Enhanced Cellulose Production in Kombucha SCOBY Through Microbial and Genetic Optimization

Authors

  • Hussaini Adib Haslan Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
  • Murni Halim Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
  • Fadzlie Wong Faizal Wong Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
  • Zulfazli M. Sobri Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
  • Nor'Aini Abdul Rahman Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia
  • Mohd Sabri Pak-Dek Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
  • Yanty Noorzianna Abdul Manaf Halal Research Group, Faculty of Food Science and Nutrition, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia.
  • Helmi Wasoh Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

DOI:

https://doi.org/10.54987/jemat.v13i1.1110

Keywords:

Cellulose, Kombucha, Biosynthesis, Fermentation, Acetic Acid Bacteria

Abstract

The symbiotic culture of bacteria and yeast (SCOBY) represents a dynamic microbial consortium that plays a fundamental role in kombucha fermentation. This complex system consists of acetic acid bacteria (AAB), lactic acid bacteria (LAB), and various yeast species whose synergistic interactions generate bioactive compounds including organic acids, polyphenols, and bacterial cellulose (BC). Within the SCOBY consortium, Komagataeibacter and Gluconobacter spp. (AAB) catalyze the oxidative conversion of ethanol to acetic acid, generating an acidic microenvironment that both inhibits competing microorganisms and promotes bacterial cellulose biosynthesis. LAB, including Lactobacillus and Pediococcus, enhance fermentation stability, probiotic potential, and biofilm structure through exopolysaccharide production and bacteriocin secretion. Yeasts like Saccharomyces cerevisiae and Zygosaccharomyces bailii metabolize sugars into ethanol and CO₂, supporting AAB activity and contributing to flavor complexity. Recent advances in biosynthesis research have identified over 200 microbial species in SCOBY, with high-throughput sequencing revealing key metabolic pathways. Genetic optimization of BC production involves the bcsABCD operon, which regulates cellulose synthase activity, with CRISPR and metabolic engineering enhancing yield and crystallinity (84-89%). Engineered strains of Komagataeibacter xylinus demonstrate improved BC properties, including nanofibrillar density (2-4 nm) and water retention (>99%). However, SCOBY’s industrial application faces challenges, including batch variability, environmental sensitivity, and inconsistent microbial profiles, necessitating precision fermentation with defined consortia for standardized production. Future research should focus on robust clinical validation of health claims and scalable bioprocessing techniques to harness SCOBY’s full potential in food, biotechnology, and biomedical applications.

References

Adnan A, Sudin ASNZ. The state of play: symbiotic culture of bacteria and yeasts (SCOBY) in textile industry. J Teknologi (Sci Eng). 2024;86(3):175-86.

Harrison K, Curtin C. Microbial composition of SCOBY starter cultures used by commercial kombucha brewers in North America. Microorganisms. 2021;9(5):1060.

Liao T, Li XR, Fan L, Zhang B, Zheng WM, Hua JJ, et al. Nature of back slopping kombucha fermentation process: insights from the microbial succession, metabolites composition changes and their correlations. Front Microbiol. 2024;15.

Laavanya D, Shirkole S, Balasubramanian P. Current challenges, applications, and future perspectives of SCOBY cellulose of kombucha fermentation. J Clean Prod. 2021;295:126454.

Su J, Tan Q, Tang Q, Tong Z, Yang M. Research progress on alternative kombucha substrate transformation and the resulting active components. Front Microbiol. 2023;14(14):1254014.

Molina-Ramírez C, Rojas O, Gómez C, Zuluaga R, Castro C, Osorio M, et al. Effect of different carbon sources on bacterial nanocellulose production and structure using the low pH resistant strain Komagataeibacter medellinensis. Materials (Basel). 2017;10(6):639.

Feng X, Ge Z, Wang Y, Xia X, Zhao B, Dong M. Production and characterization of bacterial cellulose from kombucha-fermented soy whey. Food Prod Process Nutr. 2024;6:20.

Prajapati K, Prajapati J, Patel D, Patel R, Varshnei A, Saraf M, et al. Multidisciplinary advances in kombucha fermentation, health efficacy, and market evolution. Arch Microbiol. 2024;206(9).

May A, Narayanan S, Alcock J, Varsani A, Maley C, Aktipis A. Kombucha: a novel model system for cooperation and conflict in a complex multi-species microbial ecosystem. PeerJ. 2019;7:e7565.

Yuliana E, Permana AH. Isolation and characterization of microorganisms from SCOBY (Symbiotic Culture of Bacteria and Yeast) during kombucha fermentation. WARTA AKAB. 2022;46(2):18-22

Yang S, Bai M, Kwok LY, Zhong Z, Sun Z. The intricate symbiotic relationship between lactic acid bacterial starters in the milk fermentation ecosystem. Crit Rev Food Sci Nutr. 2023;65(4):728-45.

Biswas S, Biswas S, Sarkar U, Mitra A. Insights into the challenges and resolutions in the bacterial fermentation process. Amsterdam: Elsevier; 2024.

Rafay R, Allegue T, Fowler SJ, Rodríguez J. Exploring the limits of carbohydrate conversion and product formation in open mixed culture fermentation. J Environ Chem Eng. 2022;10(3):107513.

Healy LE, Zhu X, Kakagianni M, Poojary MM, Sullivan C, Tiwari U, et al. Fermentation of brown seaweeds Alaria esculenta and Saccharina latissima for new product development using Lactiplantbacillus plantarum, Saccharomyces cerevisiae and kombucha SCOBY. Algal Res. 2023;76:103322.

Kaashyap M, Mantri N, Cohen M. Microbial diversity and characteristics of kombucha as revealed by metagenomic and physicochemical analysis. Nutrients. 2021;13(12):4446.

Laureys D, Aerts M, Vandamme P, De Vuyst L. The buffer capacity and calcium concentration of water influence the microbial species diversity, grain growth, and metabolite production during water kefir fermentation. Front Microbiol. 2021;12:628599.

Chen C, Liu BY. Changes in major components of tea fungus metabolites during prolonged fermentation. J Appl Microbiol. 2000;89(5):834-9.

Girard M, Gullo M, Ren H. A systematic, complexity-reduction approach to dissect the microbial and functional diversity of kombucha. eLife. 2023;12:e76401.

Cruz MA, Flor-Unda O, Avila A, Garcia MD, Cerda-Mejía L. Advances in bacterial cellulose production: a scoping review. Coatings. 2024;14(11):1401.

Wang B, Naren N, Mutukumira AN, Rutherfurd-Markwick K, Zhang XX. Microbiological and physico-chemical characteristics of black tea kombucha fermented with a New Zealand starter culture. Foods. 2023;12(12):2314.

Dartora B, Sant’Anna V, Hickert LR, Fensterseifer M, Ayub MAZ, Flôres SH, et al. Factors influencing kombucha production: effects of tea composition, sugar, and SCOBY. Food Sci Technol. 2023.43, e8123,

Van TP, Pham GB, Quang HP, Do AD, Thi HTT, Phan QK, et al. Multi-strain probiotics enhance the bioactivity of cascara kombucha during microbial composition-controlled fermentation. Prev Nutr Food Sci. 2023;28(4):502-13.

Venegas C, Saona L, Urbina K, Quintrel P, Peña T, Mardones W, et al. Addition of Saccharomyces eubayanus to SCOBY fermentations modulates the chemical and volatile compound profiles in kombucha. Food Microbiol. 2023;116:104357.

Chong A, Chin N, Talib R, Basha R. Modelling pH dynamics, SCOBY biomass formation, and acetic acid production of kombucha fermentation using black, green, and oolong teas. Processes. 2024;12(7):1301.

Kim G, Baek K, Lee G, Lee J, Moon J, Seo S. Development of starter cultures for precision fermentation of kombucha with enriched gamma-aminobutyric acid (GABA) content. Fermentation. 2025;11(1):17.

Lin SP, Calvar IL, Catchmark JM, Liu JR, Demirci A, Cheng KC. Biosynthesis, production and applications of bacterial cellulose. Cellulose. 2013;20(5):2191-219.

Revin V, Liyaskina E, Nazarkina M, Bogatyreva A, Shchankin M. Cost-effective production of bacterial cellulose using acidic food industry by-products. Braz J Microbiol. 2018;49:151-9.

Costa AFS, Almeida FCG, Vinhas GM, Sarubbo LA. Production of bacterial cellulose by Gluconacetobacter hansenii using corn steep liquor as nutrient source. Front Microbiol. 2017;8:2027.

Li Z, Wang L, Hua J, Jia S, Zhang J, Liu H. Production of nano bacterial cellulose from wastewater of candied jujube-processing industry using Acetobacter xylinum. Carbohydr Polym. 2015;120:115-9.

Yao S, Hao L, Zhou R, Jin Y, Huang J, Wu C. Multispecies biofilms in fermentation: biofilm formation, microbial interactions, and communication. Compr Rev Food Sci Food Saf. 2022;21(4):3346-75.

Abol-Fotouh D, Hassan MA, Shokry H, Roig A, Azab MS, Kashyout AEHB. Bacterial nanocellulose from agro-industrial wastes: low-cost and enhanced production by Komagataeibacter saccharivorans MD1. Sci Rep. 2020;10(1):3491.

Gorgieva S, Trček J. Bacterial cellulose: production, modification and perspectives in biomedical applications. Nanomaterials (Basel). 2019;9(10):1352.

Jayalakshmi T, Gayathry G, Kumutha K, Sabarinathan KG, Amutha R, Veeramani P. Plausible avenues and applications of bioformulations from symbiotic culture of bacteria and yeast. J Pure Appl Microbiol. 2024;18(3):1489-501.

Neffe-Skocińska K, Sionek B, Ścibisz I, Kołożyn-Krajewska D. Acid contents and the effect of fermentation condition of kombucha tea beverages on physicochemical, microbiological and sensory properties. CyTA J Food. 2017;15(4):601-7.

Mangayil R, Rajala S, Pammo A, Sarlin E, Luo J, Santala V, et al. Engineering and characterization of bacterial nanocellulose films as low cost and flexible sensor material. ACS Appl Mater Interfaces. 2017;9(22):19048-56.

Gullo M, La China S, Falcone PM, Giudici P. Biotechnological production of cellulose by acetic acid bacteria: current state and perspectives. Appl Microbiol Biotechnol. 2018;102(16):6885-98.

Semjonovs P, Ruklisha M, Paegle L. Cellulose synthesis by Komagataeibacter rhaeticus strain P 1463 isolated from kombucha. Appl Microbiol Biotechnol. 2016;100(13):5565-76.

Içen H, Corbo MR, Sinigaglia M, Korkmaz BIO, Bevilacqua A. Microbiology and antimicrobial effects of kombucha, a short overview. Food Biosci. 2023;56:103270.

Wang S, Li S, Ji C, Lin X, Liang H, Qi L, et al. Effect of synthetic microbial community on nutraceutical and sensory qualities of kombucha. Int J Food Sci Technol. 2020;55(10):3327-33.

Fabricio MF, Vargas BK, Tischer B, Wagner R, Ribeiro SR, Cordeiro N, et al. Revamping kombucha production: achieving consistency and probiotic potential through a tailor-made microbial consortium. Int J Gastron Food Sci. 2023;34:100844.

Buldum G, Mantalaris A. Systematic understanding of recent developments in bacterial cellulose biosynthesis at genetic, bioprocess and product levels. Int J Mol Sci. 2022;22(13):7192.

Abidi W, Torres-Sánchez L, Siroy A, Krasteva PV. Weaving of bacterial cellulose by the Bcs secretion systems. FEMS Microbiol Rev. 2021;46(2):fuab051.

Serra DO, Hengge R. Cellulose in bacterial biofilms. Berlin: Springer; 2019.

Römling U, Galperin MY. Bacterial cellulose biosynthesis: diversity of operons, subunits, products, and functions. Trends Microbiol. 2015;23(9):545-57.

Amr A, Ibrahim H. Bacterial cellulose: biosynthesis and applications. London: IntechOpen; 2023.

Avcioglu NH. Bacterial cellulose: recent progress in production and industrial application. World J Microbiol Biotechnol. 2022;38(5).

Krystynowicz A, Czaja W, Wiktorowska-Jezierska A, Gonçalves-Miśkiewicz M, Turkiewicz M, Bielecki S. Factors affecting the yield and properties of bacterial cellulose. J Ind Microbiol Biotechnol. 2002;29(4):189-95.

El-Gendi H, Taha HT, Ray JB, Saleh AK. Recent advances in bacterial cellulose: a low-cost effective polymer. Cellulose. 2022;29(5):7495-7533.

Kumar R, Singh OV, Singh S. Bioconversion of lignocellulosic biomass: biochemical and molecular perspectives. J Ind Microbiol Biotechnol. 2008;35(5):377-91.

Ryngajłło M, Jędrzejczak-Krzepkowska M, Kubiak K, Ludwicka K, Bielecki S. Towards control of cellulose biosynthesis by Komagataeibacter using systems-level and strain engineering strategies: current progress and perspectives. Appl Microbiol Biotechnol. 2020;104(15):6565-85.

Downloads

Published

31.07.2025

Issue

Section

Articles

How to Cite

Enhanced Cellulose Production in Kombucha SCOBY Through Microbial and Genetic Optimization. (2025). Journal of Environmental Microbiology and Toxicology, 13(1), 39-46. https://doi.org/10.54987/jemat.v13i1.1110