Advances in Kombucha Tea Fermentation: A Review
- 22 October 2023
- Grondin Eric
- (2)
- Kombucha, Scientific paper
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Citation : Nyhan, L.M.; Lynch, K.M.; Sahin, A.W.; Arendt, E.K. Advances in Kombucha Tea Fermentation: A Review. Appl. Microbiol. 2022, 2, 73-103. https://doi.org/10.3390/applmicrobiol2010005
Copyright : © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Abstract
Kombucha is a carbonated, slightly acidic beverage traditionally produced by the fermentation of sweetened tea by a symbiotic culture of bacteria and yeast (SCOBY). The microbial community of kombucha is a complex one, whose dynamics are still not fully understood; however, the emergence of culture-independent techniques has allowed a more comprehensive insight into kombucha microbiota. In recent times, advancements have been made towards the optimisation of the fermentation process, including the use of alternative substrates, defined starter cultures and the modification of fermentation parameters, with the aim of producing an innovative beverage that is improved in terms of its physiochemical, sensory and bioactive properties. The global kombucha market is rapidly increasing, with the rising popularity of the tea attributed in part to its purported health benefits, despite the lack of research in human subjects to substantiate such claims. Accordingly, the incidence of kombucha home-brewing has increased, meaning there is a requirement for individuals to recognise the potential hazards associated with fermentation and the relevant preventative measures to be undertaken to ensure the safe preparation of kombucha. The aim of this review is to provide an update regarding the current knowledge of kombucha production, microbiology, safety and marketing.
Keywords: kombucha; tea; beverage; fermentation; bacteria; yeast
1. Introduction
2. Kombucha Tea Production
The typical process of kombucha preparation and fermentation is outlined in Figure 1. Under aerobic conditions, kombucha symbiosis can convert sweetened tea into a slightly carbonated, mildly sour and refreshing beverage. The fermentation process essentially exploits the symbiotic relationship between bacteria and yeast. Sucrose is hydrolysed into glucose and fructose by the yeast, resulting in the production of carbon dioxide and ethanol; in turn, LAB utilise glucose and fructose molecules to produce lactic acid, while AAB utilise glucose and ethanol to produce gluconic acid and acetic acid, respectively. Typically, kombucha is prepared by fermenting sweetened black or green with previously fermented liquid tea broth (10–20% v/v) or a tea fungus pellicle. The substrate is incubated statically under aerobic conditions for 7–14 days at 20 °C–30 °C [9].
Figure 1. An overview of kombucha preparation and fermentation.
2.1. Alternative Substrates
Figure 2. Examples of alternative carbon and nitrogen sources used in kombucha fermentation.
2.2. Adjustment of Fermentation Conditions
3. Microbial Diversity
Kombucha fermentation consists of metabolically active bacteria and yeast, which thrive in two mutually non-exclusive compartments: the fermented liquid and the pellicle or biofilm floating on it [7]. The microbial composition of kombucha varies greatly from one batch to another and is dependent on the origin, substrate and fermentation conditions (Table 1). Whilst no yeast has been found to be universally associated with kombucha fermentation, species from genera such as Zygosaccharomyces, Saccharomyces, Brettanomyces, Pichia and Candida are amongst the most common isolates [4,8,61,62,63]. AAB appear to dominate the microbial community, with the main isolated species affiliated with the Komagataeibacter, Acetobacter and Gluconobacter genera. Komagataeibacter xylinus(formerly Gluconacetobacter xylinus) is thought to be one of the most important species associated with kombucha fermentation, due to its superior cellulose-synthesising ability [64,65,66]. LAB are not consistently present; however, the systematic isolation of species from genera such as Lactobacillus, Leuconostoc and Bifidobacterium has been reported [35,67,68].
Table 1. Studies investigating the microbial composition of kombucha.
Use of Defined Starter Cultures
4. Potential Health Benefits
4.1. Clinical Trials
Table 2. Details of controlled human intervention trials examining the effects of kombucha on human health (data correct as of December 2021).
4.2. In Vitro and In Vivo Studies
5. Safety of Kombucha
5.1. Potential Toxicity
5.2. Risk Analysis of Kombucha Production
In terms of chemical hazards, metabolic acidosis is a concern associated with kombucha consumption. Such a condition results from an accumulation of organic acids in the blood, a potentially fatal situation, particularly for those susceptible to acidosis [18]. In Iowa in 1995, two patients presented with severe metabolic acidosis and elevated blood lactic acid levels. Subsequent investigations showed that patient 1 had consumed approximately 4 oz (125 mL) of 7-day-fermented kombucha daily for the previous two months, while patient 2 had recently increased their daily consumption level and fermentation time to 12 oz (375 mL) and 14 days, respectively. The pellicles used by each patient were derived from the same mother pellicle; however, the microbiological analysis showed that pathogenic or toxin-producing microorganisms were not detected, and a causal relationship between kombucha consumption and the patients’ elevated blood acid levels could not be definitively established. [114]. A prolonged fermentation period may result in overproduction of organic acids that pose a possible hazard to the consumer; thus, it is recommended that the fermentation time does not exceed ten days, with a desired final pH of ≥2.5 [18,117]. As is the case with any food or beverage, excess consumption may lead to health problems; thus, kombucha should be consumed in moderation. Alcohol produced during kombucha fermentation can also be classed as a potential chemical hazard, particularly for at-risk populations such as children and pregnant women, for whom even low levels of alcohol consumption can have severe consequences [118,119]. In general, no measures are taken to reduce ethanol production during home brewing of kombucha; however, consumers should be aware of the potentially high alcohol levels in such brews. As commercial kombucha producers often require ethanol contents to be below a specified threshold for the beverage to be classed as non-alcoholic, the fermentation may be stopped after a specified period or the alcohol may be removed post-fermentation. Kombucha is a raw product, and accordingly there is a risk of continued fermentation and subsequent increases in organic acids and ethanol levels if the beverage is not refrigerated. Commercial producers may opt to include a pasteurisation step post-fermentation; however, most choose not to so as to retain the status as a ‘raw’ and accordingly ‘healthier’ product. Chemical contamination of kombucha may also occur if the fermentation is performed in unsuitable containers or vessels. Examples of food-grade containers suitable for kombucha production include those made from glass, stainless steel, high-density polyethylene (HDPE) or propylene (PP), while lead-containing materials such as ceramic or crystal should be avoided due to the risk of leaching toxic constituents from the container. Such an incident has been reported, with a married couple requiring chelation therapy for lead poisoning after daily consumption of kombucha for six months; the patients had brewed the kombucha tea in a ceramic pot, leading clinicians to hypothesise that the acidic tea had leached the heavy metal from the internal pot glazing [120]. However, such an occurrence is rare, with a similar situation not having been reported in the literature since [18].
6. Kombucha Market
6.1. Market Analysis and Current Trends
The global kombucha market stood at USD 1.85 billion in 2019 and is forecasted to increase to USD 10.45 billion by 2027, a projected compound annual growth rate (CAGR) of 23.2%. North America dominated the global market in 2019, with a revenue share of approximately 52%, while Europe is set to hold the second-largest market share in the coming years. In addition, substantial market growth is also being witnessed in Asia–Pacific, the Middle East and Africa [12]. Formed in 2012, the Kombucha Brewers Institution (KBI) is a non-profit trade association of commercial kombucha brewers globally that strives to advance the industry through research, communication and legislation. As of December 2021, 215 commercial kombucha companies across the globe were registered with KBI, with the majority in North America (69.8%), followed by Europe (16.3%) (Figure 3). Unchanged from 2019, the numbers of registered kombucha companies in North America, Latin America, Asia–Pacific and Europe are dominated by Western USA, Mexico, Australia and Spain, respectively. In recent times, there has been increased interest from leading beverage companies seeking to expand their product portfolio and capitalise on the significant growth of the kombucha market. In 2016, PepsiCo acquired California-based company KeVita, a leading fermented beverage company that currently has 4 product lines (Master Brew Kombucha, Sparkling Probiotic Drink, Apple Cider Vinegar Tonic and Prebiotic Shots) spanning 28 flavours [121]. In June 2018, Clearly Kombucha was acquired by Molson Coors Brewing Company, a strategic step to strengthen the company’s non-alcoholic beverage offering [122]. Coca-Cola added its first line of kombucha products in 2018 by acquiring Organic and Raw Trading Co. (producers of MOJO Kombucha), adding to its ever-increasing Australian beverage portfolio of 165 products and 25 brands at the time [123]. Coca-Cola’s continued interest in the kombucha market resulted in a $20 million equity investment in Health-Ade Kombucha in 2019 [124], a company that later sold a controlling stake to longstanding partner First Bev [125]. In 2018, Peet’s Coffee (a USA company producing ready-to-drink (RDT) coffee) acquired a majority stake in Revive Kombucha for an undisclosed amount [126].
6.1. Market Analysis and Current Trends
Figure 3. Number of Kombucha Brewers International (KBI) registered companies and their respective regions. Data correct as of December 2021.
6.2. Commercial Kombucha Products
Table 4. Examples of commercial kombucha products on the market.
7. Conclusions and Future Perspectives
The global popularity of kombucha is rising significantly, with increases in both household and commercial production of the fermented tea. The beverage’s microbiological, sensory, and functional compounds vary and are highly dependent on the substrate used, the fermentation conditions and the microbial diversity of the kombucha consortium. Sucrose and black or green tea are the primary fermentation substrates; however, the use of alternatives has been investigated. Different substrates can positively or negatively influence the kombucha beverage, affecting the microbial growth, metabolite production and functionality. Yeasts and AAB are the core microbiota of the kombucha community, while LAB have also been identified systematically. Microbial diversity at genus and species levels can vary between consortiums in different geographical regions and even within the same region. Yeasts play a key role in making substrates available to AAB through a mutualistic relationship, while AAB utilises such substrates to produce organic acids and other metabolites, which positively modify the environment for yeast. The emergence of culture-independent techniques has allowed for a deeper insight into the diversity of the complex microbial community and the interactions taking place therein. The use of defined starter cultures to reproduce a stable kombucha community is a promising alternative to traditional kombucha fermentation; however, this area of research remains largely understudied. The safety of kombucha is typically guaranteed by its low pH; however, producers and consumers should still be aware of the risks, such as biological contamination and secondary fermentation. There is a widespread belief that kombucha is a health product; however, its purported benefits have been investigated using animal models and human cell lines, while little in the way of research on kombucha’s function in human subjects has been performed to date. Future studies should focus on the modulation of kombucha’s sensory and functional characteristics through a deeper understanding of the communication and interactions between members of the microbial community. In addition, research efforts should be directed towards the use of defined starter cultures, particularly to facilitate a shift toward industrial-scale production of kombucha.
Conceptualisation, E.K.A.; investigation, L.M.N.; writing—original draft preparation, L.M.N. and K.M.L.; writing—review and editing, L.M.N., K.M.L., A.W.S. and E.K.A.; supervision, E.K.A. All authors have read and agreed to the published version of the manuscript.
This research received no external funding.
Not applicable.
This work has been sponsored by and performed in collaboration with AB-InBev’s GITeC—The Core Global Research and Development Centre. The authors would like to thank Stuart Wilkinson, Luk Daenan and Patrick O’ Riordan for their support of this work. The authors would also like to thank William Organ and Audrey Byrne for their assistance throughout the writing of this manuscript.
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