., & Chi, H.L. 2018. Effect of Allium hookeri root on physicochemical, lipid, and protein oxidation of Longissimus Dorsimuscle meatball. Korean J. Food Sci. Anim. Resources, 38(6), 1203-1212.
Yu, X., Zhao, M.Y., Hu, J., Zeng, S.T., & Bai, X.L. 2012. Correspondence analysis of antioxidant activity and UV-Vis absorbance of Maillard reaction products as related to reactans. LWT - Food Science and Technology, 46, 1-9.
Zayas, J.F. 2012. Functionality of proteins in food. New York(US): Springer.
Zeng, Y., Zhang, X., Guan, Y., & Sun, Y. 2011. Characteristics and antioxidant activity of Maillard Reaction products from psicose-lysine and fructose-lysine model systems. Journal of Food Science, 76, 398-403.
How to Cite
Sitanggang, A. B., Firdausi, N. Z., & Budijanto, S. (2021). Antioxidant activity of a mixture of water-soluble tempeh extract with whey powder that has undergone a Maillard reaction: Antioxidant activity of a mixture of water-soluble tempeh extract with whey powder. Future of Food: Journal on Food, Agriculture and Society, 9(5). Retrieved from
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