ORIGINAL ARTICLE
Influence of Germination Time and UV Treatment on γ-Aminobutyric Acid Content and Microbial Dynamics of Germinated Red Rice in a Partially Circulated System
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1
Food Technology Study Program, Faculty of Engineering and Science, Universitas Pembangunan Nasional Veteran Jawa Timur, Surabaya 60294, Indonesia
2
Research Center for Food Technology and Processing, National Research and Innovation Agency (PRTPP-BRIN), Gunungkidul 55861, Indonesia
3
Department of Mechanical Engineering, Faculty of Engineering and Science, Universitas Pembangunan Nasional Veteran Jawa Timur, Surabaya 60294, Indonesia
4
Department of Informatics, Faculty of Computer Science, Universitas Pembangunan Nasional Veteran Jawa Timur, Surabaya 60294, Indonesia
Submission date: 2026-03-08
Acceptance date: 2026-07-27
Corresponding author
Hadi Munarko
Food Technology Study Program, Universitas Pembangunan Nasional Veteran Jawa Timur, JALAN RUNGKUT MADYA, GUNUNGANYAR, SURABAYA, 60294, Surabaya, Indonesia
KEYWORDS
TOPICS
ABSTRACT
Germination is a promising strategy to enhance bioactive compound content, particularly γ-aminobutyric acid (GABA) in red rice. However, conventional soaking germination methods are associated with high water consumption and microbial proliferation. This study evaluated the use of a partially circulated germinator with integrated microfiltration and ultraviolet (UV) treatment to process red rice for GABA accumulation while monitoring microbial dynamics. Red rice was germinated for 24, 48, and 72 h under UV and non-UV conditions, with water circulation applied at 90 min intervals for 5 min, and subsequently evaluated for pH, germination performance parameters, GABA content, and microbial quality. Microbial community diversity was characterized in one selected sample using next-generation sequencing (NGS). The results showed that the germinator effectively maintained water pH, mitigated drastic acidification commonly observed in static soaking systems. GABA content increased significantly with germination time, reaching maximum values at 48 h in both systems. Moreover, UV treatment resulted in higher GABA levels at 24 h. Total plate counts in circulation water exceeded 4 log CFU/mL, while reached 9.44 log CFU/g in dried germinated rice. UV treatment of the circulating water successfully reduced microbial load at 24 h germination time. Metagenomic profiling of freshly germinated red rice under non-UV treatment at 48 h revealed dominance of Phytobacter and Enterobacter, linked to seedling growth promotion, followed by Cronobacter spp., which presence suggested potential safety considerations. The findings demonstrate that partial circulation effectively enhances the functional value of red rice with reduced water consumption, offering a more environmentally sustainable alternative to conventional soaking methods.
ACKNOWLEDGEMENTS
The authors would like to acknowledge by the Ministry of Higher Education, Science, and Technology of Indonesia; Faculty of Engineering and Science, Universitas Pembangunan Nasional “Veteran” Jawa Timur; and the Research Center for Food Technology and Processing, BRIN for supporting this research.
FUNDING
This research was funded by the Ministry of Higher Education, Science, and Technology of Indonesia through the “Penelitian Fundamental Reguler/Regular Fundamental Research Grant Scheme” (Contract No. 098/C3/DT.05.00/PL/2025). This work also supported by Perjanjian Kerja Sama (PKS) / Cooperation Agreement between the Faculty of Engineering and Science, Universitas Pembangunan Nasional “Veteran” Jawa Timur, and the Research Center for Food Technology and Processing, BRIN, titled “Pengembangan Pangan Fungsional Berbasis Komoditas Lokal dan Kemasan Cerdas… / Development of Functional Foods from Local Commodities and Smart Packaging…”, under PKS Numbers 465/V/KS/10/2025 and 77/UN63.3/PKS/TU/2025.
CONFLICT OF INTEREST
The authors state no conflict of interests
ADDITIONAL INFORMATION
The authors declare they have not used Artificial Intelligence (AI) tools in the creation of this article. The AI, specifically ChatGPT (version GPT-4, OpenAI), was used to improve the readability and language of the article. The whole content has been reviewed and verified by the authors to ensure accuracy and integrity.
REFERENCES (52)
1.
Abdelfattah, A., Wisniewski, M., Schena, L., Tack, A.J.M. (2021). Experimental evidence of microbial inheritance in plants and transmission routes from seed to phyllosphere and root. Environmental Microbiology, 23(4), 2199–2214.
https://doi.org/10.1111/1462-2....
2.
Ansari, M.I., Jalil, S.U., Ansari, S.A., Hasanuzzaman, M. (2021). GABA shunt: a key-player in mitigation of ROS during stress. Plant Growth Regulation, 94(2), 131–149.
https://doi.org/10.1007/s10725....
3.
Baptista, E., Liberal, Â., Cardoso, R.V.C., Fernandes, Â., Dias, M.I., Pires, T.C.S.P., Calhelha, R.C., García, P.A., Ferreira, I.C.F.R., Barreira, J.C.M. (2024). Chemical and bioactive properties of red rice with potential pharmaceutical use. Molecules, 29(10), art. no. 2265.
https://doi.org/10.3390/molecu....
4.
Billington, C., Kingsbury, J.M., Rivas, L. (2022). Metagenomics approaches for improving food safety: A review. Journal of Food Protection, 85(3), 448–464.
https://doi.org/10.4315/JFP-21....
5.
Cebrián, G., Condón, S., Mañas, P. (2017). Physiology of the inactivation of vegetative bacteria by thermal treatments: mode of action, influence of environmental factors and inactivation kinetics. Foods, 6(12), art. no. 107.
https://doi.org/10.3390/foods6....
6.
Cechin, C. da F., Carvalho, G.G., Bastos, C.P., Kabuki, D.Y. (2023). Cronobacter spp. in foods of plant origin: occurrence, contamination routes, and pathogenic potential. Critical Reviews in Food Science and Nutrition, 63(33), 12398–12412.
https://doi.org/10.1080/104083....
7.
Chen, L., Bao, H., Yang, J., Huo, Y., Zhang, J., Fang, R., Zhang, L. (2024). Dynamics of rice seed-borne bacteria from acquisition to seedling colonization. Microbiome, 12(1), art. no. 253.
https://doi.org/10.1186/s40168....
8.
Chen, Z., Theppawong, A., Sangsawad, P., Fang, J., Ye, H., Deng, S., Yang, M., Gao, J., Kraithong, S. (2025). Bioactive compounds in colored rice: Exploring natural agents for cancer prevention in vitro and rodent model studies. Journal of Functional Foods, 129, art. no. 106875.
https://doi.org/10.1016/j.jff.....
9.
Chungcharoen, T., Sansiribhan, S., Munsin, R., Phetpan, K., Fonghiransiri, S., Limmun, W. (2024). The improvement of germination method for producing the germinated brown rice using a water spraying system with a revolved sieve. Engineering and Applied Science Research, 51(6), 739–746.
https://doi.org/10.14456/easr.....
10.
Cotter, P.D., Hill, C. (2003). Surviving the acid test: Responses of Gram-positive bacteria to low pH. Microbiology and Molecular Biology Reviews, 67(3), 429–453.
https://doi.org/10.1128/MMBR.6....
11.
Devi, P.A., Kalaiselvi, T. (2020). Evaluating the effect of Sphingobium yanoikuyae MH394206 and mixed consortia on growth of rice CO 51 in moisture deficit condition. Journal of Pharmacognosy and Phytochemistry, 9(6), 2016–2021.
https://doi.org/10.22271/phyto....
12.
Dutta, S., Choi, S.Y., Lee, Y.H. (2022). Temporal dynamics of endogenous bacterial composition in rice seeds during maturation and storage, and spatial dynamics of the bacteria during seedling growth. Frontiers in Microbiology, 13, art. no. 877781.
https://doi.org/10.3389/fmicb.....
13.
Eid, A.M., Fouda, A., Abdel‐Rahman, M.A., Salem, S.S., Elsaied, A., Oelmüller, R., Hijri, M., Bhowmik, A., Elkelish, A., El‐Din Hassan, S. (2021). Harnessing bacterial endophytes for promotion of plant growth and biotechnological applications: An overview. Plants, 10(5), art. no. 935.
https://doi.org/10.3390/plants....
14.
Gaveau, A., Coetsier, C., Roques, C., Bacchin, P., Dague, E., Causserand, C. (2017). Bacteria transfer by deformation through microfiltration membrane. Journal of Membrane Science, 523, 446–455.
https://doi.org/10.1016/j.mems....
15.
Jana, S.K., Bhattacharya, R., Mukherjee, S., Gupta, S., Hui, S.P., Chattopadhyay, A., Biswas, S.R., Mandal, S. (2025). Phytobacter sp. RSE02 is a rice seed endophytic plant probiotic bacterium with human probiotic features and cholesterol-lowering ability. Scientific Reports, 15(1), art. no. 27865.
https://doi.org/10.1038/s41598....
16.
Jiamyangyuen, S., Ooraikul, B. (2007). The physico-chemical, eating and sensorial properties of germinated brown rice. Journal of Food Agriculture & Environment, 6(2), 119–124.
17.
Kaga, H., Mano, H., Tanaka, F., Watanabe, A., Kaneko, S., Morisaki, H. (2009). Rice seeds as sources of endophytic bacteria. Microbes and Environments, 24(2), 154–162.
https://doi.org/10.1264/jsme2.....
18.
Kim, K.-S., Kim, B.-H., Kim, M.-J., Han, J.-K., Kum, J.-S., Lee, H.-Y. (2012). Quantitative microbiological profiles of brown rice and germinated brown rice. Food Science and Biotechnology, 21(6), 1785–1788.
https://doi.org/10.1007/s10068....
19.
Koutsoumanis, K.P., Lianou, A., Sofos, J.N. (2014). Food safety: Emerging pathogens. In N.K. Van Alfen (Ed.). Encyclopedia of Agriculture and Food Systems (Vol. 3), Academic Press, pp. 250–272.
https://doi.org/10.1016/B978-0....
20.
Kováříková, E., Ny, V., Šulc, M., Rysová, J., Pečenková, N., Houška, M. (2024). Promotional effects on naturally occurring lactic acid bacteria without impairing chickpea germination. Czech Journal of Food Sciences, 42(2), 85–92.
https://doi.org/10.17221/12/20....
21.
Kowalski, W. (2009). Chapter 2: UVGI Disinfection theory. In Ultraviolet Germicidal Irradiation Handbook. Springer Berlin Heidelberg, pp. 17-50.
https://doi.org/10.1007/978-3-....
22.
Kubota, H., Nakayama, T., Ariyoshi, T., Uehara, S., Uchitani, Y., Tsuchida, S., Nishiyama, H., Morioka, I., Koshinaga, T., Kusabuka, A., Nakatsubo, N., Yamagishi, T., Tabuchi, Y., Okuno, R., Kobayashi, K., Mitobe, M., Yokoyama, K., Shinkai, T., Suzuki, J., Sadamasu, K. (2023). Emergence of Phytobacter diazotrophicus carrying an IncA/C 2 plasmid harboring bla NDM-1 in Tokyo, Japan. mSphere, 8(4), art. no. e00147-23.
https://doi.org/10.1128/mspher....
23.
Kumar, N., Tripathi, K., Srivastava, Y. (2025). Effects of ultraviolet (UV) radiation on microbial growth: Mechanisms, responses, and applications. In K. Tripathi, Y. Srivastava, N. Kumar (Eds.), Biotechnology Lab Techniques: Culture Media, Microscopy, and Microbial Analysis, Deep Science Publishing, pp. 57–59.
https://doi.org/10.70593/978-9....
24.
Kurnianto, M.A., Adirama, S.I., Xu, W., Winarti, S., Rini, D.M. (2025). Enhancing the quality of traditional indonesian shrimp paste (terasi) through Tetragenococcus halophilus 54M106-3 inoculation: Physicochemical, sensory, and bioactivity insights. Foods, 14(14), art. no. 2419.
https://doi.org/10.3390/foods1....
25.
Liwinski, T., Lang, U.E., Brühl, A.B., Schneider, E. (2023). Exploring the therapeutic potential of gamma-aminobutyric acid in stress and depressive disorders through the gut–brain axis. Biomedicines, 11(12), art. no. 3128.
https://doi.org/10.3390/biomed....
26.
Maturin, L., Peeler, J.T. (2001). Chapter 3: Aerobic Plate Count. In Bacteriological Analytical Manual. U.S. Food & Drug Administration.
https://www.fda.gov/food/labor....
27.
Moore, M., Yang, T., Douraki, M.J., Rivard, C., Pliakoni, E., Nwadike, L., Bhullar, M. (2025). Effect of ultraviolet water treatment on survival and growth of Escherichia coli in recirculating hydroponic systems. Journal of Food Protection, 88(9), art. no. 100575.
https://doi.org/10.1016/j.jfp.....
28.
Munarko, H., Kurnianto, M.A., Jariyah, J., Arwani, A., Sari, R.M. (2025). Physicochemical properties, sensory profile, and emotional perception of unpolished organic rice. Polish Journal of Food and Nutrition Sciences, 75(2), 144–158.
https://doi.org/10.31883/pjfns....
29.
Munarko, H., Sitanggang, A.B., Kusnandar, F., Budijanto, S. (2021). Effect of different soaking and germination methods on bioactive compounds of germinated brown rice. International Journal of Food Science & Technology, 56(9), 4540–4548.
https://doi.org/10.1111/ijfs.1....
30.
Munarko, H., Sitanggang, A.B., Kusnandar, F., Budijanto, S. (2022). Germination of five Indonesian brown rice: evaluation of antioxidant, bioactive compounds, fatty acids and pasting properties. Food Science and Technology, 42, art. no. e19721.
https://doi.org/10.1590/fst.19....
31.
Nana, R., Maïga, Y., Ouédraogo, R.F., Kaboré, W.G.B., Badiel, B., Tamini, Z. (2019). Effect of water quality on the germination of okra (Abelmoschus esculentus) seeds. International Journal of Agronomy, 2019, art. no. 4938349.
https://doi.org/10.1155/2019/4....
32.
Nanfack, A.D., Nguefack, J., Musonerimana, S., La China, S., Giovanardi, D., Stefani, E. (2024). Exploiting the microbiome associated with normal and abnormal sprouting rice (Oryza sativa L.) seed phenotypes through a metabarcoding approach. Microbiological Research, 279, art. no. 127546.
https://doi.org/10.1016/j.micr....
33.
Ng, L.C., Sariah, M., Sariam, O., Radziah, O., Abidin, M.A.Z. (2012). Rice seed bacterization for promoting germination and seedling growth under aerobic cultivation system. Australian Journal of Crops Science, 6(1), 170-175.
34.
Ngalimat, M.S., Mohd Hata, E., Zulperi, D., Ismail, S.I., Ismail, M.R., Mohd Zainudin, N.A.I., Saidi, N.B., Yusof, M.T. (2021). Plant growth-promoting bacteria as an emerging tool to manage bacterial rice pathogens. Microorganisms, 9(4), art. no. 682.
https://doi.org/10.3390/microo....
35.
Nooshabadi, R., Hashemi, S.M. (2024). The effect of ultraviolet radiation on pathogenic microorganisms in milk, traditional fruit juice, and drinking water. Food Hygiene, 14(54), fa21-fa33.
https://doi.org/10.71876/jfh.2....
36.
Osaili, T.M., Shaker, R.R., Al-Haddaq, M.S., Al-Nabulsi, A.A., Holley, R.A. (2009). Heat resistance of Cronobacter species (Enterobacter sakazakii) in milk and special feeding formula. Journal of Applied Microbiology, 107(3), 928–935.
https://doi.org/10.1111/j.1365....
37.
Pearce, L.E., Smythe, B.W., Crawford, R.A., Oakley, E., Hathaway, S.C., Shepherd, J.M. (2012). Pasteurization of milk: The heat inactivation kinetics of milk-borne dairy pathogens under commercial-type conditions of turbulent flow. Journal of Dairy Science, 95(1), 20–35.
https://doi.org/10.3168/jds.20....
38.
Quibod, I.L., Grande, G., Oreiro, E.G., Borja, F.N., Dossa, G.S., Mauleon, R., Cruz, C.V., Oliva, R. (2015). Rice-infecting pseudomonas genomes are highly accessorized and harbor multiple putative virulence mechanisms to cause sheath brown rot. PLoS One, 10(9), art. no. e0139256.
https://doi.org/10.1371/journa....
39.
Ramírez-Olvera, S.M., Trejo-Téllez, L.I., García-Morales, S., Pérez-Sato, J.A., & Gómez-Merino, F.C. (2018). Cerium enhances germination and shoot growth, and alters mineral nutrient concentration in rice. PLoS One, 13(3), art. no. e0194691.
https://doi.org/10.1371/journa....
40.
Rodrigues, L.A., Cañizares, L. da C.C., Meza, S.L.R., Peres, B.B., Jappe, S.N., Timm, N. da S., Oliveira, M. de, Coradi, P.C. (2024). A review of the influence of genotype, environment, and food processing on the bioactive compound profile of red rice (Oryza sativa L.). Agronomy, 14(3), art. no. 616.
https://doi.org/10.3390/agrono....
41.
Rohman, A., Gandjar, I.G. (2007). Chromatography Methods for Food Analysis. Pustaka Pelajar, Yogyakarta, pp. 48-54.
43.
Scarlett, K., Collins, D., Tesoriero, L., Jewell, L., van Ogtrop, F., Daniel, R. (2016). Efficacy of chlorine, chlorine dioxide and ultraviolet radiation as disinfectants against plant pathogens in irrigation water. European Journal of Plant Pathology, 145(1), 27–38.
https://doi.org/10.1007/s10658....
44.
Shelp, B.J., Bozzo, G.G., Trobacher, C.P., Zarei, A., Deyman, K.L., Brikis, C.J. (2012). Hypothesis/review: Contribution of putrescine to 4-aminobutyrate (GABA) production in response to abiotic stress. Plant Science, 193–194, 130–135.
https://doi.org/10.1016/j.plan....
45.
Sitanggang, A.B., Joshua, M., Munarko, H., Kusnandar, F., Budijanto, S. (2021). Increased γ-aminobutyric acid content of germinated brown rice produced in membrane reactor. Food Technology and Biotechnology, 59(3), 295–305.
https://doi.org/10.17113/ftb.5....
46.
Tariq, H., Subramanian, S., Geitmann, A., Smith, D.L. (2025). Bacillus and Paenibacillus as plant growth-promoting bacteria in soybean and cannabis. Frontiers in Plant Science, 16, art. no. 1529859.
https://doi.org/10.3389/fpls.2....
47.
Wang, Y., Li, M., Xu, F., Chai, L., Bao, J., Shen, S. (2016). Variation in polyphenols, tocols, γ‐aminobutyric acid, and antioxidant properties in whole grain rice (Oryza sativa L.) as affected by different germination time. Cereal Chemistry, 93(3), 268–274.
https://doi.org/10.1094/CCHEM-....
48.
Yan, L., Zheng, H., Liu, W., Liu, C., Jin, T., Liu, S., Zheng, L. (2021). UV-C treatment enhances organic acids and GABA accumulation in tomato fruits during storage. Food Chemistry, 338, art. no. 128126.
https://doi.org/10.1016/j.food....
49.
Yang, G., Xu, J., Xu, Y., Li, R., Wang, S. (2023). Analysis of dynamics and diversity of microbial community during production of germinated brown rice. Foods, 12(4), art. no. 755.
https://doi.org/10.3390/foods1....
50.
Zhang, C., Xia, X., Li, B., Hung, Y.-C. (2018). Disinfection efficacy of electrolyzed oxidizing water on brown rice soaking and germination. Food Control, 89, 38–45.
https://doi.org/10.1016/j.food....
51.
Zhu, J., Li, C., Sun, L., Cheng, Y., Hou, J., Fan, Y., Ge, Y. (2022). Application of γ-aminobutyric acid induces disease resistance in apples through regulation of polyamine metabolism, GABA shunt and reactive oxygen species metabolism. Scientia Horticulturae, 291, art. no. 110588.
https://doi.org/10.1016/j.scie....
52.
Zhu, T., Ruan, S., Zhang, J., Zhou, B., Fang, L., Song, K., Tu, S., Tu, K. (2023). γ-Aminobutyric acid (GABA) mediated stilbene biosynthesis and alleviated oxidative stress induced by UV-C radiation during peanuts (Arachis hypogaea L.) germination. Acta Physiologiae Plantarum, 45(8), art. no. 102.
https://doi.org/10.1007/s11738....