Email this article 
Utilization of Partially Purified Papain Enzyme in Mallika Black Soybean Tempeh Hydrolysate as Umami Seasoning
Abstract
Tempeh made from Mallika black soybean (Glycine max (L.) Merr. var. Mallika) can be fermented for up to 4 days and can be further optimized by adding partially purified papain enzyme obtained from California variety papaya leaves (Carica papaya (L.) var. California). Enzyme can be added to the hydrolysates to degrade protein into short-chain peptides and free amino acids, contributing to umami taste sensory attributes. The study aimed to determine the best ammonium sulfate fractionation of crude papain enzyme and the best physicochemical characteristics of black soybean tempeh protein hydrolysate. The addition of ammonium sulfate fractionation used was 0% to 80%; fermentation time was 2 to 4 days; and the concentration of enzyme added was 0%(w/v) to 1.5%(w/v). The results showed that the 40% fractioned papain enzyme gave the highest protease activity value (0.98±0.04 U ml-1) and most of the papain enzyme was precipitated in this fraction leaving impurities. The black soybean tempeh hydrolysates with 4 × 1% showed the best physicochemical characteristic because it produced the highest umami substance. The best characteristics were moisture content (17.97±0.46%), glutamic acid content (171.58±5.72 mg g-1) that was caused by a transamination reaction, dissolved protein content (470.66±19.50 mg g-1), degree of hydrolysis (43.64±1.99%) and lightness (46.02±0.97). The umami substance’s amino acids are high in content, such as glutamic and aspartic acids (59.89±0.31 mg g-1 and 26.47±0.09 mg g-1). Sensory evaluation showed that treatment 4 × 1% demonstrated no significant difference in umami intensity with MSG (monosodium glutamate).
Keywords
Full Text:
PDFReferences
Adawiyah, D. R., & Setiawan, F. (2017). Ambang deteksi dan preferensi rasa umami dalam model pangan. Jurnal Teknologi dan Industri Pangan, 28(1), 55–61. https://doi.org/10.6066/jtip.2017.28.1.55
Afifah, D. N., Rahma, A., Nuryandari, S. S., Alviche, L., Hartono, P. I., Kurniawati, D. M., Wijayanti, H. S., Fitranti, D. Y., & Purwanti, R. (2019). Nutrition content, protein quality, and antioxidant activity of various tempeh gembus preparations. Journal of Food and Nutrition Research, 7(8), 605–612. https://doi.org/10.12691/jfnr-7-8-8
Airaodion, A. I. (2019). Toxicological effect of monosodium glutamate in seasonings on human health. Global Journal of Nutrition & Food Science, 1(5), 1–9. https://doi.org/10.33552/gjnfs.2019.01.000522
Alahmad, K., Xia, W., Jiang, Q., & Xu, Y. (2022). Effect of the degree of hydrolysis on nutritional, functional, and morphological characteristics of protein hydrolysate produced from bighead carp (Hypophthalmichthys nobilis) using ficin enzyme. Foods, 11(9), 1320. https://doi.org/10.3390/foods11091320
Amer, H. E. A. (2019). Purification of proteins: Between meaning and different methods. IntechOpen. https://doi.org/10.5772/intechopen.86587
Arham, Z., Kurniawan, K., Anhusadar, L., & Ismaun, I. (2021). Spectroscopic analysis of tempeh protein content during the production process. International Journal of Transdisciplinary Knowledge, 2(1), 51–62. https://doi.org/10.31332/ijtk.v2i1.17
Association of Official Agricultural Chemists (AOAC). (2005). Official methods of analysis of AOAC international (A. Incorporation (ed.)). Retrieved from https://scholar.google.co.id/scholar?cluster=9005071892713912602&hl=id&as_sdt=2005&sciodt=0,5
Beaubier, S., Albe-Slabi, S., Aymes, A., Bianeis, M., Galet, O., & Kapel, R. (2021). A rational approach for the production of highly soluble and functional sunflower protein hydrolysates. Foods, 10(3), 664. https://doi.org/10.3390/foods10030664
Cupp-Enyard, C. (2008). Sigma’s non-specific protease activity assay-casein as a substrate. Journal of Visualized Experiments, 19, e899. https://doi.org/10.3791/899
da Silva, P. P. M., Casemiro, R. C., Zillo, R. R., de Camargo, A. C., Prospero, E. T. P., & Spoto, M. H. F. (2014). Sensory descriptive quantitative analysis of unpasteurized and pasteurized jucara pulp (Euterpe edulis) during long-term storage. Food Science and Nutrition, 2(4), 321–331. https://doi.org/10.1002/fsn3.105
Djunaidi, S., Tirtaningtyas Gunawan-Puteri, M. D. P., Wijaya, C. H., & Prabawati, E. K. (2017). Physicochemical & microbial characterization of Overripe Tempeh. INSIST, 2(1), 48–51. https://doi.org/10.23960/ins.v2i1.33
Duong-Ly, K. C., & Gabelli, S. B. (2014). Salting out of proteins using ammonium sulfate precipitation. Methods in Enzymology, 541, 85–94. https://doi.org/10.1016/B978-0-12-420119-4.00007-0
Giguère, J. F., de Moura Piovesana, P., Proulx-Belhumeur, A., Doré, M., de Lemos Sampaio, K., & Gallani, M. C. (2016). Reliability of a simple method for determining salt taste detection and recognition thresholds. Chemical Senses, 41(3), 205–210. https://doi.org/10.1093/chemse/bjv080
Ginting, L., Wijanarka, & Kusdiyantini, E. (2020). Isolasi bakteri endofit tanaman pepaya (Carica papaya L.) dan uji aktivitas enzim amilase. Berkala Bioteknologi, 3(2), 1–7. Retrieved from https://ejournal2.undip.ac.id/index.php/bb/article/view/9654
Gunawan-Puteri, M. D. P. T., Hassanein, T. R., Prabawati, E. K., Wijaya, C. H., & Mutukumira, A. N. (2015). Sensory characteristics of seasoning powders from overripe tempeh, a solid state fermented soybean. Procedia Chemistry, 14, 263–269. https://doi.org/10.1016/j.proche.2015.03.037
Hassanein, T. R., Prabawati, E. K., & Gunawan-Puteri, D. M. T. P. (2015). Analysis of chemical and microbial changes during storage of overripe tempeh powder as seasoning material. International Journal of Science and Engineering (IJSE), 8(2), 131–134. https://doi.org/10.12777/ijse.8.2.131-134
Jeon, S. Y., Lee, Y. M., Kim, S. S., & Kim, K. O. (2020). Effect of added hydrolyzed vegetable proteins on consumers’ response for doenjang (Korean traditional fermented soybean paste) soup. Food Science and Biotechnology, 29(1), 45–53. https://doi.org/10.1007/s10068-019-00646-0
Kai, T., Sakamoto, Y., & Ishimoto, Y. (2015). Establishment of the proteolysis condition of raw material soybeans for tempeh production. Bulletin of Seinan Jo Gakuin University, 19, 97–108. Retrieved from https://seinan-jo.repo.nii.ac.jp
Khokhani, K., Ram, V., Bhatt, J., Khatri, T., & Joshi, H. (2012). Spectrophotometric and chromatographic analysis of amino acids present in leaves of Ailanthus excelsa. International Journal of ChemTech Research, 4(1), 389–393. Retrieved from https://www.cabdirect.org/cabdirect/abstract/20123082021
Laksono, A. S., Marniza, & Rosalina, Y. (2019). Karakteristik mutu tempe kedelai lokal varietas anjasmoro dengan variasi lama perebusan dan penggunaan jenis pengemas. Jurnal Agroindustri, 9(1), 8–18. https://doi.org/10.31186/j.agroind.9.1.8-18
Laskar, A., & Chatterjee, A. (2009). Protease–revisiting the types and potential. Journal of Biotechnology, 1(1), 55–61. Retrieved from https://scholar.google.com/scholar?hl=id&as_sdt=0%2C5&q=Protease+%E2%80%93+Revisiting+the+Types+and+Potential&btnG=
Lo, D., Romulo, A., Lin, J. Y., Wang, Y. T., Wijaya, C. H., & Wu, M. C. (2022). Effect of different fermentation conditions on antioxidant capacity and isoflavones content of soy tempeh. AIMS Agriculture and Food, 7(3), 567–579. https://doi.org/10.3934/agrfood.2022035
Machin, A. (2012). The potency of tempe hydrolysate as a flavor enhancer by utilization of the pineapple extract. Biosantifika, 4(2), 71–77. Retrieved from https://journal.unnes.ac.id/nju/index.php/biosaintifika/article/view/2275
Malle, D., Telussa, I., & Lasamahu, A. A. (2015). Isolasi dan karakterisasi papain dari buah pepaya (Carica papaya L.) jenis daun kipas. Indonesian Journal of Chemical Research, 2(2), 182–189. Retrieved from https://ojs3.unpatti.ac.id/index.php/ijcr/article/view/1951
Marković, S., Milošević, J., Đurić, M., Lolić, A., & Polović, N. (2021). One-step purification and freeze stability of papain at acidic pH values. Archives of Biological Sciences, 73(1), 57–64. https://doi.org/10.2298/ABS201217001M
Maulidiah, D. M., & Fibrianto, K. (2019). Profil sensori produk MSG PT. X dengan variasi media air mineral dan kaldu ayam. Jurnal Pangan dan Agroindustri, 7(2), 37–46. https://doi.org/10.21776/ub.jpa.2019.007.02.4
Mayasari, E., Priyono, S., Ulfa, M., & Saloko, S. (2018). Identifikasi asam amino glutamat pada bumbu instan daun san-sakng (Albertisia papuana Becc ). Seminar Nasional Penerapan Ilmu Pengetahuan dan Teknologi, pp. 83–88. Retrieved from https://www.researchgate.net/publication/332707210_Identifikasi_Asam_Amino_Glutamat_Pada_Bumbu_Instan_Daun_San-sakng_Albertisia_papuana_Becc
Muliadi, J., & Anugrahati, N. A. (2022). Utilization of mushroom as the main ingredient of vegan petis with different types of flour. GCISTEM Proceeding, 1, 132–142. https://doi.org/10.56573/gcistem.v1i.2
Muthmainna, M., Sabang, S. M., & Supriadi, S. (2017). Pengaruh waktu fermentasi terhadap kadar protein dari tempe biji buah lamtoro gung (Leucaena leucocephala). Jurnal Akademika Kimia, 5(1), 50–54. https://doi.org/10.22487/j24775185.2016.v5.i1.8001
Muyassaroh, Rini, D. K., & Minah, F. N. (2020). Penentuan kadar protein pada Spirulina platensis menggunakan metode lowry dan kjeldah. Jurnal Teknik Kimia, 15(1), 40–46. https://doi.org/10.33005/jurnal_tekkim.v15i1.2304
Muzdalifah, D., Athaillah, Z. A., Nugrahani, W., & Devi, A. F. (2017). Colour and pH changes of tempe during extended fermentation. AIP Conference Proceedings, 1803, 020036. https://doi.org/10.1063/1.4973163
Nelson, D. L., & Cox, M. M. (2017). Lehninger: Principles of Biochemistry, Volume Edition 7th. W. H. Freeman. Retrieved from https://scholar.google.com/scholar?hl=id&as_sdt=0%2C5&q=Lehninger+Principles+of+Biochemistry.+7th+Edition+nelson&btnG=
Nurrahman. (2015). Evaluasi komposisi zat gizi dan senyawa antioksidan kedelai hitam dan kedelai kuning. Jurnal Aplikasi Teknologi Pangan, 4(3), 89–93. https://doi.org/10.17728/jatp.2015.17
Pathare, P. B., Opara, U. L., & Al-Said, F. A. J. (2013). Colour measurement and analysis in fresh and processed foods: A review. Food and Bioprocess Technology, 6, 36–60. https://doi.org/10.1007/s11947-012-0867-9
Perdani, A. W., & Utama, Z. (2020). Korelasi kadar asam fitat dan protein terlarut tepung tempe kedelai lokal kuning (Glycine max) dan hitam (Glycine soja) selama fermentasi. Prosiding Pendidikan Teknik Boga Busana FT UNY, 15(1). Retrieved from https://journal.uny.ac.id/index.php/ptbb/article/view/36483/14864
Permanasari, A. R., Yulistiani, F., Purnama, R. W., Widjaja, T., & Gunawan, S. (2018). The effect of substrate and enzyme concentration on the glucose syrup production from red sorghum starch by enzymatic hydrolysis. IOP Conference Series: Earth and Environmental Science, 160, 012002. https://doi.org/10.1088/1755-1315/160/1/012002
Puteri, N. E., Astawan, M., Palupi, N. S., Wresdiyati, T., & Takagi, Y. (2018). Characterization of biochemical and functional properties of water-soluble tempe flour. Food Science and Technology (Brazil), 38, 147–153. https://doi.org/10.1590/fst.13017
Putri, K. M., Winarti, S., & Djajati, S. (2020). Physicochemical and organoleptic characteristics of seasoning from tempe hydrolysates using long treatment of fermentation and proteolytic enzyme proportion. Nusantara Science and Technology Proceedings, 76–85. https://doi.org/10.11594/nstp.2020.0511
Ratnayani, K., Juwarni, A. A. S., Laksmiwati, A. M., & Dewi, I. G. K. S. P. (2015). Uji aktivitas protease getah labu siam dan talas serta perbandingannya terhadap getah pepaya. Jurnal Kimia, 9(2), 147–152. Retrieved from https://repositori.unud.ac.id/protected/storage/upload/repositori/0a899c0fcac1b7da8b9a3e81113d95db.pdf
Rohmah, D. P. M., Hadi, S., & Baktir, A. (2019). Pemurnian parsial dan kritalisasi papain dari getah Carica papaya. Jurnal Kimia Riset, 4(2), 152–160. https://doi.org/10.20473/jkr.v4i2.16902
Saraswanti Indo Genetech. (2022). Amino Acids. PT Saraswanti Indo Genetech. Retrieved from https://siglaboratory.com/id/services/food-feed/amino-acids/
Shen, C. H. (2019). Diagnostic molecular biology. Academic Press. Retrieved from https://books.google.co.id/books?hl=id&lr=&id=KGGdBgAAQBAJ&oi=fnd&pg=PP1&dq=Diagnostic+Molecular+Biology+shen&ots=cUyBvi6c-r&sig=u9dPYXZqOckEMnOV0mXJZdCreCM&redir_esc=y#v=onepage&q=Diagnostic%20Molecular%20Biology%20shen&f=false
Sine, Y., & Soetarto, E. S. (2018). Isolasi dan identifikasi kapang rhizopus pada tempe gude (Cajanus cajan L.). Savana Cendana, 3(4), 67–68. https://doi.org/10.32938/sc.v3i04.487
Sitanggang, A. B., Firdausi, N. Z., & Budijanto, S. (2021). Antioxidant activity of a mixture between water-soluble tempeh extract and whey powder that has undergone a maillard reaction. Future of Food: Journal on Food, Agriculture and Society, 9(5), 1–12. https://doi.org/10.17170/kobra-202110144897
Sitanggang, A. B., Lesmana, M., & Budijanto, S. (2020). Membrane-based preparative methods and bioactivities mapping of tempe based peptides. Food Chemistry, 329, 127193. https://doi.org/10.1016/j.foodchem.2020.127193
Suparno, Kusumadati, W., Giyanto, & Winerungan, S. A. (2018). Kajian konsentrasi inokulum ragi terhadap sifat kimia dan bakteri asam laktat tempe kacang gude (Cajanus cajan L.). Jurnal AGRI PEAT, 19(1), 37–47. https://doi.org/10.36873/agp.v19i01.154
Taqwdasbriliani, E. B., Hutabarat, J., & Arini, E. (2013). Pengaruh kombinasi enzim papain dan enzim bromelin terhadap pemanfaatan pakan dan pertumbuhan ikan kerapu macan (Epinephelus fuscogutattus). Journal of Aquaculture Management and Technology, 2(3), 76–85. Retrieved from https://ejournal3.undip.ac.id/index.php/jamt/article/view/4795
Utami, R., Wijaya, C. H., & Lioe, H. N. (2016). Taste of water-soluble extracts obtained from over-fermented tempe. International Journal of Food Properties, 19(9), 2063–2073. https://doi.org/10.1080/10942912.2015.1104509
Wahyudi, A. (2018). Pengaruh variasi suhu ruang inkubasi terhadap waktu pertumbuhan Rhizopus oligosporus pada pembuatan tempe kedelai. Jurnal Agrium, 3(1), 37–38. https://doi.org/10.31851/redoks.v3i1.2790
Wicaksono, L. A., & Winarti, S. (2021). Karakteristik penyedap rasa alami dari biji bunga matahari dan kupang putih dengan hidrolisis enzimatis. AGRITEKNO: Jurnal Teknologi Pertanian, 10(1), 64–73. https://doi.org/10.30598/jagritekno.2021.10.1.64
Yazid, E. A., & Nuha, B. U. (2017). Kadar protein terlarut pada ampas kedelai dari hasil proses pembuatan tempe dengan penambahan ekstrak kasar papain (crude papain). Jurnal Ners Community, 8(1), 45–52. https://doi.org/10.55129/jnerscommunity.v8i1.285
Zhang, Y., Venkitasamy, C., Pan, Z., Liu, W., & Zhao, L. (2017). Novel umami ingredients: Umami peptides and their taste. Journal of Food Science, 82(1), 16–23. https://doi.org/10.1111/1750-3841.13576
Zusfahair, Ningsih, D. R., & Habibah, F. N. (2014). Karakterisasi papain dari daun pepaya (Carica papaya L.). Molekul, 9(1), 44–55 http://doi.org/10.20884/1.jm.2014.9.1.149
Refbacks
- There are currently no refbacks.
