Production of dry culture from luwak-isolated Lacticaseibacillus paracasei using soy-based flour: its viability and activity

Authors

DOI:

https://doi.org/10.25186/.v21i.2458

Abstract

Luwak coffee is a unique coffee produced by the civet digestive system. Its rarity is a key characteristic, but this can be overcome by producing artificial luwak coffee using a dried culture of an isolate of Lacticaseibacillus paracasei, whose physiological properties have not been studied. This study aims to assess the physiology of the isolate and to characterize the luwak coffee-dry culture (LCDC), which various amounts of soy-based flour, including soybean, tempeh, and soybean sprouts, as a Nitrogen source. The stages include a study of the physiology of the isolate, and the addition of soy-based flour to the starter growth medium. The findings indicate that isolate cells are bacillus-shaped, gram-positive, facultative anaerobic, and homo-fermentative. They can grow at temperatures of 15 °C, 37 °C, and 45 °C, as well as at salt concentrations of 4 % and 6.5 %. LCDC supplemented with 10 g of tempeh flour showed promising results, with the highest viability (9.45 log10 CFU/g) and fermentation power, and a water content of less than 8%. As for fermentation capacity, the glucose fermentation capacity is 109.17 mg glucose (hg)-1 of LCDC, the total titratable acid is 5.11 mg LA (hg)-1, and the pH value is 4.30. Robusta luwak coffee, a specialty coffee, is produced by fermenting LCDC for 18 hours, achieving the highest total score of 81.38. The flavor profile is described as chocolate-like, with a hint of brown sugar, a spicy aftertaste, a sweet aftertaste, a hint of rubber, chocolate fruit, and a hint of medicine.

Key words: Specialty coffee; flavor profile; luwak coffee-dry culture; nitrogen sources; fermentation capacity.

References

Adadi, P. et al. (2024). Advancements in civet coffee production and analytical techniques: From aroma profiling to market dynamics and ethical considerations. Trends in Food Science & Technology, 154:104772.

Alvina, A. et al. (2004). Traditional tempe making process. Halal Food Scientific Journal, 1(1):9-12.

Association of Official Analytical Chemists - AOAC. (2005). Official methods of analysis. association of official analytical chemists. (18th Edition), Washington: Benjamin Franklin Station.

Arfianty, B. N. et al. (2017). Dynamics of bacterial population and total acid in bekasam fermentation of patin fish (Pangasius hypopthalmus). Scientific Journal of Experimental Biology and Biodiversity, 4(2):43-49.

Aryal, S. (2022). Lactobacilli MRS Broth- Composition, Principle, Preparation, Results, Uses, Microbe Notes. Available in: <https://microbenotes.com/lactobacilli-mrs-broth/>.

Astawan, M. et al. (2016). Physicochemical characteristics of sprouted soybean tempeh flour. Jurnal Gizi dan Pangan, 11(1):35-42.

Azizah, N. et al. (2019). Effect of the concentration of lactic acid bacteria Lactobacillus plantarum and Lactobacillus casei on the microbiological and chemical quality of probiotic mayonnaise. Journal of Animal Science, 18(2):79-85.

Badan Standar Nasional Indonesia - BSNI. (2009). Maximum Limits of Microbial Contamination in Food. 7388:2009. Available in: <https://kupdf.net/download/sni-7388-2009_5c83567ae2b6f5ea44dbed4e_pdf>.

Bengoa, A. A. et al. (2019). Physicochemical, immunomodulatory and safety aspects of milks fermented with Lactobacillus paracasei isolated from kefir. Food Research International, 123:48-55.

Bengoa, A. A. et al. Health-promoting properties of Lacticaseibacillus paracasei: A focus on kefir isolates and exopolysaccharide-producing strains. Foods, 10(10):2239.

Budiharjo, R. et al. (2017). Effect of NaCl concentration on specific extracellular protease activity and growth of halophilic bacteria isolate from Madura salt pond bittern. Journal of Chemical Science and Applications, 20(3):142-145.

Burgé, G. et al. (2015). Relationships between the use of Embden Meyerhof pathway (EMP) or Phosphoketolase pathway (PKP) and lactate production capabilities of diverse Lactobacillus reuteri strains. Journal of Microbiology, 53:702-710.

Charalampopoulos, D. et al. (2002). Growth studies of potentially probiotic lactic acid bacteria in cereal-based substrates. Journal of Applied Microbiology, 92(5):851-859.

Cao, X. et al. (2023). Effects of postharvest processing on aroma formation in roasted coffee–a review. International Journal of Food Science and Technology, 58(3):1007-1027.

Colautti, A. F. et al. (2024). Investigating safety and technological traits of a leading probiotic species: Lacticaseibacillus paracasei. Nutrients, 16:2212.

Dishan, A., & Gönülalan, Z. (2024). Lacticaseibacillus paracasei AD22 stress response in brined white cheese matrix: In vitro probiotic profiles and molecular characterization, Probiotics and Antimicrobial Proteins, 17:1725-1738.

de Melo Pereira, G. V. et al. (2020). Lactic acid bacteria: What coffee industry should know?. Current Opinion in Food Science, 31:1-8.

Ediviani, W. et al. (2014). Effectiveness of powder and solid ragi (starter) from Dago (Bandung, West Java, Indonesia) traditional market on production of bioethanol from rice straw. Makara Journal of Science, 18(1):19-25.

Elhalis, H. et al. (2020). Ecological diversity, evolution and metabolism of microbial communities in the wet fermentation of Australian coffee beans. International Journal of Food Microbiology, 321:108544.

Fadilah, U. et al. (2018). Study of the effect of initial ph of media and fermentation duration on the ethanol production process from jackjack seed flour hydrolysate using Saccharomycess Cerevisiae. Jurnal Rekayasa Dan Manajemen Agroindustri, 6(2):92-102.

Fauzi, M. et al. (2017). Chemical characteristics of artificial luwak robusta coffee fermented by luwak culture and α-amylase. Jurnal Penelitian Pascapanen Pertanian, 14(3):144-153.

Fauzi, M. et al. (2023). Identification of lactic acid bacteria isolated from developed dried coffee starter culture used as a fermentation agent to produce Robusta civet coffee. Biodiversitas, 24(7):3715-3722.

Finanda, A. et al. (2021). Isolation and characterization of a genus of lactic acid bacteria from fermented flesh of kepok bananas (Musa paradisiaca L.). Protobiont Journal, 10(2):37-41.

Febrianto, N. A., & Zhu, F. (2023). Coffee bean processing: Emerging methods and their effects on chemical, biological and sensory properties. Food Chemistry, 412:135489.

Fitriyah, D. et al. (2020). Analysis of the nutritional content of rice from several transgenic rice lines Pac Nagdong/Ir36. Artery: Journal of Health Sciences, 1(2):154-160.

Hartoyo, A., & Sunandar, F. H. (2006). Utilization of composite flour of white sweet potatoes (Ipomea batatas L.), Germinated soybean (Glycine max Merr), and germinated mung bean (Virginia radiate L) as wheat flour partial substituent of alternative food, high protein energy biiscuits). Jurnal Teknologi dan Indusri Pangan, 17(1):50-57.

Hill, D. et al. (2018). The Lactobacillus casei Group: History and health related applications. Frontiers in Microbiology, 9:2107.

Ifmalinda, I. et al. (2019). Chemical characteristics comparison of palm civet coffee (Kopi Luwak) and arabica coffee beans. Journal of Applied Agricultural Science and Technology, 3(2):280-288.

Kesuma, I. P. E. A. W. et al. (2021). Isolation and screening of cellulolytic bacteria in civet feces. Nandur, 1(3):139-147.

König, H., & Fröhlich, J. (2017). Lactic acid bacteria. In H. König., G. Unden., & J. Fröhlich, J. (eds). Biology of Microorganisms on Grapes, in Must and in Wine. Springer, Cham. (pp.3-41).

Kurnia, M. et al. (2020). Isolation and identification of lactic acid bacteria from traditional food of the rejang tribe in Bengkulu province: "Lemea." Allotropes, 4(1):25-32.

Lamoureux, A. (2022). Kopi luwak, the expensive delicacy known as ‘cat poop coffee’. Ed. John Kuroski. Available in: <https://allthatsinteresting.com/kopi-luwak-cat-poop-coffee>.

Lee, L. W. et al. (2015). Coffee fermentation and flavor-An intricate and delicate relationship. Food Chemistry, 185:182-191.

Lewis-Whelan, B. et al. (2023). Welfare and management of civets in civet coffee tourism plantations. Journal of Applied Animal Welfare Science, 27(3):561-574.

Lingle, T. R., & Menon, S. N. (2017). Chapter 8 - Cupping and grading-discovering character and quality. In B. Folmer. The craft and science of coffee. Academic Press, (pp.181-203).

Liu, L. et al. (2023). Transcriptomics of Lactobacillus paracasei: Metabolism patterns and cellular responses under high-density culture conditions. Frontiers in Bioengineering and Biotechnology, 11:1274020.

Maidannyk, V. A. et al. (2022). The effect of high protein powder structure on hydration, glass transition, water sorption, and thermomechanical properties. Foods, 11(3):292.

Mustakim., Yusmarini., & Herawati, N. (2016). The use of corn flour and tempeh flour in making crackers. Jom Faperta, 3(2):1-15.

Muzaifa, M. et al. (2016). Kopi Luwak: Produksi, kualitas, dan masalah. Syiah Kuala University Press.

Nurdini A. L. et al. (2015). Microbial growth dynamics during tempeh fermentation in two different home industries. International Food Research Journal, 22(4):1668-1674.

Ozogul, F. et al. (2022). Lactic acid bacteria: Lactobacillus acidophilus. In P. L. H. McSweeney., & J. P. McNamara. Encyclopedia of dairy sciences. Academic Press: Cambridge, MA, USA, (pp.187-197).

Patterson, E. et al. (2020). Lacticaseibacillus paracasei Lpc-37® improves psychological and physiological markers of stress and anxiety in healthy adults: A randomized, double-blind, placebo-controlled and parallel clinical trial (the Sisu study). Neurobiology of Stress, 13:100277.

Pinillos-Miñano, R. M. et al. (2022). Isolation of lactic acid bacteria from the feces of ring-tailed coati (Nasua nasua), Biochemical and fermentative aspects related to coffee ermentation. Applied Biochemistry and Microbiology, 58:S102S112.

Prastujati, A. U. et al. (2018). Effect of starter concentration on alcohol content, ph, and total titratable acid (TAT) of kefir whey. Journal of Applied Animal Science, 1(2):63-69.

Putrama, W. I. G. P. et al. (2022). Characteristics of lactic acid bacteria in civet feces in several areas of the island of Bali. Medika Udayana, 11(7):69-73..

Rahmawati, L. et al. (2020). Inovasi pembuatan cookies kaya gizi dengan proporsi tepung bekatuldan tepung kedelai. Jurnal Agrotek Ummat, 7(1):30-36.

Rohman, A. R. et al. (2019). Effect of fermentation time on acidity, total lactic acid bacteria, total yeast and hedonic quality green coconut (cocos nucifera) water kefir. Jurnal Teknologi Pangan, 3(1):127-133.

Safrida, Y. D. (2021). Uji total plate count (TPC) bakteri pada minuman teh poci homemade di gampong batoh banda aceh. Jurnal Serambi Engineering, 6(2):1790-1796.

Specialty coffee association - SCA. (2022). Coffee sensory and cupping Handbook. Specialty coffee association. Available in: <https://sca.coffee/store/p/coffee-sensory-cupping-handbook>.

Sholihah, R. et al. (2017). Formulation of snakehead fish meal (channa striata), soybean sprout flour (Glycine max merr) and corn sprout flour (Zea mays) for instant cereal for malnourished toddlers. Indonesian Journal of Health Information, 3(2):132-144.

Silva, L. C. et al. (2024). Enhancing sensory quality of coffee: The impact of fermentation techniques on Coffea arabica cv. Catiguá MG2. Foods, 13(5):653.

Sionek, B. et al. (2024). The impact of physicochemical conditions on lactic acid bacteria survival in food products. Fermentation, 10(6):298.

Subagio, A. et al. (2004). Characterization of myofibrillar protein from goldband goat fish (Upeneus moluccensis) and Bigeye Scad Fish (Selar crumenophthalmus). Jurnal Teknologi dan Industri Pangan, 15(1):70-78.

Sudarmadji. S. et al. (2010). Food and agricultural analysis. (2nd ed). Liberty, Yogyakarta. 125p.

Taufik, Y. et al. (2017). Effect of concentration of fruit pulp and soybean flour (Glycine max) on the fit bar characteristics of black mulberry (Morus nigra L.). Pasundan Food Technology Journal, 5(1):10-17.

Torres-Miranda, A. et al. (2022). Characterization and identification of probiotic features in Lacticaseibacillus paracasei using a comparative genomic analysis approach. Probiotics and Antimicrobial Proteins, 14:1211-1224.

Wang, Y. et al. (2022). Metabolism characteristics of lactic acid bacteria and the expanding applications in food industry. Frontiers in Bioengineering and Biotechnology, 12(9):612285.

Wikandari, R. et al. (2021). Correlations between the chemical, microbiological characteristics and sensory profile of fungal fermented food. Fermentation, 7(4):261.

Wu, Y. et al. (2025). Analysis of flavor quality differences between LZ coffee and coffee based on untargeted metabolomics. Food Bioscience, 68:106514.

Yulia, R. et al. (2016). Analisis kadar kafein kopi luwak dengan variasi jenis kopi, spesies luwak dan cara pengolahan dengan metoda TLC scanner. Sain Farm Klinik, 2(2):171-175.

Downloads

Published

2026-06-12

How to Cite

FAUZI, Mukhammad; PRIHANDINI, Winda Almira; JAYUS; AFRILIANA, Asmak; HARSONO, Soni Sisbudi; BAKRI, Abi; QUINTAO, Gregorio. Production of dry culture from luwak-isolated Lacticaseibacillus paracasei using soy-based flour: its viability and activity. Coffee Science - ISSN 1984-3909, [S. l.], v. 21, p. e212458, 2026. DOI: 10.25186/.v21i.2458. Disponível em: https://coffeescience2.ufla.br/index.php/Coffeescience/article/view/2458. Acesso em: 20 oct. 2026.