Isolation of Hydrolytic Bacteria from Black Glutinous Rice Wash Water
DOI:
https://doi.org/10.25134/quagga.v18i2.522Keywords:
Bacillus tequilensis, Black glutinous rice wash water, Enzyme screening, Hydrolytic bacteria, Hydrolytic indexAbstract
Natural sources for hydrolytic enzymes produced by bacteria in demand in industry were increased. Hydrolytic bacteria can be found in agricultural and household waste, like black glutinous wash water. This study aims to identify and evaluate the presence and hydrolytic potential of bacteria in washing waste from fermented black glutinous rice. Fermented black glutinous rice washing water was used as the sample for bacterial isolation. The obtained isolates were screened for hydrolytic activity using modified Bushnell–Haas Agar supplemented with specific substrates to assess cellulolytic, amylolytic, and pectinolytic activities. Hydrolytic activity was expressed as a hydrolytic index to determine the most potent isolate, which was subsequently identified at the molecular level. A total of eight bacterial isolates were successfully obtained, among which one isolate, designated NKH4, showed the highest hydrolytic potential based on screening results. Molecular identification revealed that NKH4 is closely related to Bacillus tequilensis, with a similarity value of 97.96%. This species has previously been reported to produce various hydrolytic enzymes. The selected isolate NKH4 exhibited a qualitative hydrolytic index of 0.89, demonstrating its ability to hydrolyze starch, cellulose, and pectin. Therefore, NKH4, identified as Bacillus tequilensis, holds strong potential as a producer of hydrolytic enzymes and may serve as a promising candidate for further biotechnological applications.
References
Abba, N., Sung, C. T. B., Paing, T. N., & Zuan, A. T. K. (2021). Wastewater from washed rice water as plant nutrient source: current understanding and knowledge gaps. Pertanika Journal of Science and Technology, 29(3), 1347–1369. https://doi.org/10.47836/pjst.29.3.11
Adeoyo, O. R. (2025). Bioeconomic Perspectives of Bacterial and Fungal Hydrolytic Enzymes: A Review. International Journal of Scientific Research in Biological Sciences, 12(1), 61–70. https://doi.org/10.26438/ijsrbs.v12i1.670
Adnan, Wahyuni, S., & Khaeruni, A. R. (2017). PENGUJIAN SIFAT AMILOLITIK DAN PROTEOLITIK DARI ISOLAT BAKTERI ASAM LAKTAT (BAL) HASIL FERMENTASI AIR CUCIAN BERAS MERAH (Oryza nivara) KULTIVAR WAKAWONDU. J. Sains Dan Teknologi Pangan, 2(5), 759–769.
Alrumman, S., Mostafa, Y. S. M., Al-Qahtani, S., & Taha, T. H. T. (2018). Hydrolytic Enzyme Production by Thermophilic Bacteria Isolated from Saudi Hot Springs. Open Life Sciences, 13(1), 470–480. https://doi.org/10.1515/biol-2018-0056
Asril, M., Oktaviani, I., & Leksikowati, S. (2019). Isolasi Bakteri Indigineous dari Limbah Cair Tahu dalam Mendegradasi Protein dan Melarutkan Fosfat. Jurnal Teknologi Lingkungan, 20(1), 67. https://doi.org/10.29122/jtl.v20i1.3132
Bamigbade, A. T., Adewale, I. O., & Bakare, M. K. (2014). Characterization of a Novel α -amylase from Bacillus macquariensis Isolated from Cassava Peels Dump Site. Biocatalysis and Agricultural Biotechnology, 4(4), 84–92. https://doi.org/10.5923/j.ajb.20140404.04
Billa, A. S., & Rahmayanti, R. (2023). Potensi Air Cucian Beras Ketan Hitam (Oryza sativa var. glutinosa) sebagai Media Alternatif Pertumbuhan Candida albicans. Jurnal Jeumpa, 10(2), 259–268. https://doi.org/10.33059/jj.v10i2.8506
Chen, Y., Chen, H., Zhong, Q., Huan, Y., Weijun, Y., & Chen, W. (2021). Determination of Microbial Diversity and Community Composition in Unfermented and Fermented Washing Rice Water by High ‑ Throughput Sequencing. Current Microbiology, 78(5), 1730–1740. https://doi.org/10.1007/s00284-021-02400-4
Choi, J. M., Han, S. S., & Kim, H. S. (2015). Industrial applications of enzyme biocatalysis: Current status and future aspects. Biotechnology Advances, 33(7), 1443–1454. https://doi.org/10.1016/j.biotechadv.2015.02.014
Dar, M. A., Pawar, K. D., Rajput, B. P., Rahi, P., & Pandit, R. S. (2019). Purification of a cellulase from cellulolytic gut bacterium, Bacillus tequilensis G9 and its evaluation for valorization of agro-wastes into added value byproducts. Biocatalysis and Agricultural Biotechnology, 20(April), 101219. https://doi.org/10.1016/j.bcab.2019.101219
Dumitru, M., & Habeanu, M. (2021). Production and Evaluation of Extracellular Enzymes From Bacillus Licheniformis in Different Raw Materials Used in Animal Feed. LXIV(1), 129–136.
Elbeltagy, A., Nishioka, K., Suzuki, H., Sato, T., Sato, Y. I., Morisaki, H., Mitsui, H., & Minamisawa, K. (2000). Isolation and characterization of endophytic bacteria from wild and traditionally cultivated rice varieties. Soil Science and Plant Nutrition, 46(3), 617–629. https://doi.org/10.1080/00380768.2000.10409127
Fachrial, E., Putri, R. R. J. S., Lister, I. N. E., Anggraini, S., Harmileni, Nugroho, T. T., & Saryono. (2020). Molecular identification of cellulase and protease producing bacillus tequilensis utmsa14 isolated from the geothermal hot spring in lau sidebuk debuk, North Sumatra, Indonesia. Biodiversitas, 21(10), 4719–4725. https://doi.org/10.13057/biodiv/d211035
Fusvita, A., Idris, S. A., & Fitriani. (2018). IDENTIFIKASI BAKTERI ASAM LAKTAT ( BAL ) DAN KADAR ALKOHOL PADA PENDAHULUAN Salah satu makanan di Indonesia berbahan dasar beras ketan hitam adalah tape ketan hitam ( Fermentated Black Glutinous Rice ) yang mengandung antosianin , fenol dan aktivitas anti. Journal of Nursing & Health, 9(3), 385–389.
Ghosh, S., & Chatterjee, S. (2020). Isolation and Characterization of Bacillus tequilensis from Gut Content of Perionyx excavatus and Evaluation of its Starch Hydrolyzing Property. Bioscience Biotechnology Research Communications, 13(2), 670–675. https://doi.org/10.21786/bbrc/13.2/45
Hardoim, P. R., van Overbeek, L. S., Berg, G., Pirttilä, A. M., Compant, S., Campisano, A., Döring, M., & Sessitsch, A. (2015). The Hidden World within Plants: Ecological and Evolutionary Considerations for Defining Functioning of Microbial Endophytes. Microbiology and Molecular Biology Reviews, 79(3), 293–320. https://doi.org/10.1128/mmbr.00050-14
Kapil, S., Bhattu, M., & Kumar, T. (2021). Biomolecular Characterization of Bacillus tequilensis A1C1 Isolated from Soil and Chromatographic Analysis of D-serine in its Cellular Fraction. Research Square, 1–23.
Kim, Y.-K., Lee, S.-C., Cho, Y.-Y., Oh, H.-J., & Ko, Y. H. (2012). Isolation of Cellulolytic Bacillus subtilis Strains from Agricultural Environments . ISRN Microbiology, 2012, 1–9. https://doi.org/10.5402/2012/650563
Laca, A., Mousia, Z., Díaz, M., Webb, C., & Pandiella, S. S. (2006). Distribution of microbial contamination within cereal grains. Journal of Food Engineering, 72(4), 332–338. https://doi.org/10.1016/j.jfoodeng.2004.12.012
Mano, H., & Morisaki, H. (2008). Endophytic bacteria in the rice plant. Microbes and Environments, 23(2), 109–117. https://doi.org/10.1264/jsme2.23.109
Mazzucotelli, C. A., Ponce, A. G., Kotlar, C. E., & Moreira, M. del R. (2013). Isolation and characterization of bacterial strains with a hydrolytic profile with potential use in bioconversion of agroindustial by-products and waste. Food Science and Technology, 33(2), 295–303. https://doi.org/10.1590/S0101-20612013005000038
Mustofa, A., Anam, C., Praseptiangga, D., & Sutarno. (2024). A comparative study on physicochemical properties of rice and starch of white rice, black rice and black glutinous rice. Food Research, 8, 55–65. https://doi.org/10.26656/fr.2017.8(S2).566
Nazar, A. R. S., Salehi, N., Karimi, Y., Kasra Kermanshahi, R., Beheshti, M., Reza, A., & Nazari, S. (2018). Assessing the Biological Inhibitors Effect on Crude Oil Wax Appearance Temperature Reduction. Journal of Petroleum Science and Technology, 8(2), 70–85. https://doi.org/10.22078/jpst.2017.2442.1418
Ni’matuzahroh, Affandi, M., Ramadhan, R., Fatimah, Supriyanto, A., Salamun, Nurhariyati, T., Geraldi, A., Junairiah, Setiawan, J., Agustiana, B. N. D., Priyatama, A. H., Atmanindya, K., Alawiyah, D. D., Putra, E. P., Khiftiyah, A. M., Sari, S. K., & Clement, C. (2025). Diversity of Vachellia nilotica rhizosphere bacteria in the savanna of Baluran National Park, Indonesia and their potential for hydrolytic enzyme production. Biodiversitas, 26(4), 1754–1767. https://doi.org/10.13057/biodiv/d260425
Ni’matuzahroh, Affandi, M., Supriyanto, A., Rustantina, B., Jaiyah, L. A., Rahmawati, A., Nurhayati, H., Sari, S. K., Khiftiyah, A. M., & Huri, D. (2024). Biodiversity of hydrolytic enzymes-producing soil bacteria from a Durian Park, Jombang, Indonesia: Beneficial prospect for sustainable agriculture. Biodiversitas, 25(1), 392–403. https://doi.org/10.13057/biodiv/d250146
Nzilibili, S. M. M., Ekodiyanto, M. K. H., Hardjanto, P., & Yudianto, A. (2018). Concentration and Purity DNA Spectrophotometer: Sodium Monofluorophosphate forensic impended effect. Egyptian Journal of Forensic Sciences, 8(1). https://doi.org/10.1186/s41935-018-0065-7
Osman, M., & Abd El-Galeel, M. (2008). Physical, Chemical, Thermal and Technological Properties of Some Egyptian Rice Varieties. Journal of Food and Dairy Sciences, 33(8), 5893–5909. https://doi.org/10.21608/jfds.2008.124946
Prasetya, I. A. W., Suryani, E. M., & Ningrum, R. K. (2025). Isolation and Identification of Endophytic Cellulolytic Bacteria in Syzygium aqueum Fruit. Jurnal Biota, 11(2), 196–206. https://doi.org/10.19109/biota.v11i2.26424
Saihani, A., Hapizah, S., Agribisnis, P. S., Tinggi, S., & Pertanian, I. (2015). 1) & 2).
Salsabilla, P. S., Sidiq, Y., Tyastuti, E. M., & Rahayu, T. (2024). Selection of Proteolytic Bacteria from Bonoloyo Public Burial Place (PBP), Banjarsari, Surakarta, Central Java. Quagga: Jurnal Pendidikan Dan Biologi, 16(1), 43–50. https://doi.org/10.25134/quagga.v16i1.57
Samtiya, M., Aluko, R. E., & Dhewa, T. (2020). Plant food anti-nutritional factors and their reduction strategies: an overview. Food Production, Processing and Nutrition, 2(1), 1–14. https://doi.org/10.1186/s43014-020-0020-5
Saputri, R. D., Tyastuti, E. M., Sidiq, Y., & Rahayu, T. (2023). Public Cemetery’s Potency as The Source of Proteolytic Bacteria. Quagga: Jurnal Pendidikan Dan Biologi, 15(2), 201–206. https://doi.org/10.25134/quagga.v15i2.44
Shukla, E., D. Bendre, A., & M. Gaikwad, S. (2022). Hydrolases: The Most Diverse Class of Enzymes. https://doi.org/10.5772/intechopen.102350
Sulfianti, Risman, & Inang Saputri. (2021). Analisis Npk Pupuk Organik Cair Dari Berbagai Jenis Air Cucian Beras Dengan Metode Fermentasi Yang Berbeda. Jurnal Agrotech, 11(1), 36–42. https://doi.org/10.31970/agrotech.v11i1.62
Suryani, E. M., Jatmiko, Y. D., & Mustafa, I. (2023). Detection of Plantaricin-Encoding Gene and Its Partial Purification in Lactobacillus plantarum BP102. Jurnal Biodjati, 8(2), 233–247. https://doi.org/10.15575/biodjati.v8i2.27851
Tiwari, S., Prasad, V., & Lata, C. (2019). Bacillus: Plant growth promoting bacteria for sustainable agriculture and environment. In New and Future Developments in Microbial Biotechnology and Bioengineering: Microbial Biotechnology in Agro-environmental Sustainability. Elsevier B.V. https://doi.org/10.1016/B978-0-444-64191-5.00003-1
Urvoy, M., Lami, R., Dreanno, C., Delmas, D., L’Helguen, S., & Labry, C. (2021). Quorum Sensing Regulates the Hydrolytic Enzyme Production and Community Composition of Heterotrophic Bacteria in Coastal Waters. Frontiers in Microbiology, 12(December). https://doi.org/10.3389/fmicb.2021.780759
Wordu, G. O., Orisa, A. C., & Hamilton China, M. A. (2021). Nutrient and Anti-Nutrient Composition of Four Rice Varieties in Port Harcourt Metropolis. Asian Food Science Journal, 20(11), 90–100. https://doi.org/10.9734/afsj/2021/v20i1130379
Yang, D. C., Blair, K. M., & Salama, N. R. (2016). Staying in Shape: the Impact of Cell Shape on Bacterial Survival in Diverse Environments. Microbiology and Molecular Biology Reviews, 80(1), 187–203. https://doi.org/10.1128/mmbr.00031-15
Yunus, M. A., Raya, I., & Tuara, Z. I. (2019). Synthesis cellulose from rice husk. Indonesia Chimica Acta, 12(2), 79–83.
Zaki, D. N. H. (2018). Biodegradable plastic production by Bacillus spp. isolated from agricultural wastes and genetic analysis of PHA synthesis. Al-Mustansiriyah Journal of Science, 29(1), 67–74. https://doi.org/10.23851/mjs.v29i1.115
Zhang, G., Li, S., Xu, Y., Wang, J., Wang, F., Xin, Y., Shen, Z., Zhang, H., Ma, M., & Liu, H. (2019). Produ1. Zhang, G.; Li, S.; Xu, Y.; Wang, J.; Wang, F.; Xin, Y.; Shen, Z.; Zhang, H.; Ma, M.; Liu, H. Production of Alkaline Pectinase: A Case Study Investigating the Use of Tobacco Stalk with the Newly Isolated Strain Bacillus Tequilensis CAS-MEI-2-33. BM. BMC Biotechnology, 19(1), 1–11.
Zulkoni, A. (2014). Pemanfaatan Limbah Cucian Beras untuk Pembuatan Makanan Berserat Tinggi Menggunakan Bakteri Acetobacter xylinum. In Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati (pp. 83–91). https://doi.org/10.24002/biota.v18i2.391
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Indra Adi Wira Prasetya, Elsa Mega Suryani, Rizka Kusuma Ningrum
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
