The quality of robusta coffee (Coffea canephora) with the addition of Leuconostoc mesenteroides during wet fermentation
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Abstract. Rosaliana F, Swasti YR, Purwijantiningsih E. 2025. The quality of Robusta coffee (Coffea canephora) with the addition of Leuconostoc mesenteroides during wet fermentation. Biodiversitas 26: 3334-3342. Coffee consumption is inseparable from the health benefits of antioxidants in coffee beans. However, coffee contains caffeine, which affects the nervous system, kidneys, and blood vessels. Caffeine content is usually reduced using organic solvents and physical treatment. However, caffeine reduction using microorganisms, such as Lactic Acid Bacteria (LAB), is underexplored. This research aimed to improve the quality of robusta coffee by increasing its antioxidant activity and reducing caffeine content by adding Leuconostoc mesenteroides during wet fermentation. L. mesenteroides is a proteolytic bacterium that can degrade caffeine in coffee beans and enhance antioxidant activity. Robusta coffee has a higher caffeine content than Arabica. Therefore, the caffeine content needs to be reduced. The caffeine content was analysed using the HPLC method, and antioxidant activity was conducted using the DPPH method. The experimental design in this study was a completely randomized factorial design with two factors: fermentation time and the addition of L. mesenteroides. Each treatment has 3 replications. The data were analyzed using Analysis of Variance (ANOVA). The results show that the average caffeine content of espresso robusta green coffee beans was 1.26-2.19 mg/mL, while that of roasted coffee beans was 2.05-3.04%. Adding L. mesenteroides reduces the caffeine content of coffee beans by 13.34%. Espresso with green coffee beans has the average antioxidant activity ranging from 17.57±0.08%-23.32±0.08%, and espresso with roasted coffee beans ranges from 80.97±0.21%-84.87±0.08%; L. mesenteroides increases the antioxidant activity by 1.06%. In addition, the average moisture content of robusta green coffee beans fermented at 6, 12, and 18 hours with natural microbes and the addition of L. mesenteroides reached a moisture content of 9.82±0.10%-10.83±0.32%, while the roasted coffee beans ranged from 3.33±0.23%-3.80±0.16%. The average protein content of green coffee beans ranged from 10.51±0.76% to 13.28±0.67%, and roasted coffee beans from 11.38±0.88% to 12.55±0.50%. The total phenolic compound of green coffee beans ranged from 18.14±0.31 to 31.32±0.14 mg GAE/g, while the roasted beans reached 85.37±0.30 to 123.87±1.01 mg GAE/g. Adding L. mesenteroides reduces pH of green and roasted coffee beans. Testing other LABs on coffee beans can further explore these potential health benefits.
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References
Aguilar-Zarate P, Cruz-Hernandez MA, Montañez JC, Belmares-Cerda RE, Aguilar CN. 2014. Enhancement of tannase production by Lactobacillus plantarum CIR1: Validation in gas-lift bioreactor. Bioprocess Biosyst Eng 37 (11): 2305-2316. DOI: 10.1007/s00449-014-1208-3.
Alkaltham MS, Özcan MM, Uslu N, Salamatullah AM, Hayat K. 2020. Effect of microwave and oven roasting methods on total phenol, antioxidant activity, phenolic compounds, and fatty acid compositions of coffee beans. J Food Process Preserv 44 (11): e14874. DOI: 10.1111/jfpp.14874.
Choi S, Jung S, Ko KS. 2018. Effects of coffee extracts with different roasting degrees on antioxidant and anti-inflammatory systems in mice. Nutrients 10 (3): 363. DOI: 10.3390/nu10030363.
De Bruyn F, Zhang SJ, Pothakos V, Torres J, Lambot C, Moroni AV, Callanan M, Sybesma W, Weckx S, De Vuyst L. 2016. Exploring the impacts of postharvest processing on the microbiota and metabolite profiles during green bean production. Appl Environ Microbiol 83 (1): e02398-16. DOI: 10.1128/aem.02398-16.
de Jesus Cassimiro DM, Batista NN, Fonseca HC, Naves JAO, Coelho JM, Bernardes PC, Dias DR, Schwan RF. 2023. Wet fermentation of Coffea canephora by lactic acid bacteria and yeasts using the Self-Induced Anaerobic Fermentation (SIAF) method enhances the coffee quality. Food Microbiol 110: 104161. DOI: 10.1016/j.fm.2022.104161.
Diviš P, Po?ízka J, K?íkala J. 2019. The effect of coffee beans roasting on its chemical composition. Potravinarstvo Slovak J Food Sci 13 (1): 344-350. DOI: 10.5219/1062.
Elhalis H, Cox J, Frank D, Zhao J. 2020. The crucial role of yeasts in the wet fermentation of coffee beans and quality. Intl J Food Microbiol 333: 108796. DOI: 10.1016/j.ijfoodmicro.2020.108796.
Fessard A, Kapoor A, Patche J, Assemat S, Hoarau M, Bourdon E, Bahorun T, Remize F. 2017. Lactic fermentation as an efficient tool to enhance the antioxidant activity of tropical fruit juice and teas. Microorganism 5 (2): 23. DOI: 10.3390/microorganisms5020023.
Galanakis CM. 2017. Handbook of Coffee Processing By-Products. Elsevier, London.
George D, Mallery P. 2018. IBM SPSS Statistics 25 Step by Step: A Simple Guide and Reference, 15th eds. Routledge, New York. DOI: 10.4324/9781351033909.
Gokulakrishnan S, Chandraraj K, Gummadi SN. 2005. Microbial and enzymatic method for the removal of caffeine. Enzyme Microb Technol 37 (2): 225-232. DOI: 10.1016/j.enzmictec.2005.03.004.
Hanif M, Heru, Utami H. 2019. Variation of time and mass of spent coffee grounds on leaching oil from instant coffee residue. Inovasi Pembangunan: Jurnal Kelitbangan 7 (1): 49-61. DOI: 10.35450/jip.v7i1.119. [Indonesian]
Hasanah M, Maharani B, Munarsih E. 2017. Antioxidant power of robusta coffee leaf extract and fraction (Coffea robusta) against DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent. Indones J Pharm Sci Technol 4 (2): 42-49. DOI: 10.15416/ijpst.v4i2.10456. [Indonesian]
Higdon JV, Frei B. 2006. Coffee and health: A review of recent human research. Crit Rev Food Sci Nutr 46 (2): 101-123. DOI: 10.1080/10408390500400009.
Ibrahim I, Ayariga JA, Xu J, Boakai RK, Ajayi OS, Owusu-Kwarteng J. 2023. A comparative study of skimmed milk and cassava flour on the viability of freeze-dried lactic acid bacteria as starter cultures for yogurt fermentation. Foods 12: 1207. DOI: 10.3390/foods12061207.
Koduru L, Kim Y, Bang J, Lakshmanan M, Han NS, Lee D-Y. 2018. Genome-scale modeling and transcriptome analysis of Leuconostoc mesenteroides unravel the redox governed metabolic states in obligate heterofermentative lactic acid bacteria. Sci Rep 7 (1): 15721. DOI: 10.1038/s41598-017-16026-9.
Kostinek M, Specht I, Edward VA, Pinto C, Egounlety M, Sossa C, Mbugua S, Dortu C, Thonart P, Taljaard L, Mengu M, Franz CMAP, Holzapfel WH. 2007. Characterization and biochemical properties of predominant lactic acid bacteria from fermenting cassava for selection as starter cultures. Intl J Food Microbiol 114 (3): 342-351. DOI: 10.1016/j.ijfoodmicro.2006.09.029.
Kwak HS, Jeong Y, Kim M. 2018. Effect of yeast fermentation of green coffee beans on antioxidant activity and consumer acceptability. J Food Qual 2018 (1): 5967130. DOI: 10.1155/2018/5967130.
Latimer GW. 2023. Official Methods of Analysis of AOAC International. Oxford University Press, Oxford. DOI: 10.1093/9780197610145.001.0001.
Li X, Han Y, Wu X, Li L, Zhang R. 2023. Effects of inoculation with a binary mixture of Lactobacillus plantarum and Leuconostoc citreum on cell wall components of Chinese Dongbei suancai. Food Res Intl 173: 113458. DOI: 10.1016/j.foodres.2023.113458.
Liang N, Kitts DD. 2016. Role of chlorogenic acids in controlling oxidative and inflammatory stress conditions. Nutrients 8: 16. DOI: 10.3390/nu8010016.
Liao C, Tang X, Li M, Lu G, Huang X, Li L, Zhang M, Xie Y, Chen C, Li P. 2022. Effects of lactic acid bacteria, yeast, and their mixture on the chemical composition, fermentation quality, and bacterial community of cellulase-treated Pennisetum sinese silage. Front Microbiol 13: 1047072. DOI: 10.3389/fmicb.2022.1047072.
Liu Q, Zong C, Dong Z, Wu J, Zhu J, Li J, Zhang J, Shao T. 2020. Effects of cellulolytic lactic acid bacteria on the lignocellulose degradation, sugar profile and lactic acid fermentation of high-moisture alfalfa ensiled in low temperature seasons. Cellulose 27: 7955-7965. DOI: 10.1007/s10570-020-03350-z.
Liu SQ. 2016. Lactic acid bacteria: Leuconostoc spp. In: Reference Module in Food Science. National University of Singapore, Singapore.
Martins P, Jesus J, Santos S, Raposo LR, Roma-Rodrigues C, Baptista PV, Fernandes AR. 2015. Heterocyclic anticancer compounds: Recent advances and the paradigm shift towards the use of nanomedicine's toolbox. Molecules 20: 16852-16891. DOI: 10.3390/molecules200916852.
Mestanza M, Mori-Culqui PL, Chavez SG. 2023. Changes of polyphenols and antioxidants of arabica coffee varieties during roasting. Front Nutr 10: 1078701. DOI: 10.3389/fnut.2023.1078701.
Morales?Ramos V, Escamilla?Prado E, Ruiz?Carbajal RA, Pérez?Sato JA, Velázquez?Morales JA, Servín?Juárez R. 2020. On the soil?bean?cup relationships in Coffea arabica L. J Sci Food Agric 100 (15): 5434-5441. DOI: 10.1002/jsfa.10594.
Mousavi A, Pourakbar L, Moghaddam SS. 2022. Effects of malic acid and EDTA on oxidative stress and antioxidant enzymes of okra (Abelmoschus esculentus L.) exposed to cadmium stress. Ecotoxicol Environ Saf 248: 114320. DOI: 10.1016/j.ecoenv.2022.114320.
Naegele M. 2016. Determination of Caffeine in Coffee Products According to DIN 20481. Aglient Technologies Inc., Germany.
Nishiyama T, Hashiguchi Y, Sakata T, Sakaguchi T. 2003. Antioxidant activity of the fused heterocyclic compound 1,2,3,4-tetrahydroquinolines and related compounds-effect of ortho-substituents. Polym Degrad Stab 79 (2): 225-230. DOI: 10.1016/S0141-3910(02)00285-9.
Oestreich-Janzen S. 2010. Chemistry of coffee. In: Mander L (eds). Comprehensive Natural Products II Chemistry and Biology. Elsevier Ltd., Oxford.
Oliveira DM, Mota TR, Oliva B, Segato F, Marchiosi R, Ferrarese-Filho O, Faulds CB, Dos Santos WD. 2019. Feruloyl esterases: Biocatalysts to overcome biomass recalcitrance and for the production of bioactive compounds. Bioresour Technol 278: 408-423. DOI: 10.1016/j.biortech.2019.01.064.
Oliveira LS, Franca AS, Mendonça JCF, Barros-Júnior MC. 2006. Proximate composition and fatty acids profile of green and roasted defective coffee beans. LWT - Food Sci Technol 39 (3): 235-239. DOI: 10.1016/j.lwt.2005.01.011.
Park S-W, Park B-K, Ju Y-W, Benashvili M, Moon C-R, Lee S, Lee J, Son Y-J. 2025. Inoculation of starter cultures in dry processing enhanced the contents of bioactive compounds and sensory characteristics of Arabica coffee (Coffea arabica L.). Food Chem 475: 143226. DOI: 10.1016/j.foodchem.2025.143226.
Preedy VR. 2015. Coffee in Health and Disease Prevention. Elsevier Applied Science Publishers, London.
Purwoko T, Suranto, Setyaningsih R, Marliyana SD. 2022. Chlorogenic acid and caffeine content of fermented robusta bean. Biodiversitas 23 (2): 902-906. DOI: 10.13057/biodiv/d230231.
Purwoko T, Suranto, Setyaningsih R, Marliyana SD. 2023. Caffeine degradation by food microorganisms. Biodiversitas 24 (6): 3495-3502. DOI: 10.13057/biodiv/d240647.
Rao W, Fang Z, Chen Z, Wu J, Fang X. 2023. Antibacterial mechanism of metabolites of Leuconostoc mesenteroides against Serratia liquefaciens. LWT 187: 115335. DOI: 10.1016/j.lwt.2023.115335.
Reichembach LH, de Oliveira Petkowicz CL. 2020. Extraction and characterization of a pectin from coffee (Coffea arabica L.) pulp with gelling properties. Carbohydr Polym 245: 116473. DOI: 10.1016/j.carbpol.2020.116473.
Saanu AB. 2017. Pectinolytic activity of mutagenic strain of Leuconostoc Mesenteroides isolated from orange and banana fruit waste. J Appl Microbiol Biochem 2 (1): 1-6. DOI: 10.21767/2576-1412.100007.
Sari PDI, Swasti YR, Purwijantiningsih E. 2025. Antioxidant activity and quality of frozen yoghurt with Moringa oleifera Lamk. leaves extract. AgriTECH 45 (1): 31-40. DOI: 10.22146/agritech.83392.
Shen X, Wang Q, Wang H, Fang G, Li Y, Zhang J, Liu K. 2025. Microbial characteristics and functions in coffee fermentation: A review. Fermentation 11 (1): 5. DOI: 10.3390/fermentation11010005.
Sidoryk K, Jaromin A, Filipczak N, Cmoch P, Cybulski M. 2018. Synthesis and antioxidant activity of caffeic acid derivatives. Molecules 23 (9): 2199. DOI: 10.3390/molecules23092199.
Su H, Guo Y, Cheng H, Hu S, Zhang P, Yang Z. 2024. Probiotic and fermentation properties of Leuconostoc mesenteroides strain I1/53 from sugarcane juice by a multi-omics approach. LWT-Food Sci Technol 211: 116897. DOI: 10.1016/j.lwt.2024.116897.
Swasti YR, Leong LP, Purwijantiningsih E, Pranata FS. 2024. The effects of Lactobacillus plantarum addition to robusta coffee (Coffea canephora) during wet fermentation. Biodiversitas 25 (9): 3132-3140. DOI: 10.13057/biodiv/d250935.
Szlapinski SK, Charrette A, Guthrie N, Hilmas CJ. 2023. Paraxanthine safety and comparison to caffeine. Front Toxicol 5: 1117729. DOI: 10.3389/ftox 2023.1117729.
Tai E-S, Hsieh P-C, Sheu S-C. 2014. Effect of polygalacturonase and feruloyl esterase from Aspergillus tubingensis on demucilage and quality of coffee beans. Process Biochem 49 (8): 1274-1280. DOI: 10.1016/j.procbio.2014.05.001.
Tarigan IL, Ramadhan I, Pebridianto A, Latief M, Sutrisno S, Yusnaidar Y. 2023. Effect of fermentation on the quality of liberica coffee (Coffea Liberica) using Lactobacillus Plantarum and Leuconostoc Mesenteroides bacteria: Sensory analysis, volatile compound, antioxidant, and proximate analysis. SSRN. DOI: 10.2139/ssrn.4644595.
Utama CS, Zuprizal, Hanim C, Wihandoyo. 2018. Isolation and identification of lactic acid bacteria cellulolytic originated from fermented cabbage juice. Jurnal Aplikasi Teknologi Pangan 7 (1): 1-6. DOI: 10.17728/jatp.2155. [Indonesian]
Verster JC, Koenig J. 2018. Caffeine intake and its sources: A review of national representative studies. Crit Rev Food Sci Nutr 58 (8): 1250-1259. DOI: 10.1080/10408398.2016.1247252.
Wei F, Furihata K, Koda M, Hu F, Kato R, Miyakawa T, Tanokura M. 2012. 13C NMR-based metabolomics for the classification of green coffee beans according to variety and origin. J Agric Food Chem 60 (40): 10118-10125. DOI: 10.1021/jf3033057.
Wibowo KF, Swasti YR, Pranata FS. 2023. Low fat and high protein meat analogue of cowpea (Vigna unguiculata) with stabilizer cocoa pod husk extract (Theobroma cacao L.). Food Res 7 (5): 1-11. DOI: 10.26656/fr.2017.7(5).789.
Yang F, Chen C, Ni D, Yang Y, Tian J, Li Y, Chen S, Ye X, Wang L. 2023. Effects of fermentation on bioactivity and the composition of polyphenols contained in polyphenol-rich foods: A review. Foods 12 (17): 3315. DOI: 10.3390/foods12173315.
Zhang H, Huang-Fu H-P, Wang X, Zhao S-S, Liu Y, Lv H, Qin G-Y, Tan Z. 2021. Antibacterial activity of lactic acid producing Leuconostoc mesenteroides qz1178 against pathogenic Gallibacterium anatis. Front Vet Sci 8: 630294. DOI: 10.3389/fvets.2021.630294.
Zhang SJ, De Bruyn F, Pothakos V, Contreras GF, Cai Z, Moccand C, Weckx S, De Vuyst L. 2019. Influence of various processing parameters on the microbial community dynamics, metabolomic profiles, and cup quality during wet coffee processing. Front Microbiol 10: 2621. DOI: 10.3389/fmicb.2019.02621.