Previous Article in Journal
Biosynthesis of the Siderophore Desferrioxamine E in Rouxiella badensis SER3 and Its Antagonistic Activity Against Fusarium brachygibbosum
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Applied Microbiology
Volume 5
Issue 3
10.3390/applmicrobiol5030092
applmicrobiol-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Review

Bacterial Cellulose Production in Co-Culture Systems: Opportunities, Challenges, and Future Directions

by
Dheanda Absharina
1,*,
Filemon Jalu Nusantara Putra
2,
Chiaki Ogino
2,
Sándor Kocsubé
1,
Csilla Veres
1 and
Csaba Vágvölgyi
1,*
1
Department of Biotechnology and Microbiology, Faculty of Science and Informatics, University of Szeged, Közép Fasor, H-6726 Szeged, Hungary
2
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-Ku, Kobe 657-8501, Japan
*
Authors to whom correspondence should be addressed.
Appl. Microbiol. 2025, 5(3), 92; https://doi.org/10.3390/applmicrobiol5030092
Submission received: 15 July 2025 / Revised: 9 August 2025 / Accepted: 12 August 2025 / Published: 26 August 2025
Browse Figures
Versions Notes

Abstract

Bacterial cellulose (BC), a nanostructured biopolymer produced by Komagateibacter spp., exhibits remarkable mechanical strength, purity, and biocompatibility, making it highly attractive for applications in biomedicine, food, and sustainable materials. Despite its potential, monoculture fermentation suffers from low yield and limited scalability. This review highlights the innovative application of co-culture fermentations as a novel strategy, where Komagataeibacter is paired with complementary microorganisms such as yeasts, lactic acid bacteria, and photosynthetic microbes. This approach has emerged as a promising solution to overcome the limitations of monoculture by enhancing BC productivity, tailoring material properties, and improving sustainability. We explore the synergistic interactions within co-cultures, including metabolic cross-feeding and in situ polymer integration, while also addressing critical challenges such as microbial stability and operational complexity. Unlike previous reviews focused primarily on BC biosynthesis, applications, or genetic engineering, this article emphasizes co-culture fermentation with Komagataeibacter as a novel and underexplored strategy to improve the yield, functionality, and scalability of BC production.

1. Introduction

Bacterial cellulose (BC) is a highly pure extracellular polysaccharide predominantly synthesized by species of the genus Komagataeibacter (formerly classified as Acetobacter or Gluconacetobacter) [1,2]. Although chemically identical to plant-derived cellulose, composed of linear β-1,4-linked D-glucopyranose units, BC exhibits superior physicochemical and structural characteristics due to its microbial origin and distinct biosynthetic pathway [1,2,3]. The key distinction lies in its exceptional purity; unlike plant cellulose, which is embedded within a lignocellulosic matrix containing lignin, hemicellulose, and pectin, BC is secreted in an essentially pure form, typically exceeding 99% cellulose content [3,4]. Structurally, BC features an ultrafine three-dimensional nanofiber network with fiber diameters ranging from 20 to 100 nm—approximately 100 times thinner than plant-derived microfibrils—resulting in a very high surface-area-to-volume ratio [2,3,4]. This nanostructure contributes to its high crystallinity (70–80%), high degree of polymerization (DP ~2000–6000), and outstanding mechanical properties, including its excellent tensile strength and Young’s modulus [3,4]. BC hydrogels can absorb up to 100 times their dry weight in water, a property dramatically enhanced through composite formation. For instance, BC/poly-AEM composites exhibit swelling capacities up to ~6200% by preventing collapse of the BC structure during drying, while BC/acrylic hydrogel composites reach 4000–6000% swelling and, in vivo, significantly promote burn healing via enhanced epithelialization and fibroblast proliferation [3,5,6,7]. Furthermore, BC is inherently biocompatible, non-toxic, and biodegradable—qualities essential for biomedical applications such as wound dressings, drug delivery systems, and tissue engineering scaffolds [5,6,7,8,9,10,11,12]. The cellulose synthesis process, in which fibers form tightly packed ribbons under standard conditions and become more loosely organized with additive supplementation, is illustrated in Figure 1.
BC biosynthesis in Komagataeibacter xylinus, a model high-yield strain, is mediated by the cellulose synthase complex encoded by the bcs operon. This complex catalyzes glucose polymerization into glucan chains that self-assemble into nanofibrils [6,7,10,11]. While prior reviews have focused on BC’s structural and physicochemical features [3,4,9], biomedical applications [5,6,7,8,11,12], or advances in nanocellulose and cellulose biosynthesis at the genetic level [9,10,13], few have addressed co-culture fermentation strategies involving Komagataeibacter. This review fills that gap by synthesizing emerging evidence on co-culture systems, particularly those pairing Komagataeibacter with yeasts, lactic acid bacteria, or photosynthetic organisms, and highlights how such systems offer a promising, underexplored approach for scalable and sustainable BC production.

2. The Co-Culture Paradigm: A Strategy for Enhanced Yield and Functionalization of BC

2.1. Overcoming BC Monoculture Limitations Through Co-Culture Strategies

Conventional monoculture-based BC production, though foundational, faces persistent challenges related to scalability, productivity, and functional customization. The most common method, static cultivation, yields high-quality BC pellicles at the air–liquid interface but is hindered by slow kinetics, poor oxygen diffusion, and labor-intensive operations, thus limiting industrial viability [2,8,9,10]. Agitated or submerged bioreactor systems improve oxygen and nutrient transfer and offer potential for scale-up [8,14]. However, these systems may cause shear-induced defects, lower crystallinity, and increased formation of non-cellulose-producing (Cel−) mutants, ultimately reducing BC quality and yield [2,4,15].
Meanwhile, co-culture fermentation leverages synergistic interactions between Komagataeibacter and partner microbes to improve yield, expand substrate range, and functionalize BC [16,17,18,19,20]. Mechanisms include metabolic cross-feeding, pH regulation, and dynamic interspecies interactions [16,19,20]. For example, ethanol produced by yeasts can serve as an additional carbon source for Komagataeibacter, while organic acids from lactic acid bacteria (LAB) stimulate BC biosynthesis [20]. Such systems also enhance process robustness and adaptability under industrial conditions.
Nonetheless, uniform pellicle thickness remains a challenge especially in static systems due to oxygen and nutrient gradients that result in heterogeneous structures [14,18,21]. Applications such as biomedical scaffolds or textile membranes require mechanically robust, uniform BC layers [22]. Additionally, co-cultures introduce complexity: differing oxygen requirements and nutrient competition can destabilize microbial balance and hinder BC formation [23]. However, they also enable division of labor and facilitate in situ incorporation of microbial exopolymers—such as pullulan or hyaluronic acid (HA)—into the BC matrix, yielding composites with enhanced functionality and mechanical properties [24,25,26,27,28].

2.2. Bioreactor Configurations for Co-Culture-Based BC Fermentation

Recent innovations in bioreactor design have expanded the applicability of co-culture strategies by addressing oxygenation, shear stress, and microbial spatial organization. The following bioreactor types have been employed in co-culture-based BC production:
  • Static Fermentation
Reported that 181 studies employed a static system for monoculture fermentation [21], whereas its application in co-culture settings has been far less common. Typically, shallow trays where Komagataeibacter interacts with yeasts or LAB at the air–liquid interface. Metabolite exchange enhances pellicle formation and crystallinity, though productivity remains low due to surface area limitations and slow kinetics [10,15,29].
2.
Agitation Fermentation
Reported in 33 studies [21], this approach uses shaking flasks or stirred-tank reactors to enhance mixing, oxygenation, and substrate dispersion. Shear forces may impair BC structure, favoring amorphous forms. Strategies like intermittent agitation or optimized stirring speeds aim to reduce such effects [10,30,31].
3.
Airlift Bioreactor
Gas-lift systems provide high oxygen transfer in low-shear environments—suitable for aerobic co-cultures. Wu and Li (2015) achieved high-yield BC production using a modified airlift design [32]. However, BC pellet formation and excessive sparging may disrupt synthesis [10,29,32].
4.
Rotating Biological Contactor (RBC)
RBC systems utilize disks partially submerged in a culture medium, allowing microorganisms to cyclically access air and nutrients, which supports aerobic and facultative anaerobic co-cultures such as Komagataeibacter with yeasts or fungi [10,29,33]. These systems offer high oxygen transfer, low shear stress, and continuous biofilm growth. A novel rotating disk bioreactor (RDBR) introduced by Pilafidis et al. (2025) featured perforated horizontal disks, achieving superior oxygenation and nutrient diffusion. The system reported 15.2 g/L BC yield and 2.17 g/L/day productivity with improved fibril architecture [34].
5.
Dual-Vessel Co-Culture Reactors
These systems spatially separate the aerobic (e.g., Komagataeibacter) and anaerobic/microaerophilic (e.g., LAB or yeast) components into two connected vessels. Nutrient and metabolite exchange is facilitated through a closed-loop circulation system. While highly customizable and effective in maintaining microbial balance, dual-vessel systems introduce higher capital costs and engineering complexity. They are particularly suited for bioprocesses requiring strict metabolic separation [10,29,35].
6.
Mesh Dispenser Vessel (MDV) Bioreactor
The MDV integrates static pellicle growth with intermittent nutrient feeding. A mesh scaffold suspends the growing BC at the air–liquid interface, allowing thick, vertically expanding pellicles without submersion. Loh et al. (2025) reported pellicle thicknesses exceeding 80 mm, with a 3.4-fold increase in glucose conversion and improved water-use efficiency [2,36]. This reactor is well-suited for co-culture systems requiring sustained aeration and minimal disturbance, though it demands precise control of feeding regimens and harvesting techniques.
Figure 2 represents the different bioreactor types which have been employed in co-culture-based BC production, followed by a brief overview of each bioreactor type.

3. Advancements in BC Production: From Natural Consortia to Co-Culture Fermentation Systems

The conceptual foundation for co-culture strategies in BC production originates from traditional fermented foods, which rely on complex, naturally occurring symbiotic communities of bacteria and yeasts, commonly referred as SCOBY (symbiotic culture of bacteria and yeast), to form cellulosic pellicles. Well-known examples include kombucha (fermented tea), in which yeasts such as Saccharomyces and Zygosaccharomyces hydrolyze sucrose into glucose and fructose, producing ethanol as a byproduct [16,24]. Acetic acid bacteria (Komagataeibacter spp.) subsequently utilize both sugars and ethanol to synthesize BC and organic acids [16,18,37,38]. Similarly, in nata de coco (fermented coconut water), Komagataeibacter nataicola coexists with lactic acid bacteria (Lactobacillus spp.), leading to improved BC yields [18,24].
Early research predominantly focused on isolating Komagataeibacter monocultures to optimize BC biosynthesis [38,39]. However, limitations soon emerged, including inefficient substrate utilization, byproduct inhibition (e.g., gluconic acid accumulation), and restricted yields [28,38]. In recent years, BC production has shifted toward engineered co-culture systems designed to deliberately optimize microbial interactions. A landmark study by Seto et al. [17] demonstrated that co-culturing Gluconacetobacter xylinus with Lactobacillus mali increased BC yield threefold compared to monoculture. Their work provided one of the first clear validations that a well-designed co-culture can achieve higher performance than a single strain. This transition from relying on undefined natural consortia to utilizing rationally constructed co-cultures marks a significant step forward in BC biotechnology. It enables precise control over microbial interactions, allowing researchers to enhance yields, customize material properties, and expand substrate versatility. The progress of co-culture BC fermentation studies is summarized in Table 1.

3.1. Key Advances in Co-Culture BC Production by Komagataeibacter

Numerous studies have demonstrated that pairing Komagataeibacter with compatible microbes can substantially enhance BC yield and tailor its physicochemical properties. In metabolic cross-feeding systems, one partner microbe produces metabolites that directly stimulate BC biosynthesis. For example, co-culturing K. xylinus with the ethanol-producing yeast Kluyveromyces marxianus on brewing yeast waste resulted in a twofold yield increase compared to monoculture [24,35]. In another dual-bacterial system, Bacillus cereus metabolized corn stover hydrolysate and secreted acetoin and 2,3-butanediol, both of which stimulated K. xylinus to produce more cellulose than in monoculture [17].
Notably, co-culture advantages extend beyond BC yield improvements. They also facilitate the use of low-cost feedstocks (e.g., agricultural residues, brewery waste) by combining Komagataeibacter with microbes capable of degrading complex nutrients [44]. Beyond microbial pairing, strain selection within the genus Komagataeibacter itself significantly influences co-culture outcomes. While K. xylinus is the most commonly engineered strain, less explored species such as K. rhaeticus, K. oboediens, and K. medellinensis may possess adaptive traits, such as acid stress tolerance or oxygen efficiency, that are particularly advantageous under co-culture conditions [14,20]. Incorporating these strains could unlock novel efficiencies in co-culture BC production.
Besides co-culture strategies, in situ BC functionalization has been explored by pairing Komagataeibacter with exopolysaccharide-producing microbes. For example, co-culturing K. hansenii with Aureobasidium pullulans produces BC–pullulan nanocomposites with ~22% higher yield and improved tensile strength and Young’s modulus [45]. Similarly, engineered Lactococcus lactis secreting hyaluronic acid (HA) can be paired with G. hansenii [46,47], producing BC/HA composites with greater fiber width, higher water retention, and improved elasticity, properties valuable for wound dressings [6,7,12,46]. Such in situ integration of functional polymers reduces the need for purified additives, and post-processing fermentation promotes homogeneous incorporation of functional polymers into the BC structure, as demonstrated in L. lactis–HA and A. pullulans–pullulan co-culture systems, reducing production costs and simplifying workflows [45,46,47]. A comprehensive comparison of co-culture systems is presented in Table 2.

3.2. Key Challenge of Co-Culture Fermentation-Based BC Production

Despite the promise of co-culture systems, several unresolved issues limit their industrial application. These include long-term stability in continuous fermentation, the controllability of population dynamics, unintended consequences for BC quality, and the genetic instability of Komagataeibacter strains [10,14,21,51,52]. Furthermore, the broader scientific debate emphasizes skepticism about the feasibility of co-cultures in industrial contexts. Despite their synergistic potential, co-culture systems often experience population imbalance due to nutrient competition, pH sensitivity, and differences in growth rates [53,54,55]. This challenge becomes particularly acute in continuous or fed-batch processes at industrial scale, where fluctuating conditions can destabilize mutualistic interactions [54,55]. In addition, co-culture optimization requires careful tuning of inoculum ratios, medium composition, and inoculation timing [4,8,10,15,21]. Nutrient-enriched media often required for partner microbes may also increase the contamination risk, leading to yield variability and altered BC physicochemical properties [53,56]. Addressing these challenges will require system designs, advanced real-time monitoring, and deeper mechanistic understanding of interspecies interactions [57,58]. Table 3 summarizes the key challenges, representative studies, and potential mitigation strategies.
Meanwhile, Figure 3 presents representative studies along with potential mitigation strategies for these challenges and opportunities.

4. Industrial Co-Culture Fermentation: Lessons and Comparison with BC System

In industrial biotechnology, co-culture systems are extensively employed in processes such as biohydrogen production, ethanol fermentation, and bioplastic synthesis to improve substrate utilization and product yields. For example, in biofuel production, facultative anaerobes (e.g., Klebsiella, Streptococcus) are co-cultured with obligate anaerobes (Clostridium spp.), enabling the degradation of complex substrates and the removal of inhibitory byproducts (e.g., H2, formate), thereby enhancing overall metabolic performance and process stability [30,62]. These consortia exemplify metabolic division of labor, where each organism specializes in a specific step of a pathway. A similar approach could be adapted for BC fermentation, for instance, by pairing cellulolytic Clostridium species with Komagataeibacter to valorize lignocellulosic waste streams into BC [63].
For instance, Li et al. (2023) reported that co-culturing K. xylinus with B. cereus on corn stover hydrolysate significantly increased BC yield. B. cereus fermented complex sugars into acetoin and 2,3-butanediol, which stimulated BC biosynthesis [17]. This illustrates how co-culture can broaden substrate scope (e.g., agro-industrial residues) and enhance productivity through metabolic complementarity.
Additional insights come from traditional fermented foods that rely on stable mixed cultures. In yogurt production, defined LAB co-cultures (e.g., Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) co-metabolize lactose and synergistically produce flavor compounds. Kefir represents a complex microbial ecosystem, comprising diverse LAB (e.g., L. kefiranofaciens, L. kefiri, Lactiplantibacillus plantarum) and yeasts (e.g., Saccharomyces cerevisiae, Kluyveromyces marxianus) embedded in kefiran-rich grains. These microbes cooperate metabolically by converting lactose and other sugars into organic acids, ethanol, and bioactive compounds, stabilizing the community and enhancing functionality [64,65].
In nata de coco, a traditional fermented coconut water product, BC is synthesized through synergistic interactions between K. nataicola and Lactobacillus spp. [18,43]. This bacterial–bacterial co-culture exemplifies metabolic complementarity, where lactic acid bacteria produce metabolites that modulate pH and stimulate cellulose biosynthesis [17,18,46,53,54,66]. A similar but distinct consortium appears in kombucha fermented on Sargassum fusiforme (brown algae), where Zygosaccharomyces dominates throughout, and the bacterial profile shifts from Acetobacter to Komagataeibacter over time [52]. This shift correlates with acid accumulation (final pH ~2.77), increased antioxidant activity, and volatile metabolite production (e.g., ethanol, acetone, ethyl acetate).
Kombucha, a fermented tea, demonstrates another example of well-orchestrated microbial synergy: yeasts such as Saccharomyces and Zygosaccharomyces hydrolyze sucrose into glucose and fructose and ferment them into ethanol and CO2, which are subsequently utilized by Komagataeibacter spp. to produce acetic acid and bacterial cellulose, forming the characteristic SCOBY biofilm [16,24,45,66]. High-throughput sequencing of commercial SCOBYs revealed Brettanomyces and Komagataeibacter as predominant members, with Zygosaccharomyces, Lachancea, and Starmerella functionally compensating when Brettanomyces is scarce [45]. These communities cluster into four reproducible “archetypes,” each representing a distinct yeast–bacteria balance that maintains cellulose production [51,66].

5. Techno-Economic Analysis of BC Production: Monoculture vs. Co-Culture

5.1. Cost Considerations in BC Production: Monoculture vs. Co-Culture

The economic feasibility of BC production remains a key barrier to industrial adoption, primarily due to the high cost of culture media and fermentation infrastructure. Zhong et al. (2020) identified two principal strategies to lower costs: enhancing BC yield and utilizing cost-effective nutrient sources, as media can constitute up to 30% of total production expenses [10,14]. Conventional monoculture systems using Komagataeibacter spp. in a Hestrin–Schramm (HS) medium yield high-purity BC but are hindered by low yields and costly inputs, particularly refined sugars, yeast extract, and peptones [8,10,14,60,61]. To overcome these limitations, considerable research has focused on replacing costly substrates with agro-industrial byproducts. Promising alternatives such as glycerol [21], corn steep liquor [67], citrus pulp water [68], molasses and cashew apple juice [69], vinegar residue [70], coconut water [61,71], and biodiesel-derived residues [31] have demonstrated cost reduction of up to 97% while promoting circular bioeconomy goals [31,71]. However, scalability remains limited by feedstock variability and seasonal availability.
Henry et al. (2024) reported that substituting an HS medium with solid-state fermentation (SSF)-treated rice bran (RB) and cereal dust reduced BC production costs by up to 90%, from AUD 17.08/g to AUD 2.51/g [72]. Recent findings further support the feasibility of low-cost substrates. Nóvak et al. (2024) reported that a black tea and sugar medium yielded the lowest cost (USD 0.10/L), compared to HS medium (USD 1.36/L), while enzymatically treated substrates reached up to USD 5.58/L [73]. These findings reinforce the viability of SSF-treated agro-waste as a scalable, economical solution for BC production.

5.2. Techno-Economic Analysis and Industrial Feasibility of Co-Culture-Based BC Production

Techno-economic analysis (TEA) and life cycle assessment (LCA) are essential tools for evaluating feasibility of scaling BC production. These models assess parameters including capital expenditure, operating costs, resource efficiency, and profitability, offering strategic insight into process optimization and cost reduction pathways [61,71]. Although current TEA studies remain limited particularly for co-culture systems, existing analyses reveal promising economic scenarios when alternative substrates, reactor innovations, and microbial consortia are integrated. A notable example is the kombucha-based BC production system, involving a symbiotic co-culture of Komagataeibacter and osmophilic yeasts. Behera et al. (2022) simulated a 60-ton/year production facility using SuperPro Designer®, estimating a total capital investment of USD 13.72 million and annual operating costs of USD 3.8 million [61]. A comprehensive techno-economic assessment estimated the production cost of dry bacterial cellulose at USD 63.8 per kilogram, with a capital recovery period of 4.2 years, a return on investment (ROI) of 23.64%, and an internal rate of return (IRR) of 16.48%. A SuperPro Designer® simulation by Dourado et al. (2016) modeled a large-scale facility producing 504 tons/year, showing that use of beet molasses instead of refined sugars reduced the production cost to USD 14.8/kg. This model estimated a capital investment of USD 13 million, with operating costs at USD 7.4 million/year and a projected net profit of USD 3.3 million/year [71]. These results emphasize the significant impact of media composition, particularly carbon and nitrogen sources, which may account for 30–40% of total production costs [71].
Notably, approximately 89% of the total operating costs were attributed to infrastructure and labor, highlighting the relatively minor economic contribution of raw materials such as a sweet tea medium [31]. These data highlight advancements in automated, modular bioreactor systems capable of continuous or semi-continuous operation. Automation not only reduces labor costs but also ensures product consistency. Real-time monitoring tools with dynamic feedback controls for pH, dissolved oxygen, and substrate concentration are essential to optimize fermentation conditions and scale-up potential [21,31,71,73].
El-Gendi et al. (2022) emphasized that effective bioprocess design integrating media optimization, advanced reactor engineering, and microbial performance enhancement is central to achieving economically and environmentally sustainable BC production [15]. Furthermore, Islam et al. (2017) underscored that bioreactor design directly influences BC yield and cost-effectiveness [31].
Market dynamics further influence TEA outcomes. High-purity BC can command prices up to USD 120/kg in biomedical markets, while bulk food-grade BC (e.g., nata de coco) is sold for USD 0.2–1.0/kg in Southeast Asia [61]. These low-cost fermentations often rely on open-vat methods with coconut water under non-sterile conditions. However, Sundaram et al. (2023) demonstrated that glycerol–sesame seed meal hydrolysate significantly increased economic productivity and cost efficiency compared to an HS medium (USD 6.98 vs. 4.34 per dollar of nutrient per day), highlighting the value of waste valorization in co-culture fermentation [29]. Key techno-economic indicators for alternative media cultivation are summarized in Table 4.

6. Omics Integration for Co-Culture Engineering

Omics technologies, including genomics, transcriptomics, proteomics, and metabolomics, are increasingly applied in co-culture systems to unravel microbial interactions and their impact on metabolism, phenotypes, and product quality.
Transcriptomic profiling facilitates omics integration in co-culture systems, allowing researchers to reveal dynamic gene expression changes in mixed cultures compared to monocultures. By analyzing global mRNA expression patterns, it becomes possible to determine which genes in Komagataeibacter are up- or downregulated in response to metabolites secreted by partner strains or environmental modifications. An omics-based study by Wang et al. (2018) identified distinct gene expression profiles in Komagataeibacter under different carbon sources and co-substrate conditions [74]. In a related study, Fei et al. (2023) found that adding exopolysaccharides from E. coli into Gluconacetobacter culture significantly improved the tensile strength of the cellulose pellicle [43]. Proteomic studies revealed upregulation of enzymes and stress-related pathways under ethanol-enriched co-culture, linking microbial interaction to enhanced BC synthesis [43,55]. The end-to-end workflow for sample preparation, MS acquisition, and multivariate analysis is summarized in Figure 4.
These findings underscore omics as essential tools for optimizing fermentation, improving material properties, and guiding the development of next-generation BC-based products. For instance, natural consortia such as kefir grains composed of LAB, acetic acid bacteria, and yeasts embedded in exopolysaccharide matrices efficiently coordinate metabolic pathways [64]. Comparative genomics by Ryngajłło et al. (2019) revealed that certain Komagataeibacter strains possess multiple bcs operons and adaptive stress genes, e.g., K. medellinensis carries four copies of the type II operon, offering genetic plasticity [75].
Synthetic systems mimic such consortia. Liu et al. (2021) engineered a co-culture of Synechococcus elongatus (photoautotroph) and E. coli (heterotroph), where S. elongatus fixed CO2 and light, while E. coli produced isoprene. The system extended fermentation from 100 to 400 h, and increased isoprene yield eightfold. Multi-omics revealed oxidative stress responses in E. coli, triggered by reactive oxygen species (ROS), which activated protective pathways and extended viability [76].
Co-cultures have also boosted industrial enzyme production. For example, co-culturing Bacillus amyloliquefaciens with UV/ARTP-mutated TGase-producing strains enhanced enzyme activity to 16.91 U/mL—a 22.71% increase—through improved protein secretion and stress adaptation. Genomic analysis identified 95 mutations affecting codon usage, while transcriptomics revealed 470 differentially expressed genes related to metabolism, sporulation, and enzyme activation [77].
Omics also uncover cryptic biosynthetic gene clusters (BGCs). Co-culturing Micromonospora with Rhodococcus activated the silent keyicin gene cluster via interspecies signaling, revealed through transcriptomic and proteomic analyses [78]. Metabolomics similarly exposes latent biosynthetic capacity: Eurotium amstelodami co-cultured with Bacillus spp. enhanced anti-S. aureus activity through altered metabolic pathways and novel secondary metabolites [79].
In continuous microbial cultures, metabolomics-guided co-culture strategies offer solutions to key bottlenecks like contamination, strain degeneration, and metabolic instability. Co-culturing photoautotrophs with heterotrophs can establish stable micro-ecological systems that support prolonged and efficient biosynthesis. For instance, Chlorella vulgaris and E. coli co-cultivation for isoprene production yielded a tenfold increase (0.6 g/L) and extended fermentation from 100 to 350 h. C. vulgaris supported E. coli growth and isoprene yield, while E. coli modulated algal physiology, forming a mutualistic relationship driven by oxidative stress. Transcriptomic data showed elevated expression of antioxidant (CysK, CysE, SerA, AhpC, AhpF) and repair system genes (YtfE, NfuA, YebG) in E. coli. Interestingly, C. vulgaris upregulated cysteine biosynthesis, suggesting cross-feeding to mitigate ROS [80]. Collectively, multi-omics strategies enable precise engineering of microbial consortia for consistent, high-yield BC production, unlocking their full industrial potential.

7. Conclusions and Future Perspectives

This review highlights the emerging potential of Komagataeibacter-based co-culture systems as a sustainable alternative to monoculture fermentation for BC production. Through engineered microbial interactions, co-cultures enhance BC yield, enable the utilization of diverse agro-industrial residues, and facilitate in situ functionalization of the cellulose matrix. Advances in bioreactor configurations, including dual-vessel systems and rotating disk bioreactors, have improved oxygenation and scalability, addressing critical limitations of static cultures. Multi-omics approaches have further elucidated the metabolic networks and regulatory pathways underpinning co-culture performance, providing a foundation for rational strain engineering and process optimization. Collectively, these developments position co-culture fermentation as a scalable and resource-efficient platform for BC biomanufacturing.
Future efforts should focus on the rational design of defined microbial consortia guided by systems biology and omics-integrated modeling to enhance metabolic complementarity and process stability. Underexplored Komagataeibacter species with stress-resilient phenotypes warrant further investigation for their compatibility in co-culture systems. From a techno-economic perspective, the adoption of modular, automated bioreactors and the integration of low-cost, regionally available substrates are essential to reduce production costs and improve economic viability. Moreover, adaptive laboratory evolution and synthetic regulatory circuits offer promising avenues to enhance co-culture robustness under industrial conditions. Establishing standardized cultivation protocols, quality control metrics, and regulatory frameworks will be critical to support the commercialization of co-culture-derived BC across biomedical, food, and sustainable material sectors.

Author Contributions

Conceptualization, D.A. and C.V. (Csaba Vágvölgyi); methodology, D.A., F.J.N.P. and C.V. (Csilla Veres); software, S.K.; validation, C.V. (Csilla Veres) and C.O.; formal analysis, D.A. and C.V. (Csaba Vágvölgyi); investigation, D.A. and C.V. (Csaba Vágvölgyi); resources, D.A. and C.V. (Csaba Vágvölgyi); data, S.K., F.J.N.P. and C.V. (Csilla Veres); writing—original draft preparation, D.A., F.J.N.P., C.O., S.K., C.V. (Csilla Veres) and C.V. (Csaba Vágvölgyi); writing—review and editing, D.A. and C.V. (Csaba Vágvölgyi); visualization, D.A. and S.K.; supervision, C.V. (Csaba Vágvölgyi); project administration, C.V. (Csilla Veres) and C.V. (Csaba Vágvölgyi); funding acquisition, D.A. and C.V. (Csaba Vágvölgyi). All authors have read and agreed to the published version of the manuscript.

Funding

The authors express their gratitude for the financial support provided by the Stipendium Hungaricum program (2022_479964; 2022–2027).

Acknowledgments

The authors declare that no individuals are acknowledged in this section; therefore, consent was not required.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Girard, V.-D.; Chaussé, J.; Vermette, P. Bacterial Cellulose: A Review of Production, Applications and Perspectives. J. Appl. Polym. Sci. 2024, 141, e55163. [Google Scholar] [CrossRef]
  2. Nguyen, V.T.; Flanagan, B.; Gidley, M.J.; Dykes, G.A. Characterization of Cellulose Production by a Gluconacetobacter xylinus Strain from Kombucha. Curr. Microbiol. 2008, 57, 449–453. [Google Scholar] [CrossRef]
  3. Sozcu, S.; Frajova, J.; Wiener, J.; Venkataraman, M.; Tomkova, B.; Militky, J. Synthesis of Acetobacter xylinum Bacterial Cellulose Aerogels and Their Effect on the Selected Properties. Gels 2025, 11, 272. [Google Scholar] [CrossRef] [PubMed]
  4. Jozala, A.F.; Pértile, R.A.N.; dos Santos, C.A.; Santos-Ebinuma, V.C.; Seckler, M.M.; Gama, F.M.; Pessoa, A., Jr. Bacterial Cellulose Production by Gluconacetobacter xylinus Employing Alternative Culture Media. Appl. Microbiol. Biotechnol. 2015, 99, 1181–1190. [Google Scholar] [CrossRef] [PubMed]
  5. Melro, L.; Alves, C.; Fernandes, M.; Rocha, S.; Mehravani, B.; Ribeiro, A.I.; Azevedo, S.; Cardoso, V.F.; Carvalho, Ó.; Dourado, N.; et al. Bacterial Nanocellulose as a Versatile Scaffold for Biomedical Applications: Synthesis, Functionalization, and Future Prospects. Appl. Mater. Today 2025, 46, 102858. [Google Scholar] [CrossRef]
  6. Mohamad, N.; Mohd Amin, M.C.; Pandey, M.; Ahmad, N.; Rajab, N.F. Bacterial Cellulose/Acrylic Acid Hydrogel Synthesized via Electron Beam Irradiation: Accelerated Burn Wound Healing in an Animal Model. Carbohydr. Polym. 2014, 114, 312–320. [Google Scholar] [CrossRef]
  7. Figueiredo, A.R.P.; Figueiredo, A.G.P.R.; Silva, N.H.C.S.; Barros-Timmons, A.; Almeida, A.; Silvestre, A.J.D.; Freire, C.S.R. Antimicrobial Bacterial Cellulose Nanocomposites Prepared by In Situ Polymerization of 2-Aminoethyl Methacrylate. Carbohydr. Polym. 2015, 123, 443–453. [Google Scholar] [CrossRef]
  8. Fernandes, I.A.A.; Pedro, A.C.; Ribeiro, V.R.; Bortolini, D.G.; Ozaki, M.S.C.; Maciel, G.M.; Haminiuk, C.W.I. Bacterial Cellulose: From Production Optimization to New Applications. Int. J. Biol. Macromol. 2020, 164, 2598–2611. [Google Scholar] [CrossRef]
  9. Singhania, R.R.; Patel, A.K.; Tsai, M.-L.; Chen, C.-W.; Dong, C.D. Genetic Modification for Enhancing Bacterial Cellulose Production and Its Applications. Bioengineered 2021, 12, 6793–6807. [Google Scholar] [CrossRef]
  10. Cruz, M.A.; Flor-Unda, O.; Avila, A.; Garcia, M.D.; Cerda-Mejía, L. Advances in Bacterial Cellulose Production: A Scoping Review. Coatings 2024, 14, 1401. [Google Scholar] [CrossRef]
  11. Öz, Y.E.; Bingül, N.D.; Morçimen, Z.G.; Şendemir, A.; Hameş, E.E. Fabrication of porous bone scaffolds using degradable and mouldable bacterial cellulose. Cellulose 2024, 31, 2921–2935. [Google Scholar] [CrossRef]
  12. Czaja, W.K.; Young, D.J.; Kawecki, M.; Brown, R.M., Jr. The future prospects of microbial cellulose in biomedical applications. Biomacromolecules 2007, 8, 1–12. [Google Scholar] [CrossRef]
  13. Hur, D.H.; Choi, W.S.; Kim, T.Y.; Lee, S.Y.; Park, J.H.; Jeong, K.J. Enhanced Production of Bacterial Cellulose in Komagataeibacter xylinus via Tuning of Biosynthesis Genes with Synthetic RBS. J. Microbiol. Biotechnol. 2020, 30, 1430–1435. [Google Scholar] [CrossRef]
  14. Zhong, C. Industrial-scale production and applications of bacterial cellulose. Front. Bioeng. Biotechnol. 2020, 8, 605374. [Google Scholar] [CrossRef]
  15. El-Gendi, H.; El-Sayed, E.S.R.; Hassan, M.L.; Hebeish, A. Recent advances in bacterial cellulose: A low-cost effective production media, optimization strategies and applications. Cellulose 2022, 29, 7495–7533. [Google Scholar] [CrossRef]
  16. Shi, S.; Wei, Y.; Lin, X.; Liang, H.; Zhang, S.; Chen, Y.; Dong, L.; Ji, C. Microbial metabolic transformation and antioxidant activity evaluation of polyphenols in Kombucha. Food Biosci. 2023, 51, 102287. [Google Scholar] [CrossRef]
  17. Li, W.; Huang, X.; Liu, H.; Lian, H.; Xu, B.; Zhang, W.; Sun, X.; Wang, W.; Jia, S.; Zhong, C. Improvement in Bacterial Cellulose Production by Co-Culturing Bacillus cereus and Komagataeibacter xylinus. Carbohydr. Polym. 2023, 313, 120892. [Google Scholar] [CrossRef]
  18. Jiang, H.; Song, Z.; Hao, Y.; Hu, X.; Lin, X.; Liu, S.; Li, C. Effect of co-culture of Komagataeibacter nataicola and selected Lactobacillus fermentum on the production and characterization of bacterial cellulose. LWT Food Sci. Technol. 2023, 173, 114224. [Google Scholar] [CrossRef]
  19. Lin, X.; Song, Z.; Jiang, H.; Hao, Y.; Hu, X.; Liu, S.; Li, C. Production of bacterial cellulose in the medium with yeasts pre-fermented coconut water or with addition of selected amino acids. Foods 2022, 11, 3627. [Google Scholar] [CrossRef]
  20. Seto, A.; Saito, Y.; Matsushige, M.; Kobayashi, H.; Sasaki, Y.; Tonouchi, N.; Tsuchida, T.; Yoshinaga, F.; Ueda, K.; Beppu, T. Effective cellulose production by a coculture of Gluconacetobacter xylinus and Lactobacillus mali. Appl. Microbiol. Biotechnol. 2006, 73, 915–921. [Google Scholar] [CrossRef]
  21. Quijano, L.; Rodrigues, R.; Fischer, D.; Tovar-Castro, J.D.; Payne, A.; Navone, L.; Hui, Y.; Yan, H.; Pinmanee, P.; Poon, E.; et al. Bacterial Cellulose Cookbook: A Systematic Review on Sustainable and Cost-Effective Substrates. J. Bioresour. Bioprod. 2024, 9, 379–409. [Google Scholar] [CrossRef]
  22. Fernandes, M.; Souto, A.P.; Dourado, F.; Gama, M. Application of bacterial cellulose in the textile and shoe industry: Development of biocomposites. Polysaccharides 2021, 2, 566–581. [Google Scholar] [CrossRef]
  23. Absharina, D.; Padri, M.; Veres, C.; Vágvölgyi, C. Bacterial cellulose: From biofabrication to applications in sustainable fashion and vegan leather. Fermentation 2025, 11, 23. [Google Scholar] [CrossRef]
  24. Paronyan, M.; Saghatelyan, L.; Avetisyan, S.; Koloyan, H.; Kinosyan, M.; Bagiyan, V.; Hovhannisyan, S.; Akopian, O.; Hovsepyan, A. Co-Cultivation of Komagataeibacter xylinus MS2530 with Various Yeast Strains: Production and Characterization of Bacterial Cellulose Films. Carbohydr. Polym. Technol. Appl. 2025, 11, 100840. [Google Scholar] [CrossRef]
  25. Peng, X.-Y.; Wu, J.-T.; Shao, C.-L.; Li, Z.-Y.; Chen, M.; Wang, C.-Y. Co-culture: Stimulate the metabolic potential and explore the molecular diversity of natural products from microorganisms. Mar. Life Sci. Technol. 2021, 3, 363–374. [Google Scholar] [CrossRef]
  26. Wu, Y.; Liu, J.; Liu, D.; Xia, M.; Song, J.; Liang, K.; Li, C.; Zheng, Y.; Wang, M. Microbial metabolic interaction in fermentation ecosystem and cooperation in flavor compounds formation of Chinese cereal vinegar. Food Sci. Hum. Wellness 2024, 13, 3472–3481. [Google Scholar] [CrossRef]
  27. Selegato, D.M.; Castro-Gamboa, I. Enhancing chemical and biological diversity by co-cultivation. Front. Microbiol. 2023, 14, 1117559. [Google Scholar] [CrossRef]
  28. Stanisławska, A.; Szkodo, M.; Staroszczyk, H.; Dawidowska, K.; Kołaczkowska, M.; Siondalski, P. Effect of the ex situ physical and in situ chemical modification of bacterial nanocellulose on mechanical properties in the context of its potential applications in heart valve design. Int. J. Biol. Macromol. 2024, 269, 131951. [Google Scholar] [CrossRef]
  29. Sundaram, M.K.; Nehru, G.; Tadi, S.R.R.; Katsuno, N.; Nishizu, T.; Sivaprakasam, S. Bacterial Cellulose Production by Komagataeibacter hansenii Utilizing Agro-Industrial Residues and Its Application in Coffee Milk Stabilization. Biomass Convers. Biorefin. 2023, 13, 7971–7981. [Google Scholar] [CrossRef]
  30. Hung, C.H.; Cheng, C.H.; Guan, D.W.; Wang, S.T.; Hsu, S.C.; Liang, C.M.; Lin, C.Y. Interactions between Clostridium sp. and other facultative anaerobes in a self-formed granular sludge hydrogen-producing bioreactor. Int. J. Hydrogen Energy 2011, 36, 8704–8711. [Google Scholar] [CrossRef]
  31. Islam, M.U.; Ullah, M.W.; Khan, S.; Shah, N.; Park, J.K. Strategies for Cost-Effective and Enhanced Production of Bacterial Cellulose. Int. J. Biol. Macromol. 2017, 102, 1166–1173. [Google Scholar] [CrossRef]
  32. Wu, S.C.; Li, M.H. Production of Bacterial Cellulose Membranes in a Modified Airlift Bioreactor by Gluconacetobacter xylinus. J. Biosci. Bioeng. 2015, 120, 444–449. [Google Scholar] [CrossRef]
  33. Sharma, C.; Bhardwaj, N.K.; Pathak, P. Rotary Disc Bioreactor-Based Approach for Bacterial Nanocellulose Production Using Gluconacetobacter xylinus NCIM 2526 Strain. Cellulose 2022, 29, 7177–7191. [Google Scholar] [CrossRef]
  34. Pilafidis, S.; Vardaxi, A.; Kourmentza, K.; Pispas, S.; Dimopoulou, M.; Tsouko, E. From Bread Waste to Bacterial Cellulose Nanostructures: Development of a Novel Rotating Disk Bioreactor. Int. J. Biol. Macromol. 2025, 314, 144374. [Google Scholar] [CrossRef]
  35. Lin, D.; Lopez-Sanchez, P.; Li, R.; Li, Z. Production of Bacterial Cellulose by Gluconacetobacter hansenii CGMCC 3917 Using Only Waste Beer Yeast as Nutrient Source. Bioresour. Technol. 2013, 151, 113–119. [Google Scholar] [CrossRef]
  36. Loh, J.; Arnardottir, T.; Gilmour, K.; Muratoglu, M.; Ebrahim, M.; Fernandez, J.; Bennet, P.; Shen, H.; Zhang, L.; Tait, S. Enhanced Production of Bacterial Cellulose with a Mesh Dispenser Vessel-Based Bioreactor. Cellulose 2025, 32, 2209–2226. [Google Scholar] [CrossRef] [PubMed]
  37. Jin, K.; Jin, C.; Wu, Y. Synthetic biology-powered microbial co-culture strategy and application of bacterial cellulose-based composite materials. Carbohydr. Polym. 2022, 283, 119171. [Google Scholar] [CrossRef]
  38. Brugnoli, M.; Mazzini, I.; La China, S.; De Vero, L.; Gullo, M. A microbial co-culturing system for producing cellulose-hyaluronic acid composites. Microorganisms 2023, 11, 1504. [Google Scholar] [CrossRef]
  39. Wang, J.; Tavakoli, J.; Tang, Y. Bacterial Cellulose Production, Properties and Applications with Different Culture Methods—A Review. Carbohydr. Polym. 2019, 219, 63–76. [Google Scholar] [CrossRef]
  40. Zhang, T.Z.; Liu, L.P.; Ye, L.; Li, W.C.; Xin, B.; Xie, Y.Y.; Jia, S.R.; Wang, T.F.; Zhong, C. The production of bacterial cellulose in Gluconacetobacter xylinus regulated by luxR overexpression of quorum sensing system. Appl. Microbiol. Biotechnol. 2021, 105, 7801–7811. [Google Scholar] [CrossRef]
  41. Stephens, K.; Pozo, M.; Tsao, C.Y.; Hauk, P.; Bentley, W.E. Bacterial co-culture with cell signaling translator and growth controller modules for autonomously regulated culture composition. Nat. Commun. 2019, 10, 4129. [Google Scholar] [CrossRef]
  42. Lin, L.; Ma, D.; Wang, Q.; Chen, X.; Zhang, H.; Chen, C. Bottom-up synthetic ecology study of microbial consortia to enhance lignocellulose bioconversion. Biotechnol. Biofuels Bioprod. 2022, 15, 14. [Google Scholar] [CrossRef]
  43. Fei, S.; Yang, X.; Xu, W.; Zhang, J. Insights into proteomics reveal mechanisms of ethanol-enhanced bacterial cellulose biosynthesis by Komagataeibacter nataicola. Fermentation 2023, 9, 575. [Google Scholar] [CrossRef]
  44. Rodrigues, D.M.; da Silva, M.F.; Almeida, F.L.C.; de Mélo, A.H.F.; Forte, M.B.S.; Martín, C.; Barud, H.S.; Baudel, H.M.; Goldbeck, R. A Green Approach to Biomass Residue Valorization: Bacterial Nanocellulose Production from Agro-Industrial Waste. Biocatal. Agric. Biotechnol. 2024, 56, 103036. [Google Scholar] [CrossRef]
  45. Hu, H.; Catchmark, J.M.; Demirci, A. Effects of pullulan additive and co-culture of Aureobasidium pullulans on bacterial cellulose produced by Komagataeibacter hansenii. Bioprocess Biosyst. Eng. 2022, 45, 573–587. [Google Scholar] [CrossRef] [PubMed]
  46. Liu, K.; Catchmark, J.M. Bacterial cellulose/hyaluronic acid nanocomposites production through co-culturing Gluconacetobacter hansenii and Lactococcus lactis under different initial pH values of fermentation media. Cellulose 2020, 27, 2529–2540. [Google Scholar] [CrossRef]
  47. Liu, K.; Catchmark, J.M. Bacterial cellulose/hyaluronic acid nanocomposites production through co-culturing Gluconacetobacter hansenii and Lactococcus lactis in a two-vessel circulating system. Bioresour. Technol. 2019, 290, 121715. [Google Scholar] [CrossRef]
  48. Liu, K.; Catchmark, J.M. Enhanced mechanical properties of bacterial cellulose nanocomposites produced by co-culturing Gluconacetobacter hansenii and Escherichia coli under static conditions. Carbohydr. Polym. 2019, 219, 12–20. [Google Scholar] [CrossRef]
  49. Yu, K.; Chua, S.T.; Smith, A.; Smith, A.G.; Ellis, T.; Vignolini, S. Cultivating future materials: Artificial symbiosis for bulk production of bacterial cellulose composites. bioRxiv 2025. [Google Scholar] [CrossRef]
  50. Li, X.; Chen, Z.; Wang, J.; Mu, J.; Ma, Q.; Lu, X. Symbiosis of acetic acid bacteria and yeast isolated from black tea fungus mimicking the kombucha environment in bacterial cellulose synthesis. Int. Food Res. J. 2023, 30, 1504–1518. [Google Scholar] [CrossRef]
  51. Hidalgo, C.; Vega, R.; García, A.; Romero, J. Metabolic dynamics of the kombucha consortium during fermentation. Int. J. Food Microbiol. 2013, 165, 7–13. [Google Scholar] [CrossRef]
  52. Tu, C.; Li, L.; Wang, Y.; Liu, Y.; Zhou, J.; Liu, X.; Zhu, X.; Zhang, J. Dynamics of Microbial Communities, Flavor, and Physicochemical Properties of Kombucha-Fermented Sargassum fusiforme Beverage during Fermentation. LWT 2024, 192, 115729. [Google Scholar] [CrossRef]
  53. Yuan, Y.; Zhao, B.; Kang, J.; He, C.; Fei, S.; Liu, S.; Qin, X.; Li, C. Lactiplantibacillus plantarum Causes the abnormal fermentation of bacterial cellulose by Komagataeibacter nataicola during nata de coco production. Food Biosci. 2024, 61, 104603. [Google Scholar] [CrossRef]
  54. Fei, S.; Fu, M.; Kang, J.; Luo, J.; Wang, Y.; Jia, J.; Liu, S.; Li, C. Enhancing bacterial cellulose production of Komagataeibacter nataicola through fermented coconut water by Saccharomyces cerevisiae: A metabonomics approach. Curr. Res. Food Sci. 2024, 8, 100761. [Google Scholar] [CrossRef]
  55. Anguluri, K.; La China, S.; Brugnoli, M.; Cassanelli, S.; Gullo, M. Better under stress: Improving bacterial cellulose production by Komagataeibacter xylinus K2G30 (UMCC 2756) using adaptive laboratory evolution. Front. Microbiol. 2022, 13, 994097. [Google Scholar] [CrossRef]
  56. Krystynowicz, A.; Czaja, W.; Wiktorowska-Jeżewska, A.; Gonçalves-Monge, C.; Wronski, T.; Bielecki, S. Factors Affecting the Yield and Properties of Bacterial Cellulose. Pol. J. Environ. Stud. 2002, 11, 467–473. [Google Scholar] [CrossRef] [PubMed]
  57. Ahmadi, T.P.; Nugroho, D.A. Monitoring bacterial cellulose growth during fermentation with various carbon sources by applying real-time image processing. IOP Conf. Ser. Earth Environ. Sci. 2023, 1183, 012031. [Google Scholar] [CrossRef]
  58. Nugroho, D.A.; Sutiarso, L.; Rahayu, E.S.; Masithoh, R.E. Utilizing real-time image processing for monitoring bacterial cellulose formation during fermentation. agriTECH 2020, 40, 118–123. [Google Scholar] [CrossRef]
  59. Moradi, M.; Jacek, P.; Farhangfar, A.; Guimarães, J.T.; Forough, M. The role of genetic manipulation and in situ modifications on production of bacterial nanocellulose: A review. Int. J. Biol. Macromol. 2021, 183, 635–650. [Google Scholar] [CrossRef]
  60. Sayah, I.; Gervasi, C.; Achour, S.; Gervasi, T. Fermentation Techniques and Biotechnological Applications of Modified Bacterial Cellulose: An Up-to-Date Overview. Fermentation 2024, 10, 100. [Google Scholar] [CrossRef]
  61. Behera, B.; Laavanya, D.; Balasubramanian, P. Techno-Economic Feasibility Assessment of Bacterial Cellulose Biofilm Production during the Kombucha Fermentation Process. Bioresour. Technol. 2022, 346, 126659. [Google Scholar] [CrossRef]
  62. Kao, P.M.; Hsu, B.M.; Chang, T.Y.; Chiu, Y.C.; Tsai, S.H.; Huang, Y.L.; Chang, C.M. Biohydrogen production by Clostridium butyricum and Rhodopseudomonas palustris in co-cultures. Int. J. Green Energy 2016, 13, 715–719. [Google Scholar] [CrossRef]
  63. Kang, Y.; Xiao, J.; Ding, R.; Xu, K.; Zhang, T.; Tremblay, P.L. A Two-Stage Process for the Autotrophic and Mixotrophic Conversion of C1 Gases into Bacterial Cellulose. Bioresour. Technol. 2022, 361, 127711. [Google Scholar] [CrossRef] [PubMed]
  64. Prado, M.R.; Blandón, L.M.; Vandenberghe, L.P.S.; Rodrigues, C.; Castro, G.R.; Thomaz-Soccol, V.; Soccol, C.R. Milk kefir: Composition, microbial cultures, biological activities, and related products. Front. Microbiol. 2015, 6, 1177. [Google Scholar] [CrossRef] [PubMed]
  65. Nagaoka, S. Yogurt Production. Methods Mol. Biol. 2019, 1887, 45–54. [Google Scholar] [CrossRef] [PubMed]
  66. Harrison, K.; Curtin, C. Microbial Composition of SCOBY Starter Cultures Used by Commercial Kombucha Brewers in North America. Microorganisms 2021, 9, 1060. [Google Scholar] [CrossRef]
  67. Costa, A.F.S.; Almeida, F.C.G.; Vinhas, G.M.; Sarubbo, L.A. Production of Bacterial Cellulose by Gluconacetobacter hansenii Using Corn Steep Liquor as Nutrient Sources. Front. Microbiol. 2017, 8, 2027. [Google Scholar] [CrossRef]
  68. Yang, Y.; Jia, J.; Xing, J.; Chen, J.; Lu, S. Isolation and Characteristics Analysis of a Novel High Bacterial Cellulose Producing Strain Gluconacetobacter intermedius CIs26. Carbohydr. Polym. 2013, 92, 2012–2017. [Google Scholar] [CrossRef]
  69. Souza, E.F.; Furtado, M.R.; Carvalho, C.W.P.; Freitas-Silva, O.; Gottschalk, L.M.F. Production and Characterization of Gluconacetobacter xylinus Bacterial Cellulose Using Cashew Apple Juice and Soybean Molasses. Int. J. Biol. Macromol. 2020, 146, 285–289. [Google Scholar] [CrossRef]
  70. Liu, Z.; Wang, Y.; Guo, S.; Liu, J.; Zhu, P. Preparation and Characterization of Bacterial Cellulose Synthesized by Kombucha from Vinegar Residue. Int. J. Biol. Macromol. 2024, 258, 128939. [Google Scholar] [CrossRef]
  71. Dourado, F.; Fontão, A.; Leal, M.; Rodrigues, A.C.; Gama, M. Process Modeling and Techno-Economic Evaluation of an Industrial Bacterial Nanocellulose Fermentation Process. In Bacterial Nanocellulose: From Biotechnology to Bio-Economy; Gama, M., Dourado, F., Bielecki, S., Eds.; Elsevier: Amsterdam, The Netherlands, 2016. [Google Scholar] [CrossRef]
  72. Henry, S.; Dhital, S.; Sumer, H.; Butardo, V., Jr. Solid-State Fermentation of Cereal Waste Improves the Bioavailability and Yield of Bacterial Cellulose Production by a Novacetimonas sp. Isolate. Foods 2024, 13, 3052. [Google Scholar] [CrossRef] [PubMed]
  73. Nóvak, I.C.; Segat, B.; Garcia, M.C.F.; Pezzin, A.P.T.; Schneider, A.L.S. Alternative Production of Bacterial Cellulose by Komagataeibacter hansenii and Microbial Consortium. Polímeros 2024, 34, e20240021. [Google Scholar] [CrossRef]
  74. Wang, S.S.; Han, Y.H.; Chen, J.L.; Zhang, D.C.; Shi, X.X.; Ye, Y.X.; Chen, D.L.; Li, M. Insights into bacterial cellulose biosynthesis from different carbon sources and the associated biochemical transformation pathways in Komagataeibacter sp. W1. Polymers 2018, 10, 963. [Google Scholar] [CrossRef] [PubMed]
  75. Ryngajłło, M.; Kubiak, K.; Jędrzejczak-Krzepkowska, M.; Jacek, P.; Bielecki, S. Comparative genomics of the Komagataeibacter genus and insights into bacterial cellulose biosynthesis. Microbiol. Open 2019, 8, e00625. [Google Scholar] [CrossRef]
  76. Liu, H.; Cao, Y.; Guo, J.; Xu, X.; Long, Q.; Song, L.; Xian, M. Study on the isoprene-producing co-culture system of Synechococcus elongatus–Escherichia coli through omics analysis. Microb. Cell Fact. 2021, 20, 6. [Google Scholar] [CrossRef]
  77. Chang, H.; Zheng, Z.; Li, H.; Xu, Y.; Zhen, G.; Zhang, Y.; Ren, X.; Liu, X.; Zhu, D. Multi-omics investigation of high-transglutaminase production mechanisms in Streptomyces mobaraensis and co-culture-enhanced fermentation strategies. Front. Microbiol. 2025, 16, 1525673. [Google Scholar] [CrossRef]
  78. Acharya, D.; Miller, I.; Cui, Y.; Braun, D.R.; Berres, M.E.; Styles, M.J.; Li, L.; Kwan, J.; Rajski, S.R.; Blackwell, H.E.; et al. Omics technologies to understand activation of a biosynthetic gene cluster in Micromonospora sp. WMMB235: Deciphering keyicin biosynthesis. ACS Chem. Biol. 2019, 14, 1260–1270. [Google Scholar] [CrossRef]
  79. Wang, Y.; Chen, Y.; Xin, J.; Chen, X.; Xu, T.; He, J.; Pan, Z.; Zhang, C. Metabolomic profiles of the liquid state fermentation in co-culture of Eurotium amstelodami and Bacillus licheniformis. Front. Microbiol. 2023, 14, 1080743. [Google Scholar] [CrossRef]
  80. Liu, H.; Xian, M.; Cao, Y.; Guo, J.; Kan, L.; Xu, X. Omics integration for in-depth understanding of the low-carbon co-culture platform system of Chlorella vulgaris–Escherichia coli. Algal Res. 2023, 75, 103252. [Google Scholar] [CrossRef]
Applmicrobiol 05 00092 g001
Figure 1. Illustration of BC biosynthesis and nanofiber assembly. Intracellular pathways convert sugars into UDP-glucose, which is polymerized and extruded as cellulose fibers.
Figure 1. Illustration of BC biosynthesis and nanofiber assembly. Intracellular pathways convert sugars into UDP-glucose, which is polymerized and extruded as cellulose fibers.
Applmicrobiol 05 00092 g001
Applmicrobiol 05 00092 g002
Figure 2. Schematic representation of bioreactor configurations for BC production strategies applicable to co-culture systems.
Figure 2. Schematic representation of bioreactor configurations for BC production strategies applicable to co-culture systems.
Applmicrobiol 05 00092 g002
Applmicrobiol 05 00092 g003
Figure 3. Comparative overview of key challenges and future opportunities in co-culture fermentation for BC production.
Figure 3. Comparative overview of key challenges and future opportunities in co-culture fermentation for BC production.
Applmicrobiol 05 00092 g003
Applmicrobiol 05 00092 g004
Figure 4. Schematic representation of omics integration in co-culture fermentation.
Figure 4. Schematic representation of omics integration in co-culture fermentation.
Applmicrobiol 05 00092 g004
Table 1. Progression of co-culture strategies for BC production.
Table 1. Progression of co-culture strategies for BC production.
PhaseConceptual OverviewKey StudiesRef.
Natural ConsortiaTraditional fermented foods such as kombucha and nata de coco relying on undefined microbial communities (yeasts and bacteria) that interact synergistically to produce BC.Kombucha: yeast + Komagataeibacter → ethanol → BC; Nata de coco: K. nataicola + Lactobacillus increased BC production.[16,18]
Defined Dual-Microbe Co-Culture Strategies (2006–2020)Strategic pairing of BC producers with mutualistic partner microbes (e.g., ethanol producers, acid regulators) to enhance and stabilize yields.G. xylinus + L. mali yielded 3× BC yield compared to monoculture; other examples include BC–HA and BC–PHB composite production.[20,38]
Engineered Co-Cultures and Omics-Guided Design (>2020)Integration of synthetic biology, signal-responsive gene regulation, and omics-based optimization for fine control of microbial interactions and BC production.Examples include quorum-sensing-based luxR overexpression in G. xylinus, co-cultures with synthetic signaling modules, and omics-informed pathway engineering in K. nataicola and Komagataeibacter spp.[39,40,41,42,43]
Table 2. Summary of co-culture studies involving Komagataeibacter spp. for enhanced BC production and improvement in material characteristics.
Table 2. Summary of co-culture studies involving Komagataeibacter spp. for enhanced BC production and improvement in material characteristics.
Primary Producer (BC)Co-Culture Partner(s)Fermentation TypeYieldCore Finding (Properties)MechanismRef.
K. xylinusB. cereusCorn stover enzymatic hydrolysateFourfold increase in BC yield with lignocellulosic hydrolysateRobust BC formation under nutrient-rich cultivationAcetoin and 2,3-butanediol from B. cereus enhance BC synthesis[17]
K. nataicola Q2L. fermentumCoconut water-based mediumBC yield increased by up to 59.5%Improved crystallinity, reduced thermal degradation, enhanced water retention and mechanical performanceLAB-derived acids activate Krebs cycle; enhanced β-1,4-glucan formation and cell co-aggregation support fiber assembly[18]
G. xylinusL. maliStatic3-fold increase in BC productionNot analyzedLAB secretes growth-promoting metabolites[20]
K. xylinus MS2530Various yeastsSterilized brewing wasteBC production improved 4–5-fold compared to monoculture; 2–2.5-fold increase with solely brewing wasteIndustrial waste enhances productivity and reduces costYeasts provide ethanol and CO2 that stimulate BC synthesis; waste stream offers nutrients[24]
Komagataeibacter sp.Lactocaseibacillus sp.StaticHigher BC yieldReduced crystallinity, increased fiber size and WHCIn situ incorporation of hyaluronic acid (HA)[38]
K. hanseniiA. pullulansMolasses mediumIncrease of 22.4% over monocultureImproved Young’s modulus and tensile strengthIn situ integration of pullulan into BC matrix[45]
G. hanseniiL. lactisStatic (pH-controlled)Yield varies with pH; best mechanics at pH 4.0Young’s modulus improved to 5029 MPa, altered ribbon width-tuned HA secretion affects BC/HA matrix architectureLower pH showed favorable synergy between G. hansenii and HA secretion from L. lactis[46]
G. hanseniiL. lactis (HA+)Two-vessel circulationEnhanced yield with HA integrationControlled HA production, improved crystallinity and mechanical strengthEngineered HA secretion and stirrer bioreactor to optimize lactic acid production of LAB circulating into BC vessel[47]
G. hansenii ATCC 23769E. coli ATCC 700728StaticYield increased by 10.8% compared to monocultureEnhanced mechanical propertiesIn situ incorporation of mannose-rich exopolysaccharide[48]
K. hanseniiC. reinhardtiiStatic~20% increaseEnhanced 3D BC architecture formation, overcoming oxygen limitationIn situ generation O2 generation by photosynthetic microalgae[49]
K. intermediusB. bruxellensis, Z. bisporusInoculum ratio optimization (1:10:10)Maximum dry weight yield of 5.51 g/L under optimized inoculum ratiosCo-culture promoted efficient substrate conversion and BC assemblyYeasts produced ethanol and growth factors; optimal inoculum ratio also critical for microbes’ interaction[50]
Table 3. Overview of key challenges, representative studies, and mitigation strategies in co-culture-based bacterial cellulose (BC) fermentation.
Table 3. Overview of key challenges, representative studies, and mitigation strategies in co-culture-based bacterial cellulose (BC) fermentation.
AspectKey ChallengesPrior StudiesPotential Mitigation
Strategies		Ref.
Yield of BC and Material CharacteristicsEnhancing BC yield may compromise essential material characteristics such as crystallinity.K. nataicola + L. fermentum SR improved yield but reduced mechanical performance.Real-time monitoring of bacterial cellulose formation combined with genetic modification of strains to tailor material characteristics.[18,19,57,58,59]
Stability and Yield PredictabilityCo-cultures often lack stability, especially under industrial-scale or prolonged fermentations.Pre-fermented coconut water became unstable, especially with more than three species.Designing obligate mutualistic systems and using adaptive laboratory evolution (ALE) to enhance stability.[19,55]
Contamination and Metabolite InterferenceCo-culture partners may produce inhibitory metabolites or alter BC structure.HA-producing LAB enhanced BC yield but reduced crystallinity.Comprehensive chemical and structural characterization to meet regulatory standards for food and biomedical applications.[38,46,47]
Operational Complexity and CostMultispecies cultures require advanced infrastructure and pose economic challenges.Complex systems increase contamination risk and operational costs.Using simplified two-member systems, employing real-time multispecies sensing, and integrating economic modeling.[60,61]
Mutualism vs CompetitionMicrobial interactions may be antagonistic or neutral, not always beneficial.Some strains outcompete others, leading to reduced BC yield.Conducting detailed strain screening and utilizing omics-guided metabolic modeling to select true mutualists.[50,53,54]
Genetic InstabilityKomagataeibacter spp. exhibit adaptive genetic variation in response to environmental stress and consortial conditions.Cellulose synthase operon mutations have been observed, affecting production consistency.Engineering robust Komagataeibacter strains through genomic stabilization and enhanced stress-response regulation.[55,59]
Process ControllabilityIt is difficult to control population dynamics and metabolic flux in real time.Population ratios shift dynamically; substrate pulsing helps but is not universal.Engineering of inducible gene circuits, integrated with feedback control systems utilizing real-time sensor data.[57,58]
Table 4. Comparative techno-economic indicators of BC production. (a) Production cost and key economic metrics for kombucha-based and beet molasses-based systems. (b) Substrate cost comparison. (c) Cost reduction from replacing HS medium with SSF-treated rice bran and cereal dust. Values are extracted from SuperPro Designer® simulations reported by Behera et al. (2022) [61] and Dourado et al. (2016) [71].
Table 4. Comparative techno-economic indicators of BC production. (a) Production cost and key economic metrics for kombucha-based and beet molasses-based systems. (b) Substrate cost comparison. (c) Cost reduction from replacing HS medium with SSF-treated rice bran and cereal dust. Values are extracted from SuperPro Designer® simulations reported by Behera et al. (2022) [61] and Dourado et al. (2016) [71].
(a) Production cost comparison (USD/kg)
ParameterKombucha-based BCBeet molasses-based BC
Capital Investment (USD)13.7213
Operating Cost (USD/year)3.87.4
Production Cost (USD/kg)63.814.8
ROI (%)23.64-
IRR (%)16.48-
Payback Period (years)4.2-
Net Profit (M USD/year)-3.3
(b) Substrate cost comparison (USD/L)
SubstrateCost (USD/L)
HS medium1.36
Black tea + sugar0.1
Enzyme-treated substrate5.58
(c) SSF vs. HS medium cost (AUD/g)
Medium typeCost (AUD/g)
HS medium17.08
SSF-treated RB and cereal dust2.51
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Absharina, D.; Putra, F.J.N.; Ogino, C.; Kocsubé, S.; Veres, C.; Vágvölgyi, C. Bacterial Cellulose Production in Co-Culture Systems: Opportunities, Challenges, and Future Directions. Appl. Microbiol. 2025, 5, 92. https://doi.org/10.3390/applmicrobiol5030092

AMA Style

Absharina D, Putra FJN, Ogino C, Kocsubé S, Veres C, Vágvölgyi C. Bacterial Cellulose Production in Co-Culture Systems: Opportunities, Challenges, and Future Directions. Applied Microbiology. 2025; 5(3):92. https://doi.org/10.3390/applmicrobiol5030092

Chicago/Turabian Style

Absharina, Dheanda, Filemon Jalu Nusantara Putra, Chiaki Ogino, Sándor Kocsubé, Csilla Veres, and Csaba Vágvölgyi. 2025. "Bacterial Cellulose Production in Co-Culture Systems: Opportunities, Challenges, and Future Directions" Applied Microbiology 5, no. 3: 92. https://doi.org/10.3390/applmicrobiol5030092

APA Style

Absharina, D., Putra, F. J. N., Ogino, C., Kocsubé, S., Veres, C., & Vágvölgyi, C. (2025). Bacterial Cellulose Production in Co-Culture Systems: Opportunities, Challenges, and Future Directions. Applied Microbiology, 5(3), 92. https://doi.org/10.3390/applmicrobiol5030092

Article Metrics

Back to TopTop