EFFECTS OF PURE AND COMMERCIAL PROBIOTIC CULTURES ON GAMMA-AMINOBUTYRIC ACID (GABA) FORMATION AND QUALITY ATTRIBUTES OF THAI FERMENTED PORK SAUSAGE (NHAM)
Abstract
This study investigated the effects of pure and commercial probiotic starter cultures on gamma-aminobutyric acid (GABA) formation and key quality attributes of Thai fermented pork sausage (Nham). Four treatments were prepared: a control without inoculation, a commercial probiotic culture (Starter V3), Lactobacillus plantarum TISTR 543, and Lactococcus lactis subsp. lactis TISTR 1520, with all cultures immobilized on cooked germinated rough rice. Physicochemical properties, including pH, total acidity, and water activity, as well as color, texture, GABA content, and microbial quality, were evaluated during fermentation at 30 °C and during storage at 4 °C. The results showed that probiotic starter cultures significantly enhanced GABA production, with Starter V3 yielding the highest levels, followed by L. plantarum TISTR 543. Probiotic inoculation accelerated acidification, resulting in consistently lower pH values and higher total acidity compared to the control, indicating more efficient fermentation. Lactic acid bacteria counts increased markedly during fermentation and remained high during storage, with L. plantarum TISTR 543 showing the strongest survival. Textural properties, particularly hardness and gumminess, improved in probiotic-supplemented samples, reflecting enhanced gel formation associated with protein and acid interactions. Although color attributes differed among treatments on the first day of fermentation, these differences diminished thereafter. All treatments met microbiological safety standards throughout fermentation and storage, with no detection of Staphylococcus aureus, Salmonella spp., or Escherichia coli. Overall, the findings confirm that probiotic starter cultures, especially Starter V3 and L. plantarum TISTR 543, can improve fermentation performance, enhance GABA accumulation, and support the development of safe, high-quality Nham with functional health-related potential.
References
Andrews, W. (1992). Manual of food quality control. 4. Rev. 1. Microbiological analysis. Food and Drug Administration. FAO Food and Nutrition paper, 14(4 Revis 1), 1-338.
Andrews, W. H., Jacobson, A., & Hammack, T. (2014). Salmonella (Ch. 5). In Bacteriological Analytical Manual (BAM) (8th ed.). Food and Drug Administration. Gaithersburg: AOAC International.
AOAC. (2000). Official Methods of Analysis of the AOAC. (17th ed.). Gaithersburg, MD, USA: The Association of Official Analytical Chemists International.
Bourne, M. C. (1978). Texture profile analysis. Food Technology, 32, 62-66.
Chen, C., & Chen, F. (2009). Study on the conditions to brew rice vinegar with high content of γ-amino butyric acid by response surface methodology. Food and Bioproducts Processing, 87(4), 334-340. https://doi.org/10.1016/j.fbp.2009.03.003
Chokesajjawatee, N., Pornaem, S., Zo, Y. G., Kamdee, S., Luxananil, P., Wanasen, S., & Valyasevi, R. (2009). Incidence of Staphylococcus aureus and associated risk factors in Nham, a Thai fermented pork product. Food Microbiology, 26(5), 547-551. https://doi.org/10.1016/j.fm.2009.02.009
Cui, Y., Miao, K., Niyaphorn, S., & Qu, X. (2020). Production of gamma-aminobutyric acid from lactic acid bacteria: A systematic review. International Journal of Molecular Sciences, 21(3), 995. https://doi.org/10.3390/ijms21030995
Dhakal, R., Bajpai, V. K., Baek, K. H., & Kang, S. C. (2012). Production of GABA (γ–aminobutyric acid) by microorganisms: A review. Brazilian Journal of Microbiology, 43(4), 1230-1241. https://doi.org/10.1590/S1517-83822012000400001
Gu, Y., Qiao, R., Jin, B., He, Y., & Tian, J. (2023). Effect of Limosilactobacillus fermentum 332 on physicochemical characteristics, volatile flavor components, and Quorum sensing in fermented sausage. Scientific Reports, 13(1), 3942. https://doi.org/10.1038/s41598-023-31161-2
Hwang, J., Kim, Y., Seo, Y., Sung, M., Oh, J., & Yoon, Y. (2023). Effect of starter cultures on quality of fermented sausages. Food Science of Animal Resources, 43(1), 1-9. https://doi.org/10.5851/kosfa.2022.e75
Jabeen, R., Jan, N., Naseer, B., Sarangi, P. K., Sridhar, K., Dikkala, P. K., Bhaswant, M., Hussain, S. Z., & Inbaraj, B. S. (2024). Development of germinated-brown-rice-based novel functional beverage enriched with γ-aminobutyric acid: nutritional and bio-functional characterization. Foods, 13(8), 1282. https://doi.org/10.3390/foods13081282
Jetwanna, P., Pilasombut, K., Limsupavanich, R., Sawatvivat, A., & Settakul, J. (2011). A study on quality and microbiology of beef Nham using Lactococcus lactis subsp. lactis P2 and Sb2 as a starter culture for fermentation. King Mongkut's Agricultural Journal, 29(3), 46-54.
Komatsuzaki, N., Tsukahara, K., Toyoshima, H., Suzuki, T., Shimizu, N., & Kimura, T. (2007). Effect of soaking and gaseous treatment on GABA content in germinated brown rice. Journal of Food Engineering, 78(2), 556-560. https://doi.org/10.1016/j.jfoodeng.2005.10.036
Krongkeha, W. (2022). Isolation and identification of GABA-producing lactic acid bacteria from fermented foods. RMUTSB Academic Journal. 10(1), 66-77.
Laranjo, M., Potes, M. E., & Elias, M. (2019). Role of starter cultures on the safety of fermented meat products. Frontiers in Microbiology, 10, 853. https://doi.org/10.3389/fmicb.2019.00853
Leroy, F., & De Vuyst, L. (2004). Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends in Food Science & Technology, 15(2), 67-78. https://doi.org/10.1016/j.tifs.2003.09.004
Li, X., Wang, H., Mehmood, W., Qian, S., Sun, Z., Zhang, C., & Blecker, C. (2018). Effect of storage temperatures on the moisture migration and microstructure of beef. Journal of Food Quality, 2018(1), 3873179. https://doi.org/10.1155/2018/3873179
Liu, Y., Gao, S., Cui, Y., Wang, L., Duan, J., Yang, X., Liu, X., Zhang, S., Sun, B., Yu, H., & Gao, X. (2024). Characteristics of lactic acid bacteria as potential probiotic starters and their effects on the quality of fermented sausages. Foods, 13(2), 198. https://doi.org/10.3390/foods13020198
Neffe-Skocińska, K., Okoń, A., Kołożyn‐Krajewska, D., & Dolatowski, Z. (2017). Amino acid profile and sensory characteristics of dry fermented pork loins produced with a mixture of probiotic starter cultures. Journal of the Science of Food and Agriculture, 97(9), 2953-2960. https://doi.org/10.1002/jsfa.8133
Meybodi, N., & Mortazavian, A. (2017). Probiotic supplements and food products: a comparative approach. Biochemistry& Pharmacology, 6(2), 2167-0501. https://doi.org/10.4172/2167-0501.1000227
Pakwan, C., Chitov, T., Chantawannakul, P., Manasam, M., Bovonsombut, S., & Disayathanoowat, T. (2020). Bacterial compositions of indigenous Lanna (Northern Thai) fermented foods and their potential functional properties. PLoS One, 15(11), e0242560. https://doi.org/10.1371/journal.pone.0242560
Pannerchelvan, S., Jawlan, L. L. L., Wasoh, H., Mohamed, M. S., Wong, F. W. F., Sobri, M. Z. M., Mohamad, R., & Halim, M. (2025). Enhancing cell viability and GABA production in fermented milk using fruit juice-coated alginate microencapsulated Lactiplantibacillus plantarum B7 during storage. International Microbiology, 28(7) 1857-1874. https://doi.org/10.1007/s10123-025-00662-7
Panya, A., Riebroy, S., Assavanig, A., Benjakul, S., & Visessanguan, W. (2002). Pale color formation in Nham, a Thai fermented pork sausage during fermentation. In Proceedings of the 4th Agro-Industry Conference. Bangkok, Thailand.
Phuapaiboon, P., Leenanon, B., & Levin, R. E. (2013). Effect of Lactococcus lactis immobilized within pineapple and yam bean segments, and jerusalem artichoke powder on its viability and quality of yogurt. Food and Bioprocess Technology, 6, 2751-2762. https://doi.org/10.1007/s11947-012-0940-4
Ratanaburee, A., Kantachote, D., Charernjiratrakul, W., & Sukhoom, A. (2013). Enhancement of γ-aminobutyric acid (GABA) in Nham (Thai fermented pork sausage) using starter cultures of Lactobacillus namurensis NH2 and Pediococcus pentosaceus
HN8. International Journal of Food Microbiology, 167(2), 170-176. https://doi.org/10.1016/j.ijfoodmicro.2013.09.014
Redruello, B., Saidi, Y., Sampedro, L., Ladero, V., Del Rio, B., & Alvarez, M. A. (2021). GABA-producing Lactococcus lactis strains isolated from camel’s milk as starters for the production of GABA-enriched cheese. Foods, 10(3), 633. https://doi.org/10.3390/ foods10030633
Santiyanont, P., Chantarasakha, K., Tepkasikul, P., Srimarut, Y., Mhuantong, W., Tangphatsornruang, S., Zo, Y. G., & Chokesajjawatee, N. (2019). Dynamics of biogenic amines and bacterial communities in a Thai fermented pork product Nham. Food Research International, 119, 110-118. https://doi.org/10.1016/j.foodres.2019.01.060
Singkhum, U., & Kangkun, W. (2021). Effects of natural colorants on physicochemical properties and sensory acceptance of Nham. International Journal of Agricultural Technology, 17(6), 2333-2350.
Sionek, B., Szydłowska, A., Trząskowska, M., & Kołożyn-Krajewska, D. (2024). The impact of physicochemical conditions on lactic acid bacteria survival in food products. Fermentation, 10(6), 298.
Suksai, U., & Chowwanapoonpohn, H. (2021). Factors associated with bacterial contamination in Nham and Pla-Som products. Thai Journal of Pharmacy Practice, 13(1), 251-264.
Swetwiwathana, A., & Visessanguan, W. (2015). Potential of bacteriocin-producing lactic acid bacteria for safety improvements of traditional Thai fermented meat and human health. Meat Science, 109, 101-105. https://doi.org/10.1016/j.meatsci.2015.05.030
Tangwatcharin, P., Nithisantawakhup, J., & Sorapukdee, S. (2020). Microbiological and physicochemical qualities of Moo Som (Traditional Thai fermented meat) inoculated with lactic acid bacteria starter. Walailak Journal of Science and Technology (WJST), 17(8), 788-800. https://doi.org/10.48048/wjst.2020.5222
TISI. (2004). Naem (Fermented ground pork), TIS 1219-2547. Bangkok, Thailand: Thailand Industrial Standard Institute.
Van Ba, H., Seo, H. W., Seong, P. N., Kang, S. M., Kim, Y. S., Cho, S. H., Park, B. Y., Ham, J. S., & Kim, J. H. (2018). Lactobacillus plantarum (KACC 92189) as a potential probiotic starter culture for quality improvement of fermented sausages. Korean Journal for Food Science of Animal Resources, 38(1), 189. https://doi.org/10.5851/kosfa.2018.38.1.189
Visessanguan, W., Benjakul, S., Panya, A., Kittikun, C., & Assavanig, A. (2005). Influence of minced pork and rind ratios on physico-chemical and sensory quality of Nham–a Thai fermented pork sausage. Meat Science, 69(2), 355-362. https://doi.org/10.1016/j.meatsci.2004.08.006
Visessanguan, W., Benjakul, S., Riebroy, S., & Thepkasikul, P. (2004). Changes in composition and functional properties of proteins and their contributions to Nham characteristics. Meat Science, 66(3), 579-588. https://doi.org/10.1016/S0309-1740(03)00172-4
Visessanguan, W., Benjakul, S., Smitinont, T., Kittikun, C., Thepkasikul, P., & Panya, A. (2006). Changes in microbiological, biochemical and physico-chemical properties of Nham inoculated with different inoculum levels of Lactobacillus curvatus. LWT-Food Science and Technology, 39(7), 814-826. https://doi.org/10.1016/j.lwt.2005.05.006
Woo, I. K., Hyun, J. H., Jang, H. J., Lee, N. K., & Paik, H. D. (2024). Probiotic Pediococcus acidilactici strains exert anti-inflammatory effects by regulating intracellular signaling pathways in LPS-induced RAW 264.7 cells. Probiotics and Antimicrobial Proteins, 1-12. https://doi.org/10.1007/s12602-024-10263-x
Woraprayote, W., Malila, Y., Sorapukdee, S., Swetwiwathana, A., Benjakul, S., & Visessanguan, W. (2016). Bacteriocins from lactic acid bacteria and their applications in meat and meat products. Meat Science, 120, 118-132. https://doi.org/10.1016/j.meatsci.2016.04.004
Xiudong, X., Ying, W., Xiaoli, L., Ying, L., & Jianzhong, Z. (2016). Soymilk residue (okara) as a natural immobilization carrier for Lactobacillus plantarum cells enhances soymilk fermentation, glucosidic isoflavone bioconversion, and cell survival under simulated gastric and intestinal conditions. PeerJ, 4, e2701. https://doi.org/10.7717/peerj.2701
Zhang, Y., Zhao, W., Lu, H., Yan, C., Yan, Q., Yao, X., Wang, W., Kang, D., & Liu, Y. (2023). Characteristics of microbial community in Linyi fermented pork sausage and their correlation with quality: Effects of the single Lactobacillus starter. LWT, 185, 115169. https://doi.org/10.1016/j.lwt.2023.115169
Zheng, Y., Zhao, G., Zhao, S., Li, X., Cui, W., Xu, L., Zhu C., & Tong, L. (2025). Don’t judge a sausage by its cover: Effects of inoculating three indigenous lactic acid bacteria on quality, moisture distribution, and protein structure in fermentation. Fermentation, 11(3), 134. https://doi.org/10.3390/fermentation11030134
Copyright (c) 2026 The Author
This work is licensed under a Creative Commons Attribution 4.0 International License.