Investigation of the Physicochemical Properties and its Correlation during Koji-Moromi Fermentation Stage of Production Soy Sauce Naturally Brewed in Central Java, Indonesia

Bhakti Etza Setiani (1), Yunianta (2), Elok Zubaidah (3), Agustin Krisna Wardani (4)
(1) Doctoral Program of Food Science Department, Faculty of Agriculture Technology, Brawijaya University, Malang, 65145, Indonesia
(2) Food Science Department, Faculty of Agriculture Technology, Brawijaya University, Malang, 65145, Indonesia
(3) Food Science Department, Faculty of Agriculture Technology, Brawijaya University, Malang, 65145, Indonesia
(4) Food Science Department, Faculty of Agriculture Technology, Brawijaya University, Malang, 65145, Indonesia
Fulltext View | Download
How to cite (IJASEIT) :
Setiani, Bhakti Etza, et al. “Investigation of the Physicochemical Properties and Its Correlation During Koji-Moromi Fermentation Stage of Production Soy Sauce Naturally Brewed in Central Java, Indonesia”. International Journal on Advanced Science, Engineering and Information Technology, vol. 14, no. 2, Apr. 2024, pp. 768-76, doi:10.18517/ijaseit.14.2.19559.
Soy sauce, a widely consumed condiment, undergoes intricate physicochemical transformations during its natural fermentation process. This research delved into the analysis of physical and chemical properties in naturally brewed soy sauce, aiming to elucidate the impact of fermentation stages (Koii, earlier moromi, final moromi, soy sauce product) and sample variations on these properties. Parameters such as pH, total acid, total soluble solids, and water activity (aw) were assessed. Additionally, chemical attributes were analyzed, including total nitrogen, crude protein, formol nitrogen, and sodium chloride (NaCl) content. The correlation between physical and chemical properties during the fermentation stages was determined using the principal component analysis (PCA) method. The fermentation process significantly affected soy sauce's physical and chemical properties. Key factors include total dissolved solids, water activity, total acids, salt, total nitrogen, crude protein, and formol nitrogen, which display distinct patterns. These fluctuations were primarily driven by microbial activity and enzymatic hydrolysis, and total nitrogen, crude protein, and formol nitrogen play pivotal roles in characterizing soy sauce quality. Remarkably, total acid content and pH values exhibited significant variations that deviate from expected correlations due to weak acids. Samples C and D were deemed the most suitable out of the five tested, aligning better with quality standards for soy sauce. This study sheds light on the complex physicochemical changes during soy sauce fermentation, emphasizing the impact of raw materials and microbial processes on its properties.

N. Aryana, D. P. Ramadhaningtyas, D. Styarini, and Y. Aristiawan, “First Indonesian proficiency testing using reference value from isotope dilution mass spectrometry method for benzoic acid, methyl paraben, and n-butyl paraben in sweet soy sauce,” Accreditation Qual. Assur., vol. 24, no. 6, pp. 427–435, Dec. 2019, doi: 10.1007/s00769-019-01398-4.

B. S. Luh, “Industrial production of soy sauce,” J. Ind. Microbiol., vol. 14, no. 6, pp. 467–471, Jun. 1995, doi: 10.1007/BF01573959.

C. Diez-Simon, C. Eichelsheim, R. Mumm, and R. D. Hall, “Chemical and Sensory Characteristics of Soy Sauce: A Review,” J. Agric. Food Chem., vol. 68, no. 42, pp. 11612–11630, Oct. 2020, doi:10.1021/acs.jafc.0c04274.

P. V. P. Devanthi and K. Gkatzionis, “Soy sauce fermentation: Microorganisms, aroma formation, and process modification,” Food Res. Int., vol. 120, pp. 364–374, Jun. 2019, doi:10.1016/j.foodres.2019.03.010.

L. Zhang, S. Xiong, T. Du, M. Xiao, Z. Peng, M. Xie, Q. Guan, and T. Xiong, “Effects of microbial succession on the dynamics of flavors metabolites and physicochemical properties during soy sauce koji making,” Food Biosci., vol. 53, p. 102636, April. 2023, doi:10.1016/j.fbio.2023.102636.

S. Sassi, W. A. A. Q. I. Wan‐Mohtar, N. S. Jamaludin, and Z. Ilham, “Recent progress and advances in soy sauce production technologies: A review,” J. Food Process. Preserv., vol. 45, no. 10, Oct. 2021, doi: 10.1111/jfpp.15799.

R. Y. Zhou, J.-Y. Chua, and S.-Q. Liu, “Exploring the feasibility of biotransforming salted soy whey into a soy sauce-like condiment using wine yeast Torulaspora delbrueckii and soy sauce yeasts Zygosaccharomyces rouxii and Candida versatilis as single starter cultures,” Food Res. Int., vol. 156, p. 111350, Jun. 2022, doi:10.1016/j.foodres.2022.111350.

C. Chen, S. Hou, C. Wu, Y. Cao, X. Tong, and Y. Chen, “Improving protein utilization and fermentation quality of soy sauce by adding protease,” J. Food Compos. Anal., vol. 121, p. 105399, Aug. 2023, doi: 10.1016/j.jfca.2023.105399.

H. Guo, Z. Li, J. Huang, R. Zhou, C. Wu, and Y. Jin, “Microfiltration of soy sauce: Efficiency, resistance and fouling mechanism at different operating stages,” Sep. Purif. Technol., vol. 240, p. 116656, Jun. 2020, doi: 10.1016/j.seppur.2020.116656.

G. J. Kaur, V. Orsat, and A. Singh, “An overview of different homogenizers, their working mechanisms and impact on processing of fruits and vegetables,” Crit. Rev. Food Sci. Nutr., vol. 63, no. 14, pp. 2004–2017, May 2023, doi: 10.1080/10408398.2021.1969890.

K. Ito and A. Matsuyama, “Koji Molds for Japanese Soy Sauce Brewing: Characteristics and Key Enzymes,” J. Fungi, vol. 7, no. 8, p. 658, Aug. 2021, doi: 10.3390/jof7080658.

L. Ruan, Y. Ju, C. Zhan, and L. Hou, “Improved umami flavor of soy sauce by adding enzymatic hydrolysate of low-value fish in the natural brewing process,” LWT, vol. 155, p. 112911, Feb. 2022, doi:10.1016/j.lwt.2021.112911.

D. A. Anagnostopoulos, F. F. Parlapani, E. Tsara, M. G. Eirinaki, D. Kokioumi, E. Ampatzidou, and I. S. Boziaris, “Effect of Physicochemical Characteristics and Storage Atmosphere on Microbiological Stability and Shelf-Life of Minimally Processed European Sea Bass (Dicentrarchus labrax) Fillets,” Foods, vol. 12, no. 6, p. 1145, Mar. 2023, doi: 10.3390/foods12061145.

G. Tan, Y. Wang, M. Hu, X. Li, X. Li, Z. Pan, M. Li, L. Li, and Z. Zheng, “Comparative evaluation of the microbial diversity and metabolite profiles of Japanese-style and Cantonese-style soy sauce fermentation,” Front. Microbiol., vol. 13, p. 976206, Aug. 2022, doi:10.3389/fmicb.2022.976206.

(BSN) Badan Standarisasi Nasional, “SNI 3543.1-2013. Kecap kedelai – Bagian 1: Manis.” Badan Standarisasi Nasional, Jakarta, 2013.

A. A. de Brito, F. campos, A. d. R. Nascimento, C. Damiani, F. A. da Silva, G. H. de A. Teixeira, and L. C. C. Junior, “Non-destructive determination of color, titratable acidity, and dry matter in intact tomatoes using a portable Vis-NIR spectrometer,” J. Food Compos. Anal., vol. 107, p. 104288, Apr. 2022, doi:10.1016/j.jfca.2021.104288.

M. Sezey and P. Adun, “Validation of Mohr Titration Method to Determine Salt in Olive and Olive Brine,” J. Turk. Chem. Soc. Sect. Chem., vol. 6, no. 3, pp. 329–334, Oct. 2019, doi:10.18596/jotcsa.496563.

N. J. Thiex, H. Manson, S. Andersson, and J.-A. Persson, “Determination of crude protein in animal feed, forage, grain, and oilseeds by using block digestion with a copper catalyst and steam distillation into boric acid: collaborative study,” J. AOAC Int., vol. 85, no. 2, pp. 309–317, 2002.

Badan Standarisasi Nasional (BSN), “SNI 01-2354.4-2006. Cara uji kimia - Bagian 4: Penentuan kadar protein dengan metode total nitrogen pada produk perikanan.” (BSN) Badan Standarisasi Nasional, Jakarta, 2006.

H. Sasaki, D. Pham Thi Ngoc, M. Nishikawa, and M. Kanauchi, “Lipopolysaccharide neutralizing protein in Miso , Japanese fermented soybean paste,” J. Food Sci., vol. 85, no. 8, pp. 2498–2505, Aug. 2020, doi: 10.1111/1750-3841.15315.

J. A. Tantray, S. Mansoor, R. F. C. Wani, and N. U. Nissa, “pH meter,” in Basic Life Science Methods, Elsevier, 2023, pp. 9–10. doi:10.1016/B978-0-443-19174-9.00003-9.

A. Blanco and G. Blanco, “Water,” in Medical Biochemistry, Elsevier, 2017, pp. 5–19. doi: 10.1016/B978-0-12-803550-4.00002-1.

X. Jiang, D. Peng, W. Zhang, M. Duan, Z. Ruan, S. Huang, S. Zhou, and Q. Fang, “Effect of aroma-producing yeasts in high-salt liquid-state fermentation soy sauce and the biosynthesis pathways of the dominant esters,” Food Chem., vol. 344, p. 128681, May 2021, doi:10.1016/j.foodchem.2020.128681.

R. Liang, J. Huang, X. Wu, J. Fan, Y. Xu, C. Wu, Y. Jin, and R. Zhou, “Effect of raw material and starters on the metabolite constituents and microbial community diversity of fermented soy sauce,” J. Sci. Food Agric., vol. 99, no. 13, pp. 5687–5695, Oct. 2019, doi:10.1002/jsfa.9830.

J. Li, B. Wang, J. Chen, G. Du, and F. Fang, “Heme Dependent Catalase Conditionally Contributes to Oxygen Tolerance of Tetragenococcus halophilus Strains Isolated from Soy Sauce Moromi,” Appl. Sci., vol. 12, no. 16, p. 8039, Aug. 2022, doi:10.3390/app12168039.

Q. Wang, R. Cui, X. Liu, X. Zheng, Y. Yao, and G. Zhao, “Examining the impact of Tetragenococcus halophilus , Zygosaccharomyces rouxii , and Starmerella etchellsii on the quality of soy sauce: a comprehensive review of microbial population dynamics in fermentation,” Crit. Rev. Food Sci. Nutr., pp. 1–12, Jul. 2023, doi:10.1080/10408398.2023.2230285.

J. G. Allwood, L. T. Wakeling, and D. C. Bean, “Fermentation and the microbial community of Japanese koji and miso : A review,” J. Food Sci., vol. 86, no. 6, pp. 2194–2207, Jun. 2021, doi: 10.1111/1750-3841.15773.

P. V. P. Devanthi, R. Linforth, H. El Kadri, and K. Gkatzionis, “Water-in-oil-in-water double emulsion for the delivery of starter cultures in reduced-salt moromi fermentation of soy sauce,” Food Chem., vol. 257, pp. 243–251, Aug. 2018, doi: 10.1016/j.foodchem.2018.03.022.

W. Zhang, Z. Xiao, Z. Gu, X. Deng, J. Liu, X. Luo, C. Song, and X. Jiang, "Fermentation-promoting effect of three salt-tolerant Staphylococcus and their co-fermentation flavor characteristics with Zygosaccharomyces rouxii in soy sauce brewing,” Food Chem., vol. 432, p. 137245, Jan. 2024, doi: 10.1016/j.foodchem.2023.137245.

J. Li, C. Sun, Z. Shen, Y. Tian, F. Mo, B. Wang, B. Liu, and C. Wang, “Untargeted metabolomic profiling of Aspergillus sojae 3.495 and Aspergillus oryzae 3.042 fermented soy sauce koji and effect on moromi fermentation flavor,” LWT, vol. 184, p. 115027, Jul. 2023, doi:10.1016/j.lwt.2023.115027.

R. Y. Zhou, X. Huang, Z. Liu, J.-Y. Chua, and S.-Q. Liu, “Evaluating the effect of lactic acid bacterial fermentation on salted soy whey for development of a potential novel soy sauce-like condiment,” Curr. Res. Food Sci., vol. 5, pp. 1826–1836, 2022, doi:10.1016/j.crfs.2022.10.004.

D. Ac and P. Kn, “Effect of Storage on Physicochemical Properties of Spiced Fish Sauce,” J. Nutr. Food Sci., vol. 6, no. 4, 2016, doi:10.4172/2155-9600.1000520.

F. N. Khasanah, D. F. Rosida, and R. A. Anggreini, “Physical Characteristics of Golden Apple Snail (Pomacea Canaliculata) Sauce using Bromelain and Calotropin Enzym,” Int. J. Eco-Innov. Sci. Eng., vol. 3, no. 02, pp. 45–48, Nov. 2022, doi: 10.33005/ijeise.v3i02.69.

A. Amar, S. Makosim, S. Sukotjo, N. Ahadiyanti, and E. Weisman, “Growth dynamics of mold-yeast and bacteria during the production process of saga tauco [Adenanthera pavonina],” IOP Conf. Ser. Earth Environ. Sci., vol. 741, no. 1, p. 012019, May 2021, doi:10.1088/1755-1315/741/1/012019.

Y. Jo, W. Bang, and M. K. Kim, “Changes of Physiochemical and Enzymatic Activities of doenjang Prepared with Different Amount of Rice koji during 30 Days of Fermentation,” Foods, vol. 10, no. 2, p. 372, Feb. 2021, doi: 10.3390/foods10020372.

Q.-Q. Lv, J.-J. Cao, R. Liu, and H.-Q. Chen, “Structural characterization, α-amylase and α-glucosidase inhibitory activities of polysaccharides from wheat bran,” Food Chem., vol. 341, p. 128218, Mar. 2021, doi: 10.1016/j.foodchem.2020.128218.

J. Li, H. Huang, W. Feng, R. Guan, L. Zhou, H. Cheng, and X. Ye, “Dynamic Changes in Biogenic Amine Content in the Traditional Brewing Process of Soy Sauce,” J. Food Prot., vol. 82, no. 9, pp. 1539–1545, Sep. 2019, doi: 10.4315/0362-028X.JFP-19-035.

Y. Tian, Y. Feng, M. Zhao, and G. Su, “Comparison and application of the extraction method for the determination of enzymatic profiles in matured soybean koji,” Food Biosci., vol. 49, p. 101875, Oct. 2022, doi: 10.1016/j.fbio.2022.101875.

L. Zhu, S. He, Y. Lu, J. Gan, N. Tao, X. Wang, and Z. Jiang, Y. Hong, and C. Xu, “Metabolomics mechanism of traditional soy sauce associated with fermentation time,” Food Sci. Hum. Wellness, vol. 11, no. 2, pp. 297–304, Mar. 2022, doi: 10.1016/j.fshw.2021.11.023.

J. S. Abdurrauf and B. Aceh, “Optimising Protein and Total Dissolved Solid to Synthesize Soy Sauce From Soybean Residue Using Box-Behnken Design,” vol. 14, 2019.

H. L. Zahidah and D. Lo, “The physicochemical properties of soy sauce made from tempeh,” IOP Conf. Ser. Earth Environ. Sci., vol. 1115, no. 1, p. 012094, Dec. 2022, doi: 10.1088/1755-1315/1115/1/012094.

M. S. Tapia, S. M. Alzamora, and J. Chirife, “Effects of Water Activity (aw) on Microbial Stability as a Hurdle in Food Preservation,” in Water Activity in Foods, G. V. Barbosa‐Cánovas, A. J. Fontana, S. J. Schmidt, and T. P. Labuza, Eds., 1st ed.Wiley, 2020, pp. 323–355. doi:10.1002/9781118765982.ch14.

Y. Wang, C. Li, L. Li, X. Yang, Y. Wu, Y. Zhao, and Y. Wei, “Effect of Bacterial Community and Free Amino Acids on the Content of Biogenic Amines During Fermentation of Yu-lu, a Chinese Fermented Fish Sauce,” J. Aquat. Food Prod. Technol., vol. 27, no. 4, pp. 496–507, Apr. 2018, doi: 10.1080/10498850.2018.1450573.

R. P. U. Dwipa, T. Maneeboon, C. Chuaysrinule, and W. Mahakarnchanakul, “Interactive Inhibition of Aflatoxigenic Aspergillus flavus and Ochratoxigenic Aspergillus carbonarius by Aspergillus oryzae under Fluctuating Temperatures,” Microbiol. Res., vol. 13, no. 1, pp. 114–127, Feb. 2022, doi:10.3390/microbiolres13010009.

A. Brambilla and A. Sangiorgio, “Mould growth in energy efficient buildings: Causes, health implications and strategies to mitigate the risk,” Renew. Sustain. Energy Rev., vol. 132, p. 110093, Oct. 2020, doi: 10.1016/j.rser.2020.110093.

Y. Ma, J. yi, J. Bi, X. Wu, X. Li, J. Li, and Y. Zhao, “Understanding of osmotic dehydration on mass transfer and physical properties of freeze‐dried apple slices: A comparative study of five saccharides osmotic agents,” J. Food Process. Preserv., vol. 46, no. 3, Mar. 2022, doi: 10.1111/jfpp.16328.

N. Fitri, S. X. Y. Chan, N. H. C. Lah, F. A. Jam, N. M. Misnan, N. Kamal, M. N. Sarian, M. A. M. Lazaldin, C. F. Low, H. S. Hamezah, E. R. Rohani, A. Mediani, and F. Abah, “A Comprehensive Review on the Processing of Dried Fish and the Associated Chemical and Nutritional Changes,” Foods, vol. 11, no. 19, p. 2938, Sep. 2022, doi:10.3390/foods11192938.

N. K. K. S. Dewi, I Wayan Sudiarta, and I Nyoman Rudianta, “Substitution of Corn Flour and Additional Palm Sugar to Cookies Characteristics,” SEAS Sustain. Environ. Agric. Sci., vol. 6, no. 1, pp. 42–52, May 2022, doi: 10.22225/seas.6.1.4965.42-52.

S. Seleshe and S. N. Kang, “Effect of Different Pediococcus pentosaceus and Lactobacillus plantarum Strains on Quality Characteristics of Dry Fermented Sausage after Completion of Ripening Period,” Food Sci. Anim. Resour., vol. 41, no. 4, pp. 636–649, Jul. 2021, doi: 10.5851/kosfa.2021.e21.

X. Kuang, H. Su, W. Li, L. Lin, W. Lin, and L. Luo, “Effects of microbial community structure and its co-occurrence on the dynamic changes of physicochemical properties and free amino acids in the Cantonese soy sauce fermentation process,” Food Res. Int., vol. 156, p. 111347, Jun. 2022, doi: 10.1016/j.foodres.2022.111347.

X. Zhao, Y. Liu, L. Shu, and Y. He, “Study on metabolites of Bacillus producing soy sauce‐like aroma in Jiang‐flavor Chinese spirits,” Food Sci. Nutr., vol. 8, no. 1, pp. 97–103, Jan. 2020, doi: 10.1002/fsn3.1266.

D. Drulyte and V. Orlien, “The Effect of Processing on Digestion of Legume Proteins,” Foods, vol. 8, no. 6, p. 224, Jun. 2019, doi:10.3390/foods8060224.

F. Urbat, P. Müller, A. Hildebrand, D. Wefers, and M. Bunzel, “Comparison and Optimization of Different Protein Nitrogen Quantitation and Residual Protein Characterization Methods in Dietary Fiber Preparations,” Front. Nutr., vol. 6, p. 127, Aug. 2019, doi: 10.3389/fnut.2019.00127.

Y. Feng, Y. Cai, G. Su, H. Zhao, C. Wang, and M. Zhao, “Evaluation of aroma differences between high-salt liquid-state fermentation and low-salt solid-state fermentation soy sauces from China,” Food Chem., vol. 145, pp. 126–134, Feb. 2014, doi:10.1016/j.foodchem.2013.07.072.

X. Gao, E. Liu, J. Zhang, M. Yang, S. Chen, Z. Liu, H. Ma, and F. Hu, “Effects of sonication during moromi fermentation on antioxidant activities of compounds in raw soy sauce,” LWT, vol. 116, p. 108605, Dec. 2019, doi: 10.1016/j.lwt.2019.108605.

X. Gao, J. Zhang, E. Liu, M. Yang, S. Chen, F. Hu, H. Ma, Z. Liu, and X. Yu, “Enhancing the taste of raw soy sauce using low intensity ultrasound treatment during moromi fermentation,” Food Chem., vol. 298, p. 124928, Nov. 2019, doi: 10.1016/j.foodchem.2019.05.202.

J.-Y. Ciou, L.-S. Hsieh, T.-T. Lee, and C.-W. Hsieh, “Enhancement of Agricultural Processed By-Products: Qualities Analysis of Fermentation Method in Gradient Salt Adding Treatment of Tuna Cooking Juice with Black Bean Koji Added,” Foods, vol. 9, no. 3, p. 320, Mar. 2020, doi: 10.3390/foods9030320.

Y. Funatsu, K. Azekawa, T. Ono, H. Hayasaka, W. Matsuda, N. Maeda, A. Tanaka and S. Yoshikawa, “Effect of soy sauce yeast inoculation and ureter removal on the quality characteristics of meat sauce prepared from pig kidneys,” Food Sci. Technol. Res., vol. 27, no. 2, pp. 259–268, 2021, doi: 10.3136/fstr.27.259.

B. Auttanak, P. Chomnawang, K. Huaisan, K. Sootsuwan, and C. Chomnawang, “Influences of salt and koji contents on chemical properties of fish sauce from tilapia frames,” Aquac. Fish., vol. 7, no. 1, pp. 67–73, Jan. 2022, doi: 10.1016/j.aaf.2020.11.005.

Z. Hao, L. Liang, D. Pu, and Y. Zhang, “Analysis of Sodium Content in 4082 Kinds of Commercial Foods in China,” Nutrients, vol. 14, no. 14, p. 2908, Jul. 2022, doi: 10.3390/nu14142908.

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with this journal agree to the following terms:

    1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
    2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
    3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).