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Could non-Saccharomyces yeasts contribute on innovative brewing fermenta-
tions?

Rafael Felipe Basso, André Ricardo Alcarde, Cauré Barbosa Portugal

PII: S0963-9969(16)30233-2
DOI: doi: 10.1016/j.foodres.2016.06.002
Reference: FRIN 6297

To appear in: Food Research International

Received date: 24 April 2016


Revised date: 3 June 2016
Accepted date: 4 June 2016

Please cite this article as: Basso, R.F., Alcarde, A.R. & Portugal, C.B., Could non-
Saccharomyces yeasts contribute on innovative brewing fermentations?, Food Research
International (2016), doi: 10.1016/j.foodres.2016.06.002

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Could non-Saccharomyces yeasts contribute on innovative brewing fermentations?

Rafael Felipe Bassoa, André Ricardo Alcardea, Cauré Barbosa Portugala

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Dept. Agroindústria, Alimentos e Nutrição, Escola Superior de Agricultura “Luiz de Queiroz” -

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University of São Paulo, Avenida Pádua Dias 11, 13418-900, Piracicaba, Brazil.

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Corresponding author: Cauré Barbosa Portugal

Email: caure.portugal@gmail.com. Telephone +55 19 982644658

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Abstract
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With the advances in the production of beer worldwide, more challenges arise each year in the search

for new approaches to the development of distinctive beverages. Attempts to obtain products with
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more complex sensory characteristics have led experts and brewers to prospect for non-conventional
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yeasts, i.e., non-Saccharomyces yeasts that may provide a new range of perspectives in terms of

techniques and approaches. Besides the widespread use of Dekkera/Brettanomyces for the production
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of sour beers, other species are emerging for presenting unusual metabolic features that include the
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production of fruity esters, and a distinctive enzymatic apparatus. Wickerhamomyces anomalus and

Torulaspora delbrueckii stand out as the most promising yeasts in brewing processes. Such new

tendencies in the use of non-Saccharomyces yeasts comprise the production of low-alcohol beers,

functional beers, and bioflavoring approaches. This is a little explored field in brewing practice, still

requiring extensive research with practical application. In this sense, this review aims to present the

main points for the application of non-conventional yeasts in beer production, and thus contribute to

future advances in the topic.

Keywords: non-conventional yeasts; brewing; specialty beers; low-alcohol beers; biotechnology;

bioflavoring
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Chemical compounds

Chemical compounds studied in this article

Ethanol (PubChem CID: 702); Acetic acid (PubChem CID: 176); Glucose (PubChem CID: 5793);

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Maltose (PubChem CID: 6255); Maltotriose (PubChem CID: 192826); Ethyl acetate (PubChem CID:

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8857); Isoamyl acetate (PubChem CID: 31276); Ethylguaiacol (PubChem CID: 62465); Ethylphenol

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(PubChem CID: 31242)

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Acknowledgements: This work was supported by Sao Paulo Research Foundation (FAPESP) [grant

number 2013/03766-4].

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1. Introduction

The brewing of beer-like beverages already occurred between 5500-3100 BC in the regions of

Mesopotamia and Ancient Egypt (Hornsey, 2007). Barley grew in wild forms in these regions, and

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such civilizations found that moistening, germinating, and drying the grains would lead to a sweeter,

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less perishable product (Papazian, 2003). This is what we now perceive as barley malt.

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Archaeological evidences suggest that in such times beer was made from barley, wheat, or a mixture

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of both. The process was carried out by spontaneous fermentations, either by preparing a cereal

slurry, or by using a mixture of raw grains, cooked malt and water, with further exposure to ambient

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air to induce “contamination” (Sicard & Legras, 2011).
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Since the species of the genus Saccharomyces – especially S. cerevisiae – frequently appeared as

abundant and dominant agents in spontaneous fermentations, they were intuitively selected over

generations, in different cultures and from distinct raw materials. Readily, men figured out that using
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a small piece or volume of a modified product into a fresh one, it worked as a starter for inducing
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fermentation again. It was only in the mid 17th century scientists began to understand what was

behind fermentation, enabling the isolation techniques, selection and use of pure inocula in those
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processes (Steensels & Verstrepen, 2014). Successive investigations, that included yeast’s
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description by Antonie van Leeuwenhoek in 1680, its association with beer wort fermentation by

Cagniard de Latour in 1836, the establishment of isolation techniques, and the use of pure starters as

top or bottom fermenting yeasts by Emil Hansen in 1883, enabled the improvement of controlled and

industrial processes (Lodolo, Kock, Axcell, & Brooks, 2008).

There are many different styles of beer but, primarily, the main brewing classification criterion

particularly relies on the type of fermentation. Although this concept may vary, brewing yeast are

mainly classified in top-fermenting or ale yeasts (Saccharomyces cerevisiae), and bottom-fermenting

or lager yeasts (Saccharomyces pastorianus). The two main styles of beer are still based on such

traits, of which top-fermenting yeasts display more genetic divergence, and the process normally
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occurs between 18 ºC – 24 ºC, while bottom-fermenting yeasts show a conserved genetic material,

and often ferment at lower temperatures (8 ° C – 14 ° C) (Lodolo et al., 2008). Fermentations for

Lambic beers, on the contrary, is a spontaneous, uncontrolled process, driven by brewery-resident

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microorganisms that are introduced by exposing the wort in shallow tanks during the overnight

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cooling, before transferring it to wooden barrels for fermentation and aging.

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Since the establishment of sedentary life by mankind, and formation of the first human social

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structures, multiple events – including natural and human selection – led to a pool of domesticated or

selected yeasts (Sicard & Legras, 2011). Selected Saccharomyces strains are used with various

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purposes because of their plasticity on assimilating different substrates, usually not incorporated by
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S. cerevisiae. The use of Saccharomyces strains in controlled fermentations over decades is

essentially based on three main features: (a) efficient production of high ethanol amounts; (b) the use

of fermentation as the preferential metabolic pathway, combined to the positive Crabtree effect
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(repression of respiration by glucose); and (c) higher tolerance to ethanol and other environmental
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stresses (Steensels & Verstrepen, 2014).

Nonetheless, with increasing demands for innovative products, other non-Saccharomyces species
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have been isolated and characterized for the development of specialty beers, with distinctive
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aromatic and flavor components (Vanderhaegen et al., 2003). They generally present low

fermentation yields, and are more sensitive to ethanol stress (Di Maro, Ercolini, & Coppola, 2007),

but may display a great range of possibilities for beer fermentations in order to provide distinctive

aroma and flavor (Renouf & Lonvaud-Funel, 2007). It has been shown, for example, that Dekkera

bruxellensis and Hanseniaspora uvarum (anamorph, Kloeckera apiculata) have the ability to

produce many esters with fruity character, besides promoting the enzymatic breakdown of some

proteins (De Keersmaecker, 1996; Kumara & Verachtert, 1991). Lately, other wild yeast species

have been mentioned as interesting alternatives in brewing by their genome particularities, as in the

case of D. anomala, Hanseniaspora uvarum, S. bayanus (= S. uvarum), S. pastorianus,


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Naumovozyma dairenensis (= Naumovia dairenensis), and Debaryomyces spp. These

microorganisms can display unusual metabolic abilities, thus changing the sensory character of the

final product by adding complexity in flavor, and modifying mouthfeel sensation (Spitaels et al.,

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2014). Some yeasts are eventually characterized as spoilage agents, as in the case of

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Dekkera/Brettanomyces and Debaryomyces – especially in poorly controlled processes in which they

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unintentionally appear. However, many of them are already used as pure inoculum, or in yeast blends

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under controlled conditions of processing for specialty beer production (Hayashi, Arai, Tada,

Taguchi, & Ogawa, 2007).

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In most beers, the microorganisms that carry out fermentation belong to different Saccharomyces
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species and strains (Bokulich & Bamforth, 2013). The occurrence of other species is commonly

reported in some peculiar beer styles produced by spontaneous fermentations, as the Belgian

Lambics and Gueuzes, and their American counterparts, Coolship Ales. However, due to the
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diversity of substrate assimilation patterns that non-conventional yeasts may display (van Dijken,
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2002), the use of non-Saccharomyces in controlled fermentations has been gaining popularity among

brewers in order to obtain distinctive products, a key point to gain market share in this growing
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segment (Ciani & Comitini, 2011; Cordero-Bueso, Esteve-Zarzoso, Cabellos, Gil-Díaz, & Arroyo,
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2013; González, Quirós, & Morales, 2013; Johnson, 2013). To take one example, the market of craft

beers in the United States grew 12.8 % in 2015, increasing 16.3 % in exports, and with incomes of

22.3 billion dollars, opening 620 craft breweries – only 68 closings in the same period – and

establishing a goal of achieving 20 % of beer market share by 2020 (Brewers-Association, 2016;

Kell, 2016). A significant strengthening of the sector has also been detected in several European

countries, where growth sometimes outstripped 300-1000% from 2009 to 2014 (Table 1).

The production of quality beer relies on the activity of fermenting yeasts that are qualified not only

for good fermentation yield-efficiency, but also for printing aroma and flavor to the beverage. Their

importance in beer production goes beyond the formation of ethanol and carbon dioxide, since most
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of the compounds that provide taste and aroma arise during fermentation, and these are intermediate

compounds and by-products of metabolism (Pires, Teixeira, Brányik, & Vicente, 2014).

In this sense, this review aims to recover and discuss recent information on the status of non-

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conventional yeasts, and their potential application in beer production. Even so, this work intends to

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present the main technological approaches regarding the use of non-Saccharomyces as a way to

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improve sensory traits, as well as product diversification and innovation in brewing processes.

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2. Non-Saccharomyces yeasts in brewing

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Saccharomyces cerevisiae is the widespread brewing yeast, and selected strains have proportionated

improvement and control of processes as a way to obtain chemical and sensory quality, as well as to
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maintain reproducibility. The expansion in the sector, and the growing number of specialized

consumers, have driven brewers to innovate methods changing hop varieties, using special malts,
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preparing distinctive blends, and more. Nevertheless, it is in the soul’s beverage, in the fermentation
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and pitching non-conventional yeasts, where producers are finding not only a multiplicity of aroma

and flavor production, but also new approaches and impressions in the overall organoleptic profile of
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beers (Michel et al., 2016). In this sense, some old acquaintances and other promissory
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microorganisms have outlined an ever-increasing range of techniques and sensory results in brewing.

In Table 2 we summarize the main yeast species discussed in this work, presenting their current

taxonomical status, major isolation sources recorded in literature, as well as their food safety

classification. Additionally, Table 3 brings most prominent information about their major aromatic

compounds produced and descriptors, in addition to the related enzymes and peculiar traits of each

species. In the following topics, we discuss their specific characteristics, and thereafter, some

technology and product innovation regarding the use of these non-conventional yeasts.
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2.1 Dekkera/Brettanomyces

In the early 1970s, almost all known species of Dekkera/Brettanomyces had been isolated from

spontaneous brewing processes. Today, they are the most relevant non-conventional yeasts in the

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production of sour beers, mainly by the discovery of its singular influence in Lambic beers, and have

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attracted growing interest from brewers and connoisseurs. Lambics are produced under a complex

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succession of spontaneous fermentations, in which Dekkera/Brettanomyces yeasts take part during

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the acidification-maturation stage, a process occurring after three or four months after fermentation

starts (Spitaels et al., 2014). This final maturation phase may last up to ten months, and it is known

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that these yeasts are responsible for producing a set of flavor compounds, mainly ethyl phenol, ethyl
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guaiacol, isovaleric acid, acetic acid, and ethyl acetate, which together result in a typical flavor

character (Vanderhaegen et al., 2003; Verachtert & Derdelinckx, 2014). However, an increasing

number of researches addressing to the use of these yeasts in controlled fermentations has been
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observed in the last years, both in pure cultures as in co-inoculation with S. cerevisiae (Steensels &
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Verstrepen, 2014).

Beyond their typical phenolic notes, those commercial Dekkera/Brettanomyces strains have shown
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the ability to exert a marked influence on the sensory profile, contributing to an overall fruity or
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floral character by producing high levels of ethyl esters (ethyl acetate, ethyl caprate, ethyl caprylate,

ethyl lactate) (Crauwels et al., 2015). It is noteworthy that some peculiar flavors – namely “horse

blanket”, “stable” character – may also arise, and in small concentration, account for a positive and

complex bouquet. On the other hand, misguided fermentations with numerous

Dekkera/Brettanomyces communities may lead to some unpleasant off-flavors that include “acrid

smoke”, “band-aid” and “fecal” descriptors. The unique taste and aroma traits they are able to

imprint is a fact particularly related to the activity of certain amylases, although it a strain-dependent

trait (Daenen, Sterckx, Delvaux, Verachtert, & Derdelinckx, 2008).


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Since its first description in 1904 by Niels Hjelte Claussen, director of the Carlsberg brewery at that

time, there were several taxonomic changes for the genus. Currently, Dekkera is the official

nomenclature, adopted for the sexual or teleomorph form, and Brettanomyces designates the

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anamorphic or asexual form of this yeast (Kurtzman, 2011). D. bruxellensis and D. anomala are the

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species that typically appear in brewing processes (Steensels et al., 2015; White & Zainasheff, 2010).

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Besides presenting certain characteristics that enable their good performance in alcoholic

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fermentations, including tolerance to osmotic pressure, ethanol, limiting oxygen and low pH, these

species show a diverse metabolism that leads to interesting and unusual results in beer production

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(Steensels et al., 2015).
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When compared to S. cerevisiae, these yeasts have greater genetic diversity, and may present higher

gene doses due to increased ploidy variability, especially in low-oxygen and high sugar

concentration environments (Piškur et al., 2012; Woolfit, Rozpędowska, Piškur, & Wolfe, 2007). D.
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bruxellensis and D. anomala usually display the Crabtree effect, promoting fermentation instead of
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respiration, even with available oxygen in media with a certain concentration of sugars (Thomson et

al., 2005). However, the metabolic trait that markedly differentiates them from those traditional
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yeasts is the expression of the Custer effect. In this case, these organisms have greater ability to
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conduct the fermentation in the presence of oxygen by reducing or ceasing the glucose metabolism

(lag phase) under anaerobic conditions (Barnett & Entian, 2005). It is worth mentioning that small

oxygen inputs during fermentation is encouraging to best results in biomass production, fermentation

efficiency, total consumption of glucose, ethanol production, and low production of acetic acid

(Aguilar-Uscanga, Délia, & Strehaiano, 2003).

Dekkera/Brettanomyces yeasts are able to metabolize a wide range of carbon sources, including

cellobiose and dextrins. These sugars are not readily assimilated by Saccharomyces species, except

in the case of S. cerevisiae var. diastaticus, which is able to assimilate dextrin and starch (Steensels

et al., 2015). Cellobiose is a disaccharide found in wooden barrels used to mature beers (Daenen,
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Sterckx, et al., 2008; Vanderhaegen et al., 2003), and dextrins are more complex sugars that include

maltotetraose and maltopentaose. Such molecules are the main components of the residual sugar

content in beers fermented by S. cerevisiae, and can be degraded by the β-glucosidase. This enzyme,

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commonly produced by Dekkera/Brettanomyces, enables degradation of those complex molecules

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into simple sugars that can be readily assimilated, resulting in a "super-attenuated" beer. The

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aforementioned methodology includes some particular traits, and practical results in beer styles in

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which low calorie and residual sugar content constitute innovative hot spots (Kumara, De Cort, &

Verachtert, 1993; Steensels et al., 2015).

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Regarding the influence of temperature on fermentations driven by Dekkera/Brettanomyces, most
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studies focus on biomass production rate, primary products (ethanol, acetic acid and carbon dioxide),

and glucose consumption patterns. Fermentations processed between 20 ºC – 32 ºC result in better

ethanol production, as well as lower acetic acid contents released in the medium (Brandam, Castro-
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Martínez, Délia, Ramón-Portugal, & Strehaiano, 2007). High concentration of these compounds can
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directly affect cell viability, aptitude in bioconversion of substrates, and low fermentative capacity.

Besides disturbing yeast growth and leading to acidic shock conditions, intense release of acetic acid
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also increases the risk of impairing the volatile acidity and contribute to sensory imbalance in the
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final product (Rogers, Veatch, Covey, Staton, & Bochman, 2016).

Although the tolerance to ethanol is a strain-dependent factor, D. bruxellensis is noticeably resistant

to concentrations of up to 13.5 – 15.0 % (v/v) (Barata et al., 2008), although in vitro tests show that

some strains can tolerate up to 20 % ethanol (Portugal et al., 2014). D. anomala, otherwise, is more

sensitive to its toxicity, tolerating at most 8 % (v/v) ethanol in the medium (Barata et al., 2008). In

any case, the response of these microorganisms to ethanol stress entails on the extension of the lag-

phase, with direct consequences on growth rate and cell density, in addition to increased

concentrations of ethyl phenols and ethyl esters in the medium (Conterno, Aprea, Franceschi, Viola,

& Vrhovsek, 2013).


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The production of acetic acid by Dekkera/Brettanomyces is part of the strategy based on “make-

accumulate-consume”. Such system involves an interesting evolutionary feature that gives it a

competitive edge, characterized by the production and accumulation of ethanol and acetic acid in the

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fermentative environment, with subsequent degradation of these compounds when fermentable

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sugars are depleted (Rozpędowska et al., 2011). Although considered a fault in many beer styles, low

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concentrations of acetic acid (< 1.2 g/l) are acceptable in those ones that use Dekkera/Brettanomyces

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in their primary fermentation (Steensels et al., 2015). Higher concentrations of this compound,

however, appear to inhibit substrate consumption. The metabolic regulation of this by-product is

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highly dependent on oxygen availability, with a direct relation between oxygen concentration in the
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substrate and the acetic acid production (Aguilar-Uscanga et al., 2003). It is known, moreover, that

the temperature influences only slightly in the production of this compound (Brandam et al., 2007).

There is a set of very peculiar sensory characteristics that Dekkera/Brettanomyces imparts to the
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beverage, mainly by the production of phenolic off-flavors (POF) that are frequently described as
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“rubber”, “burnt plastic”, “medicinal”, “leather”, and “barnyard”. Much of this influence relies on

the ability to metabolize hydroxycinnamic acids, of which ferulic, p-coumaric and caffeic acids are
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the most common (Chatonnet, Dubourdie, Boidron, & Pons, 1992; Lentz & Harris, 2015). These
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compounds are especially present in the bark, pericarp, aleurone, and endosperm of cereal grains,

associated to the arabinoxylans structural carbohydrates, and incorporated to wort during the

mashing process (Vanbeneden, Van Roey, Willems, Delvaux, & Delvaux, 2008; Yu, Vasanthan, &

Temelli, 2001). Dekkera/Brettanomyces has two enzymes that work to reduce such acids into ethyl

phenols in two consecutive main reactions: a) the first one takes place by the action of the enzyme

cinnamate decarboxylase, which metabolizes the acids in their respective vinyl phenols; b) the

second enzyme is the vinylphenol reductase, which reduces those vinyl phenols into ethyl phenols.

(Harris, Ford, Jiranek, & Grbin, 2009; Vanbeneden, Gils, Delvaux, & Delvaux, 2008). The presence

of these last compounds above the perception threshold is considered undesirable in most beer styles;
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however, they are essential in many others, including the Belgian Lambic, Gueuze and Red Flanders,

the German Weizen and Rauchbier, the American Coolship Ale, as well as the Sour and Farmhouse

Ales (Lentz & Harris, 2015).

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Primary fermentations using Dekkera/Brettanomyces generally lead to beers with low content of

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isoamyl acetate and ethyl phenylethyl, but instead, high concentrations of ethyl acetate and ethyl

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lactate (Spaepen, Van Oevelen, & Verachtert, 1978; Spaepen & Verachtert, 1982). Moderate levels

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of these latter compounds are desirable, and may contribute to the flavor complexity of beers, but

higher concentrations may easily impart undesirable solvent-like, buttery and medicinal notes to the

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beverage (Cortés-Diéguez, Rodriguez-Solana, Domínguez, & Díaz, 2015). Ester imbalance occurs
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with the degradation of acetate esters by the esterase enzyme, a bioprocess much more efficient than

the hydrolysis of non-acetate esters (Steensels et al., 2015).

Another striking feature on Dekkera/Brettanomyces is the production of β-glucosidase, an enzyme


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responsible for the hydrolysis of glycosides. Glycosides are non-volatile compounds composed by
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one aromatic molecule (aglycone) and one β-D-glucose (glycone), with possible additional sugar

units attached (Winterhalter & Skouroumounis, 1997). Such compounds are commonly found in
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plants, flowers and fruits, and some of them are used in brewing, and are present in hops and in the
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wood of barrels employed for maturation (Daenen, Saison, et al., 2008). The enzymatic hydrolysis of

glycosides releases the sugar molecules, further metabolized, and the aglycone, which can express

aromatic activity, representing a poorly explored methodology to improve the aromatic potential of

beers (Winterhalter & Skouroumounis, 1997). The β-glucosidase activity is a strain-dependent

feature, but D. bruxellensis stands out for presenting such hydrolytic action on a number of

substrates, emerging as a good option for fermentations in pure culture or in co-culture with S.

cerevisiae (Daenen, Saison, et al., 2008; Daenen, Sterckx, et al., 2008).


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2.2 Other yeasts

Although Dekkera/Brettanomyces is currently the most prominent yeast in distinctive approaches,

other genera and species are reported to occur in brewing processes. Wickerhamomyces anomalus

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and Torulaspora delbrueckii currently stand out as the most relevant non-conventional yeasts for

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brewing due to their flavoring features, tolerance to extreme environmental conditions, low

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production of undesirable by-products, and frequent association with food and beverages (Canonico,

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Agarbati, Comitini, & Ciani, 2016; Michel et al., 2016; Schneider et al., 2012). Nonetheless, other

yeasts of the genera Wickerhamomyces, Candida, Torulaspora, Hanseniaspora, and Debaryomyces,

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usually occurring and isolated from spontaneous fermentations, have also been investigated. The
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most common styles in that cases include the Belgian Lambics and the American Coolship Ale, in

which several yeast species contribute to the typical, particular characteristics of the beer (Bokulich

& Bamforth, 2013; Spitaels et al., 2014).


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2.2.1. Wickerhamomyces anomalus


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In contrast to S. cerevisiae and Dekkera/Brettanomyces, Wickerhamomyces anomalus (= Pichia

anomala) is able to regulate the central metabolism of carbon according to oxygen availability
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instead of doing so by the glucose concentration (Crabtree-negative), increasing the glycolytic


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pathway under oxygen restriction (Pasteur effect). It enables the fermentative pathway in limited

oxygen conditions, in which the enzymes pyruvate decarboxylase and alcohol dehydrogenase are

activated, but rather small concentrations of ethanol are produced in aerobic conditions (Fredlund,

2004; Passoth, Fredlund, Druvefors, & Schnürer, 2006). W. anomalus shows a wide spectrum of

sugar assimilation, being able to properly metabolize hexoses (glucose, galactose, fructose), pentoses

(arabinose, xylose), disaccharides (sucrose, lactose), polysaccharides, as well as some alcohols,

organic acids, fatty acids, and aromatic hydrocarbons (Walker, 2011). This species also displays

great physiological plasticity, being able to grow in absence of oxygen, in wide temperature range

(between 3 ºC and 37 ºC), and under extreme pH values between 2.0 – 12.4 (Fredlund, 2004). Strains
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of this genus are somewhat sensitive to ethanol and acetate (Passoth et al., 2006), but show tolerance

to high osmotic pressure, and are efficient in their mechanisms of adaptation to stress factors

(Walker, 2011).

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Regarding its contribution to sensory active compounds, this species is a good producer of ethyl

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propanoate, phenyl ethanol, 2-phenylethyl acetate, and especially ethyl acetate (Passoth et al., 2006).

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In contrast to S. cerevisiae, that synthesizes ethyl acetate mainly from acetyl-CoA and ethanol by the

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action of alcohol acetyltransferase (Yoshioka & Hashimoto, 1981), W. anomalus makes it mainly by

reverse reaction of esterase from acetate and ethanol (Rojas et al., 2002; Yoshioka & Hashimoto,

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1981). Such compound is metabolically important because of its antifungal activity (Fredlund,
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Druvefors, Olstorpe, Passoth, & Schnürer, 2004), besides the property of preventing toxic

accumulation of acetic acid into the cells in limited oxygen conditions (Fredlund, 2004). From a

sensory standpoint, ethyl acetate directly influences on the taste and flavor profile of the beer,
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lending an interesting fruity or an unpleasant solvent-like character, depending on the concentration


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values (White & Zainasheff, 2010).

Despite some evidences suggest an apparent inability to metabolize maltose (Walker, 2011), other
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studies show that some wild W. anomalus strains are indeed able to use this sugar and to grow even
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better than other commercial brewing yeasts (Y.-J. Lee et al., 2011). W. anomalus stands as a good

candidate in sequential or co-inoculated fermentations with other yeasts, always considering its

inhibitory action against some microorganisms (Fredlund et al., 2004; Passoth et al., 2006; Walker,

2011).

2.2.2. Torulaspora delbrueckii

Torulaspora delbrueckii (anamorph Candida colliculosa) is a species whose use supposedly dates

back over 4000 years, and it is well studied today in wine controlled fermentation processes because

of its good production of fruity flavors (Albertin et al., 2014). Some authors assume that T.

delbrueckii is also one of the main responsible by the fermentation of Bavarian wheat beers (Michel
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et al., 2016; Tataridis, Kanelis, Logotetis, & Nerancis, 2013). These microorganisms can undergo

high osmotic pressure conditions (highly osmotolerant) (Alves-Araújo et al., 2007; Bely, Stoeckle,

Masneuf-Pomarède, & Dubourdieu, 2008), grow well at low temperatures (cryotolerants), and

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curiously require oxygen to ferment – although not displaying the Custer effect (Alves-Araújo et al.,

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2007; Hanl, Sommer, & Arneborg, 2004; Salvadó et al., 2011).

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T. delbrueckii shows invertase activity, necessary for sucrose hydrolysis, and subsequent

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consumption of glucose and fructose. The ability to consume maltose – main sugar in wort – is

conditioned by the activity of the enzyme maltase, as well as by maltose transporters also present in

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this species, highlighting that such biochemical apparatus can be inhibited by high concentrations of
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glucose in the medium. Interestingly, the ability to metabolize the sugar maltotriose was also proven

in this microorganism. The rate and extent to which the different sugars are consumed depend mainly

on the strain, with some of them being unable to consume maltose and maltotriose (Alves-Araújo et
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al., 2007; Michel et al., 2016).


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Ethanol tolerance and fermentation efficiency are also strain-dependent features. Although it has

been reported that T. delbrueckii is able to tolerate up to 11 % (v/v) ethanol in winemaking


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conditions (Tataridis et al., 2013), recent evidences demonstrate that many strains are only able to
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grow in concentrate beer wort with at most 5 % ethanol in the medium (Michel et al., 2016).

Tolerance to iso-α-acids from hops is another important trait that yeast should display in brewing

conditions, once that compound has inhibitory effect against some microorganisms, including gram-

positive bacteria and yeasts. T. delbrueckii can grow – even with an increase in the lag-phase – in the

presence of up to 90 ppm iso-α-acids in the medium, a concentration that correlates to highly hopped

beer styles. Resistance to such compounds derived from hops can be evident even in the presence of

alcohol, since T. delbrueckii is able to grow in wort with an ethanol content of up to 5 % (v/v), and

can still manage levels of up to 50 mg/l iso-α-acid (Michel et al., 2016). Of course, these are strain-

dependent traits, but the ability to deal with some ethanol content in the medium – toxic to the cell –
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provides better membrane selectivity conditions. As a result, some strains of T. delbrueckii are less

sensitive to antimicrobial effects of iso-α-acids, and may be proposed as a tool for average strength

and more hoppy beers.

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The most interesting contribution of this yeast is perhaps the profile of volatile compounds it may

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confer to the final beer. Some recent investigations reveal the ability of some strains to produce 2-

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phenylethanol (sweetish, “rose petals” and floral flavors) (Etschmann et al., 2015), n-propanol and i-

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butanol, amyl alcohol (“solvent brandy” aroma) (Pires et al., 2014), and ethyl acetate (Meilgaard,

1982). T. delbrueckii seems to produce low amounts of isoamyl acetate (Hernández-Orte et al.,

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2008), as in the case of ethyl hexanoate, ethyl octanoate and ethyl decanoate, whose concentrations
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were negligible, and below the perception threshold (0.01 mg/l) (Michel et al., 2016). Unlike

Dekkera/Brettanomyces, the enzymatic apparatus necessary to hydroxycinnamic acids’ reduction is

not present in T. delbrueckiii, so that this species does not have the inconvenience of producing
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phenolic off-flavors. The same is not true in the case of diacetyl production, since it often releases
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high amounts of this compound, which cannot be reduced during bottle or cask conditioning. Taken

as a whole, the beverages brewed with T. delbrueckiii show honey and pear-like flavors, with highly
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citric and fruity character on the palate (Michel et al., 2016).


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3. Technology and product innovation

The specialty beer market has gained attention worldwide in the last 30 years. In addition to the

evidence of beers produced with better quality inputs, and using methods that respect the final

product attributes, the “liquid bread” is gaining other perspectives. These new insights include the

low-calorie beers, low alcohol or alcohol-free beers, gluten-free beers, functional beers, and brewing

processes that would enable the incorporation of new flavors to the beverage (Yeo & Liu, 2014). As

well as in winemaking, brewing technology and product innovation are greatly motivated by

exploring specific traits of new yeast species. Display of peculiar and interesting by-products or

secretion of extracellular enzymes, besides singular fermentative ability or sugar assimilation traits,
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integrate some of the tools available for improving brewing methods and results (Masneuf-

Pomarede, Bely, Marullo, & Albertin, 2015). Some of these approaches simply rely on recuperating

traditional practices, but new perspectives look at bioconversion processes to enhance flavor

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characteristics, to obtain products with low calorie and/or alcohol contents, and even being able to

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provide beers with current complexions of a functional food.

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3.1. Recovering old production methods for innovation

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The one-process fermentation-maturation-aging in wooden barrels was a widely used methodology

in the past, probably by the lack of choice. It is no longer extensively applied due to the growing

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popularity of pure yeast cultures and stainless steel vats. Today, this procedure has been gaining
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interest especially in craft breweries, representing a way of differentiation and innovation of product.

This practice renders the micro-oxygenation of beer, besides contributing with tannins, body and

flavor complexity (Tonsmeire, 2014). Another interesting feature of barrels is that wood actually
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harbor some wild yeasts, lactic acid bacteria, and acetic acid bacteria (Bokulich, Bamforth, & Mills,
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2012; Verachtert & Derdelinckx, 2014). These microorganisms can play a positive role on

characteristics of the product, as is the case of sour beers, where some specific metabolic traits will
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impart very particular complexity in flavor, aroma and acidity (Spitaels et al., 2014; Tonsmeire,
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2014; Verachtert & Derdelinckx, 2014).

Previously, bottle conditioning was the only choice for carbonating beers until the advent of

impermeable and pressure-resistant fermentation/maturation tanks, allowing a secondary

fermentation and maturation as well (Goldammer, 2008). Today, bottle conditioning has several

sensory advantages, with the possibility of enhancing the flavor for the production of new

metabolites, including esters and alcohols, in addition to eliminating dissolved oxygen that could

contribute to precipitated deterioration of beer (Vanderhaegen et al., 2003). This is the case of

Gueuze beers, wherein young Lambics (up to one year maturation) confer complex sugars to the

mixture, and old Lambics (two years or more) “inoculate” new microorganisms – especially
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Dekkera/Brettanomyces – that can consume these sugars during maturation in the bottle, afford

natural carbonation, and modify the beer sensory profile (Verachtert & Derdelinckx, 2014). In any

case, it is possible to apply this technique to most beers by adding sugars to the bottles and,

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optionally, pure cultures of different yeast strains to increase the complexity of final product

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(Vanderhaegen et al., 2003).

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3.2. Bioflavoring

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The appeal of consumers for beers with enhanced and differentiated sensory profiles grows along

with the search for products without chemical additives. In this context, one of the concepts of

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bioflavoring comes up for the synthesis and transformation of flavor compounds by biological
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methods, including the proposal of non-conventional yeasts (Daenen, Saison, et al., 2008; Daenen,

Sterckx, et al., 2008; Vanderhaegen et al., 2003). Addition of new compounds and flavor by

biological methods meets modern consumer expectations, either by: (a) using pure culture of a
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specific yeast species; (b) sequential or co-inoculation with different microorganisms; (c) use of
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genetically modified organisms (GMOs); or also by (d) adding isolated enzymes produced by other

microorganisms (Vanderhaegen et al., 2003). In the context of this work, only the first two cases
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were considered and reviewed.


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The sequential inoculation of Pichia kluyveri in beer wort is an example, where the combined effect

of different hop varieties showed to improve often desired esters such as isoamyl acetate and

isobutyl acetate, generally only slightly increasing the ethyl acetate content, that displays an

unpleasant “solvent” character at higher concentrations (Sarens & Swiegers, 2014), so proving to be

a good methodology to bring new contributions to the aromatic improvement of beers. Yeasts of the

genus Williopsis (= Cyberlindnera), otherwise, seems to produce high concentrations of acetate

esters such as isoamyl acetate, and could be used to exploit this feature in the production of beer in

which fruity notes are a desirable sensory trait (P.-R. Lee, Ong, Yu, Curran, & Liu, 2010; Li, Yu,

Curran, & Liu, 2012).


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Intense glucosidase activity is another interesting aspect in the search for bioflavoring. Some

Dekkera/Brettanomyces strains present those enzymes, but W. anomalus has also an innate ability to

markedly produce extracellular β-glucosidase in media containing cellobiose. It was shown that such

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enzymatic activity is enhanced in presence of ethanol, and inhibited by glucose, fructose and sucrose,

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a suggestive trait for a final-stage inoculation as well as for bottle conditioning (Swangkeaw,

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Vichitphan, Butzke, & Vichitphan, 2009).

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One such promising example in this context is the use of starter co-cultures including T. delbrueckii.

In these conditions, it is possible to verify increased use of the yeast assimilable nitrogen (YAN) in

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the medium, correlated to more expressive levels of some compounds – namely phenyl ethyl acetate,
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ethyl hexanoate and ethyl octanoate – that can positively stamp complex fruity, floral and aniseed

character to beers (Canonico et al., 2016).


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3.3. Low calorie beers

Beers with a low calorie content have achieved greater interest due to obesity problems, especially in
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Western populations, which have accounted for a growing market segment (Yeo & Liu, 2014). Low

calorie beers can be brewed addressing some alternative production methods, particularly seeking to
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reduce the concentration of carbohydrates in the final product. This type of beer may be done by both
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(a) special mashing and collection of wort with great amount of fermentable sugars, or (b)

fermentative process of an ordinary wort under specific conditions (Matthews, Byrne, & Hennigan,

2001).

A feasible method is the "dilution" of wort carbohydrates, which unfortunately leads to poorly

structured beers, without much flavor expressiveness (Sato, Mizuno, Mukai, & Amano, 2006).

Another possible approach is the higher conversion of wort sugars during mashing, aiming higher

attenuation level in fermentation, thus lowering the sugar content in the final product (Matthews et

al., 2001). However, with more fermentable sugars in the wort, more alcohol will be released to the

medium. Since ethanol has a higher energy value in comparison to carbohydrates, the production of
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low calorie beers will not be possible unless part of the alcohol is later removed (Yeo & Liu, 2014).

It is possible to drive fermentations by inoculating microorganisms with ability to hydrolyze and

assimilate more complex sugars, thus reducing the concentration of residual sugars in the beer – as is

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the case of Brettanomyces/Dekkera, that lead to low-calorie, slightly more alcoholic beers (Steensels

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et al., 2015). Another possibility would be to increase the concentration of those carbohydrates

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which are not assimilated nor by yeasts, or by the human body, as is the case of β-glucans and

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arabinoxylans (Yeo & Liu, 2014).

3.4. Low alcohol beers

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An approach that is becoming more popular in brewing is the production of beers with reduced
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alcohol content, considering two aspects that deserve attention: health and physical integrity of

consumers. Methods to obtain beers with reduced alcohol content may be both physical and

biological. In the first case, it may be done by thermal techniques (rectification; evaporation) or
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membrane usage (dialysis; reverse osmosis). Methods employing biological factors may rely on
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traditional brewery plant (limited fermentation; changed mashing; special yeasts), or in the use of

specific machinery (continuous fermentation) (Brányik, Silva, Baszczyňski, Lehnert, & Almeida e
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Silva, 2012).
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Hence, new methods have been studied with the aim of producing beverages with lower alcohol

content, but keeping a superior sensory quality. In this sense, non-Saccharomyces yeasts represent a

very attractive alternative to produce low alcohol beers, since this technique does not entail

processing extra costs. In addition, it has the advantage of avoiding the involuntary extraction of

flavor compounds, a common problem when ethanol extraction is carried out after fermentation by

distillation, reverse osmosis or dialysis (Erten & Campbell, 2001).

Wickerhamomyces subpelliculosus (= Pichia subpelliculosa) and Cyberlindnera saturnus (=

Williopsis saturnus), for example, showed interesting results in the production of low alcohol wines

with acceptable flavor profiles (Erten & Campbell, 2001),. Those latter abilities are also related to
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alcohol acetyltransferase and esterase enzymes that enhance production of aromatic isoamyl acetate

and ethyl acetate (Erten & Tanguler, 2010; P.-R. Lee et al., 2010; Plata, Millán, Mauricio, & Ortega,

2003). The use of those yeasts in winemaking was possible by agitation and aeration of the must

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during alcoholic fermentation suggesting its application in brewing, even in mixed or sequential

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inoculation with Saccharomyces.

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Many strains of T. delbrueckii are only able to metabolize glucose, fructose, and sucrose, so that they

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cannot degrade maltose, maltotriose, and other complex carbohydrates. It should be an interesting

way, once the resulting beer would be much lower in alcohol content, usually close to 0.9 % (v/v),

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with the advantage that many of those strains impart rich fruity flavor and aroma to the beer (Michel
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et al., 2016). Undoubtedly, the use of T. delbrueckii represents a hopeful approach in this context,

once pure cultures of this species seem to reduce up to 50 % the alcoholic content, and

simultaneously contribute to a complex fruity character in the final beer (Canonico et al., 2016).
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In this context, it is also possible to consider the use of cold contact process in restrained
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fermentations, a methodology that allies very low temperatures (~ 0 ºC) to long fermentation period.

Selected cold-resistant strains are obviously necessary, and basal metabolism lead to ester, higher
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alcohol, and carbonyl reduction, but virtually very little ethanol production in low-density worts.
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Despite obtaining low/free-alcohol beers, this method enables to obtain a high volatile profile, and

low aldehyde reduction (Brányik et al., 2012; Perpète & Collin, 1999, 2000).

3.5. Functional beers

Functional beers – with health benefits for those who consume them moderately – point to the use of

non-conventional yeasts due to the potential these microorganisms may play in production or

transformation of some beneficial compounds. The concept of functional beer is necessarily

combined to a low alcohol product, and to a matrix with substantial amounts of fibers, vitamins,

minerals and polyphenols (Yeo & Liu, 2014).


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This is the case of melatonin is the sleep regulating hormone in mammals and is a molecule that

shows antioxidant properties. It is produced in ever-smaller amounts with human increasing age, and

in such case, it can be produced in beer during alcoholic fermentation by appropriate yeasts,

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constituting an exogenous source of this hormone, so aggregating antioxidant, anti-aging, anti-

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inflammatory, antitumor and immunomodulatory properties to the beer (Maldonado, Moreno, &

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Calvo, 2009; Rodriguez-Naranjo, Gil-Izquierdo, Troncoso, Cantos-Villar, & Garcia-Parrilla, 2011).

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Besides having the aforesaid advantages, beers that undergo natural carbonation also present better

nutritional properties (García-Moreno, Calvo, & Maldonado, 2013). To date, however, no specific

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study attempted to relate fermentations driven by non-Saccharomyces yeasts with the production of
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functional beers, pointing the opportunity for future researches.

4. Conclusions

Many yeast species are emerging as candidates for the production of specialty beers. Innovation not
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only relies on obtaining new products with more complex aromatic and flavor character, but also on
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increasing the range of approaches to achieve different results in a growing market. Using non-

Saccharomyces yeasts for pitching, in sequential or co-inoculation, in bottle conditioning, or many


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other strategies, has presented the possibility to unveil old and new methods to produce specialty and
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original beers. Brewers are now reinterpreting the role of different yeast species – regarded as

spoilage microorganisms in uncontrolled or without good manufacturing practices – and learning

with them to find a multitude of applications and benefits in addition to raw material. Yeasts from

the genera Dekkera, Hanseniaspora, Pichia, Torulaspora, Wickerhamomyces, and others, can offer a

diversified enzymatic apparatus and bioconversion abilities that are allowing brewers to work with

new concepts that include bioflavoring, likewise beers with reduced calorie and alcohol contents, or

even functional beers. Nonetheless, the introduction of a new yeast must be previously calculated,

because each microorganism is unique, and can develop a range of adaptations in contact with

different substrates or conditions. Finally, it is important not to ignore the importance of recent craft
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brewery revolutions occurring in many parts of the globe, a movement that has been paving the way

to explore such traits and possibilities in order to find a growing market of enthusiast and expert

consumers.

P T
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Table 1 Number and growth of microbreweries in Europe (2009 – 2014)

microbreweries relative
2009 2014 growth (%)
Spain 27 314 1062
Norway 13 65 400

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Sweden 30 149 396

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Czech Republic 51 238 366

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Slovakia 9 39 333
Slovenia 20 49 145
Italy 242 585 141

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France 263 566 115
United Kingdom 694 1414 103
Switzerland 232 440 89

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Adapted from: Beers Statistics 2015 (The Brewers of Europe)
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Table 2 Taxonomical status, isolation sources and use of non-conventional yeasts for brewing

T
IP
species anamorph a synonyms a isolation sources b application food safety c

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Cyberlindnera saturnus Williopsis saturnus; insect, soil bioflavoring; low
Saccharomyces saturnus alcohol beer safe

S
Dekkera anomala Brettanomyces anomalus Dekkera/Brettanomyces beer, cider, soft drink bioflavoring; low

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safe
claussenii calorie beer
Dekkera bruxellensis Brettanomyces bruxellensis Brettanomyces lambicus; beer, grape, wine, bioflavoring; low
safe

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Brettanomyces custersii ginger ale calorie beer
Hanseniaspora uvarum Kloeckera apiculata Kloeckeraspora uvarum; grape must, fruits, soil, bioflavoring
Hanseniaspora apiculata sour dough, insect, safe

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flowers
Pichia kluyveri Hansenula kluyveri olives, fruits, flowers, bioflavoring
safe
soil
Torulaspora delbrueckii Candida colliculosa
PT Saccharomyces rosei;
Saccharomyces delbrueckii
beer, fruits, soil, milk,
moss, mushroom
bioflavoring; low
alcohol beer
safe
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Wickerhamomyces subpelliculosus Pichia subpelliculosa; fruits; vegetables; low alcohol beer
safe
Hansenula subpelliculosa molasses; jam; milk
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Wickerhamomyces anomalus Candida beverwijkiae Pichia anomala; beer, sour dough, fruits, bioflavoring; low
Saccharomyces anomalus; sake, wine, oak calorie beer safe
Hansenula anomala
a
Taxonomical records according to National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/guide/taxonomy), and Index Fungorum
(http://www.indexfungorum.org). b Data from Global Catalogue of Microorganisms (http://gcm.wfcc.info). c According to Bourdichon et al. (2012)
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Table 3 Major aromatic compounds, enzymes and peculiar traits of non-conventional yeasts in brewing

T
aromatic prominent
species flavor compound disadvantage peculiar traits references
descriptor enzyme

IP
C. saturnus isoamyl acetate banana, pear AAT biofilm formation ethanol sensitive (Erten & Campbell, 2001; Erten &

CR
ethyl acetate fruity, solvent Tanguler, 2010; P.-R. Lee et al.,
2010)
D. anomala acetic acid vinegary BGL, CDC, high acetic acid osmotolerant; pH tolerant; (Lentz & Harris, 2015)

S
ehtyl phenols clove; leather VPR production; overall Custer effect
ethyl esters fruity off-flavor

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D. bruxellensis acetic acid vinegary BGL, CDC, high acetic acid osmotolerant; pH tolerant; (Lentz & Harris, 2015)
ehtyl phenols clove; leather VPR production; overall Custer effect
ethyl esters fruity off-flavor

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H. uvarum isoamyl acetate banana; pear BGL cycloheximide-resistant; (Arévalo Villena, Úbeda Iranzo,
ethyl acetate fruity; solvent ethanol tolerant Cordero Otero, & Briones Pérez,
2005; Spitaels et al., 2014; Wang,
Mas, & Esteve-Zarzoso, 2015)

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P. kluyveri isoamyl acetate banana; pear AAT osmotolerant; Pasteur (Fredlund, 2004; Ma, He, Cao, Bai,
isobutyl acetate banana; fruity effect; pH tolerant & Li, 2016; Sarens & Swiegers,
2014; Steensels & Verstrepen, 2014;

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Walker, 2011)
T. delbrueckii isoamyl acetate banana; pear AAT, EST, high acetaldehyde osmotolerant; (Canonico et al., 2016; Charoenchai,
ethyl butyrate banana; apple; INV, BGL production cryotolerant; ethanol Fleet, Henschke, & Todd, 1997;
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strawberry tolerant Peinado, Moreno, Bueno, Moreno,
acetaldehyde green apple & Mauricio, 2004; Plata et al., 2003)
W. subpelliculosus isoamyl acetate banana, pear AAT, EST biofilm formation; (Erten & Campbell, 2001; Plata et
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ethyl acetate fruity; solvent excessive ester al., 2003)


production
W. anomalus ethyl acetate fruity; solvent EST, BGL biofilm formation ethanol sensitive; (Charoenchai et al., 1997; Erten &
osmotolerant; pH tolerant Campbell, 2001; Passoth et al.,
2006)
Alcohol acetyltransferase (AAT), Esterase (EST), Invertase (INV), β-glucosidase (BGL), Cinnamate decarboxylase (CDC), Vinylphenol
reductase (VPR).
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References

Aguilar-Uscanga, M. G., Délia, M. L., & Strehaiano, P. (2003). Brettanomyces bruxellensis: effect of

oxygen on growth and acetic acid production. Applied Microbiology and Biotechnology,

T
61(2), 157-162. doi: 10.1007/s00253-002-1197-z

P
Albertin, W., Chasseriaud, L., Comte, G., Panfili, A., Delcamp, A., Salin, F., . . . Bely, M. (2014).

RI
Winemaking and bioprocesses strongly shaped the genetic diversity of the ubiquitous yeast

SC
Torulaspora delbrueckii. PLoS ONE, 9(4), e94246. doi: 10.1371/journal.pone.0094246

Alves-Araújo, C., Pacheco, A., Almeida, M. J., Spencer-Martins, I., Leão, C., & Sousa, M. J. (2007).

NU
Sugar utilization patterns and respiro-fermentative metabolism in the baker's yeast
MA
Torulaspora delbrueckii. Microbiology, 153(3), 898-904. doi:

doi:10.1099/mic.0.2006/003475-0

Arévalo Villena, M., Úbeda Iranzo, J. F., Cordero Otero, R. R., & Briones Pérez, A. I. (2005).
ED

Optimization of a rapid method for studying the cellular location of β-glucosidase activity in
PT

wine yeasts. Journal of Applied Microbiology, 99(3), 558-564. doi: 10.1111/j.1365-

2672.2005.02627.x
CE

Barata, A., Caldeira, J., Botelheiro, R., Pagliara, A., Malfeito-Ferreira, M., & Loureiro, V. (2008).
AC

Survival patterns of Dekkera bruxellensis in wines and inhibitory effect of sulphur dioxide.

[10.1016/j.ijfoodmicro.2007.11.020]. International Journal of Food Microbiology, 121, 201-

207. doi: 10.1016/j.ijfoodmicro.2007.11.020

Barnett, J. A., & Entian, K. D. (2005). A history of research on yeasts. 9: Regulation of sugar

metabolism. Yeast, 22(11), 835-894.

Bely, M., Stoeckle, P., Masneuf-Pomarède, I., & Dubourdieu, D. (2008). Impact of mixed

Torulaspora delbrueckii–Saccharomyces cerevisiae culture on high-sugar fermentation.

International Journal of Food Microbiology, 122(3), 312-320. doi:

http://dx.doi.org/10.1016/j.ijfoodmicro.2007.12.023
ACCEPTED MANUSCRIPT

Bokulich, N. A., & Bamforth, C. W. (2013). The microbiology of malting and brewing.

Microbiology and Molecular Biology Reviews, 77(2), 157-172. doi: 10.1128/mmbr.00060-12

Bokulich, N. A., Bamforth, C. W., & Mills, D. A. (2012). Brewhouse-resident microbiota are

T
responsible for multi-stage fermentation of American Coolship Ale. PLoS ONE, 7(4),

P
e35507. doi: 10.1371/journal.pone.0035507

RI
Bourdichon, F., Casaregola, S., Farrokh, C., Frisvad, J. C., Gerds, M. L., Hammes, W. P., . . .

SC
Hansen, E. B. (2012). Food fermentations: Microorganisms with technological beneficial use.

International Journal of Food Microbiology, 154(3), 87-97. doi:

NU
http://dx.doi.org/10.1016/j.ijfoodmicro.2011.12.030
MA
Brandam, C., Castro-Martínez, C., Délia, M.-L., Ramón-Portugal, F., & Strehaiano, P. (2007). Effect

of temperature on Brettanomyces bruxellensis: metabolic and kinetic aspects. Canadian

Journal of Microbiology, 54(1), 11-18. doi: 10.1139/W07-126


ED

Brányik, T., Silva, D. P., Baszczyňski, M., Lehnert, R., & Almeida e Silva, J. B. (2012). A review of
PT

methods of low alcohol and alcohol-free beer production. Journal of Food Engineering,

108(4), 493-506. doi: http://dx.doi.org/10.1016/j.jfoodeng.2011.09.020


CE

Brewers-Association. (2016). National beer sales and production data. The New Brewer Retrieved
AC

March, 21, 2016, from https://www.brewersassociation.org/statistics/national-beer-sales-

production-data/

Canonico, L., Agarbati, A., Comitini, F., & Ciani, M. (2016). Torulaspora delbrueckii in the brewing

process: A new approach to enhance bioflavour and to reduce ethanol content. Food

Microbiology, 56, 45-51. doi: 10.1016/j.fm.2015.12.005

Charoenchai, C., Fleet, G. H., Henschke, P. A., & Todd, B. E. N. T. (1997). Screening of non-

Saccharomyces wine yeasts for the presence of extracellular hydrolytic enzymes. Australian

Journal of Grape and Wine Research, 3(1), 2-8. doi: 10.1111/j.1755-0238.1997.tb00109.x


ACCEPTED MANUSCRIPT

Chatonnet, P., Dubourdie, D., Boidron, J. N. l., & Pons, M. (1992). The origin of ethylphenols in

wines. [10.1002/jsfa.2740600205]. Journal of the Science of Food and Agriculture, 60(2),

165-178.

T
Ciani, M., & Comitini, F. (2011). Non-Saccharomyces wine yeasts have a promising role in

P
biotechnological approaches to winemaking. Annals of Microbiology, 61(1), 25-32.

RI
Conterno, L., Aprea, E., Franceschi, P., Viola, R., & Vrhovsek, U. (2013). Overview of Dekkera

SC
bruxellensis behaviour in an ethanol-rich environment using untargeted and targeted

metabolomic approaches. Food Research International, 51(2), 670-678. doi:

NU
10.1016/j.foodres.2013.01.049
MA
Cordero-Bueso, G., Esteve-Zarzoso, B., Cabellos, J., Gil-Díaz, M., & Arroyo, T. (2013).

Biotechnological potential of non-Saccharomyces yeasts isolated during spontaneous

fermentations of Malvar (Vitis vinifera cv. L.). European Food Research and Technology,
ED

236(1), 193-207. doi: 10.1007/s00217-012-1874-9


PT

Cortés-Diéguez, S., Rodriguez-Solana, R., Domínguez, J. M., & Díaz, E. (2015). Impact odorants

and sensory profile of young red wines from four Galician (NW of Spain) traditional
CE

cultivars. Journal of the Institute of Brewing, 121(4), 628-635. doi: 10.1002/jib.252


AC

Crauwels, S., Steensels, J., Aerts, G., Willems, K. A., Verstrepen, K. J., & Lievens, B. (2015).

Brettanomyces bruxellensis, essential contributor in spontaneous beer fermentations

providing novel opportunities for the brewing industry. Brewing Science,

68(September/October), 110-121.

Daenen, L., Saison, D., Sterckx, F., Delvaux, F. R., Verachtert, H., & Derdelinckx, G. (2008).

Screening and evaluation of the glucoside hydrolase activity in Saccharomyces and

Brettanomyces brewing yeasts. Journal of Applied Microbiology, 104(2), 478-488. doi:

10.1111/j.1365-2672.2007.03566.x
ACCEPTED MANUSCRIPT

Daenen, L., Sterckx, F., Delvaux, F. R., Verachtert, H., & Derdelinckx, G. (2008). Evaluation of the

glycoside hydrolase activity of a Brettanomyces strain on glycosides from sour cherry

(Prunus cerasus L.) used in the production of special fruit beers. FEMS Yeast Research, 8(7),

T
1103-1114. doi: 10.1111/j.1567-1364.2008.00421.x

P
De Keersmaecker, J. (1996). The mystery of lambic beer. Scientific American 275, 74-81.

RI
Di Maro, E., Ercolini, D., & Coppola, S. (2007). Yeast dynamics during spontaneous wine

SC
fermentation of the Catalanesca grape. International Journal of Food Microbiology, 117(2),

201-210. doi: 10.1016/j.ijfoodmicro.2007.04.007

NU
Erten, H., & Campbell, I. (2001). The production of low-alcohol wines by aerobic yeasts. Journal of
MA
the Institute of Brewing, 59(3), 207-215. doi: 10.1002/j.2050-0416.1953.tb06929.x

Erten, H., & Tanguler, H. (2010). Influence of Williopsis saturnus yeasts in combination with

Saccharomyces cerevisiae on wine fermentation. Letters in Applied Microbiology, 50(5),


ED

474-479. doi: 10.1111/j.1472-765X.2010.02822.x


PT

Etschmann, M., Huth, I., Walisko, R., Schuster, J., Krull, R., Holtmann, D., . . . Schrader, J. (2015).

Improving 2‐phenylethanol and 6‐pentyl‐α‐pyrone production with fungi by


CE

microparticle‐enhanced cultivation (MPEC). Yeast, 32(1), 145-157. doi: 10.1002/yea.3022


AC

Fredlund, E. (2004). Central carbon metabolism of the biocontrol yeast Pichia anomala: Influence of

oxygen limitation. PhD Doctoral thesis, Swedish University of Agricultural Sciences,

Uppsala. (488)

Fredlund, E., Druvefors, U. Ä., Olstorpe, M. N., Passoth, V., & Schnürer, J. (2004). Influence of

ethyl acetate production and ploidy on the anti-mould activity of Pichia anomala. FEMS

Microbiology Letters, 238(1), 133-137. doi: 10.1111/j.1574-6968.2004.tb09747.x

García-Moreno, H., Calvo, J. R., & Maldonado, M. D. (2013). High levels of melatonin generated

during the brewing process. Journal of Pineal Research, 55(1), 26-30. doi: 10.1111/jpi.12005
ACCEPTED MANUSCRIPT

Goldammer, T. (2008). The brewers' handbook. The complete book to brewing beer (2 ed.). Clifton:

Apex Publishers.

González, R., Quirós, M., & Morales, P. (2013). Yeast respiration of sugars by non-Saccharomyces

T
yeast species: A promising and barely explored approach to lowering alcohol content

P
of wines. Trends in Food Science & Technology, 29(1), 55-61. doi:

RI
10.1016/j.tifs.2012.06.015

SC
Hanl, L., Sommer, P., & Arneborg, N. (2004). The effect of decreasing oxygen feed rates on growth

and metabolism of Torulaspora delbrueckii. [journal article]. Applied Microbiology and

NU
Biotechnology, 67(1), 113-118. doi: 10.1007/s00253-004-1695-2
MA
Harris, V., Ford, C., Jiranek, V., & Grbin, P. (2009). Survey of enzyme activity responsible for

phenolic off-flavour production by Dekkera and Brettanomyces yeast. Applied Microbiology

and Biotechnology, 81(6), 1117-1127.


ED

Hayashi, N., Arai, R., Tada, S., Taguchi, H., & Ogawa, Y. (2007). Detection and identification of
PT

Brettanomyces/Dekkera sp. yeasts with a loop-mediated isothermal amplification method.

Food Microbiology, 24, 778-785.


CE

Hernández-Orte, P., Cersosimo, M., Loscos, N., Cacho, J., Garcia-Moruno, E., & Ferreira, V. (2008).
AC

The development of varietal aroma from non-floral grapes by yeasts of different genera. Food

Chemistry, 107(3), 1064-1077. doi: 10.1016/j.foodchem.2007.09.032

Hornsey, I. (2007). The Chemistry and Biology of winemaking. Cambridge: The Royal Society of

Chemistry.

Johnson, E. (2013). Biotechnology of non-Saccharomyces yeasts - the ascomycetes. Applied

Microbiology and Biotechnology, 97(2), 503-517. doi: 10.1007/s00253-012-4497-y

Kell, J. (2016). What You Didn't Know About the Boom in Craft Beer Retrieved 24/05/2016, 2016,

from http://fortune.com/2016/03/22/craft-beer-sales-rise-2015/
ACCEPTED MANUSCRIPT

Kumara, H. M. C. S., De Cort, S., & Verachtert, H. (1993). Localization and characterization of α-

glucosidase activity in Brettanomyces lambicus. Applied and Environmental Microbiology,

59(8), 2352-2358.

T
Kumara, H. M. C. S., & Verachtert, H. (1991). Identification of lambic supperattenuating micro-

P
organisms by the use of selective antibiotics. Journal of the Institute of Brewing, 97(3), 181-

RI
185. doi: 10.1002/j.2050-0416.1991.tb01064.x

SC
Kurtzman, C. (2011). Phylogeny of the ascomycetous yeasts and the renaming of Pichia anomala to

Wickerhamomyces anomalus Antonie van Leeuwenhoek, 99(1), 13-23.

NU
Lee, P.-R., Ong, Y.-L., Yu, B., Curran, P., & Liu, S.-Q. (2010). Evolution of volatile compounds in
MA
papaya wine fermented with three Williopsis saturnus yeasts. International Journal of Food

Science & Technology, 45(10), 2032-2041. doi: 10.1111/j.1365-2621.2010.02369.x

Lee, Y.-J., Choi, Y.-R., Lee, S.-Y., Park, J.-T., Shim, J.-H., Park, K.-H., & Kim, J.-W. (2011).
ED

Screening wild yeast strains for alcohol fermentation from various fruits. Mycobiology, 39(1),
PT

33-39. doi: 10.4489/MYCO.2011.39.1.033

Lentz, M., & Harris, C. (2015). Analysis of growth inhibition and metabolism of hydroxycinnamic
CE

acids by brewing and spoilage strains of Brettanomyces yeast. Foods, 4(4), 581-593.
AC

Li, X., Yu, B., Curran, P., & Liu, S.-Q. (2012). Impact of two Williopsis yeast strains on the volatile

composition of mango wine. International Journal of Food Science & Technology, 47(4),

808-815. doi: 10.1111/j.1365-2621.2011.02912.x

Lodolo, E. J., Kock, J. L. F., Axcell, B. C., & Brooks, M. (2008). The yeast Saccharomyces

cerevisiae – the main character in beer brewing. FEMS Yeast Research, 8(7), 1018-1036. doi:

10.1111/j.1567-1364.2008.00433.x

Ma, C., He, Y., Cao, Y., Bai, X., & Li, H. (2016). Analysis of flavour compounds in beer with

extruded sorghum as an adjunct using headspace solid-phase micro-extraction and gas


ACCEPTED MANUSCRIPT

chromatography–mass spectrometry. Journal of the Institute of Brewing, 122(2), 251-260.

doi: 10.1002/jib.330

Maldonado, M. D., Moreno, H., & Calvo, J. R. (2009). Melatonin present in beer contributes to

T
increase the levels of melatonin and antioxidant capacity of the human serum. Clinical

P
Nutrition, 28(2), 188-191. doi: http://dx.doi.org/10.1016/j.clnu.2009.02.001

RI
Masneuf-Pomarede, I., Bely, M., Marullo, P., & Albertin, W. (2015). The genetics of non-

SC
conventional wine yeasts: current knowledge and future challenges. Frontiers in

Microbiology, 6. doi: 10.3389/fmicb.2015.01563

NU
Matthews, S. L., Byrne, H., & Hennigan, G. P. (2001). Preparation of a low carbohydrate beer by
MA
mashing at high temperature with glucoamylase. Journal of the Institute of Brewing, 107(3),

185-194. doi: 10.1002/j.2050-0416.2001.tb00090.x

Meilgaard, M. C. (1982). Prediction of flavor differences between beers from their chemical
ED

composition. Journal of Agricultural and Food Chemistry, 30(6), 1009-1017. doi:


PT

10.1021/jf00114a002

Michel, M., Kopecká, J., Meier-Dörnberg, T., Zarnkow, M., Jacob, F., & Hutzler, M. (2016).
CE

Screening for new brewing yeasts in the non-Saccharomyces sector with Torulaspora
AC

delbrueckii as model. Yeast, 33(4), 129-144. doi: 10.1002/yea.3146

Papazian, C. (2003). The complete joy of homebrewing (3 ed.). New York: Harper Collins.

Passoth, V., Fredlund, E., Druvefors, U. Ä., & Schnürer, J. (2006). Biotechnology, physiology and

genetics of the yeast Pichia anomala. FEMS Yeast Research, 6(1), 3-13. doi: 10.1111/j.1567-

1364.2005.00004.x

Peinado, R. A., Moreno, J., Bueno, J. E., Moreno, J. A., & Mauricio, J. C. (2004). Comparative study

of aromatic compounds in two young white wines subjected to pre-fermentative

cryomaceration. Food Chemistry, 84(4), 585-590. doi: 10.1016/S0308-8146(03)00282-6


ACCEPTED MANUSCRIPT

Perpète, P., & Collin, S. (1999). Fate of the worty flavours in a cold contact fermentation. Food

Chemistry, 66(3), 359-363. doi: http://dx.doi.org/10.1016/S0308-8146(99)00085-0

Perpète, P., & Collin, S. (2000). How to improve the enzymatic worty flavour reduction in a cold

T
contact fermentation. Food Chemistry, 70(4), 457-462. doi: http://dx.doi.org/10.1016/S0308-

P
8146(00)00111-4

RI
Pires, E., Teixeira, J., Brányik, T., & Vicente, A. (2014). Yeast: the soul of beer’s aroma - a review

SC
of flavour-active esters and higher alcohols produced by the brewing yeast. Applied

Microbiology and Biotechnology, 98(5), 1937-1949. doi: 10.1007/s00253-013-5470-0

NU
Piškur, J., Ling, Z., Marcet-Houben, M., Ishchuk, O. P., Aerts, A., LaButti, K., . . . Phister, T. (2012).
MA
The genome of wine yeast Dekkera bruxellensis provides a tool to explore its food-related

properties. International Journal of Food Microbiology, 157(2), 202-209. doi:

http://dx.doi.org/10.1016/j.ijfoodmicro.2012.05.008
ED

Plata, C., Millán, C., Mauricio, J. C., & Ortega, J. M. (2003). Formation of ethyl acetate and isoamyl
PT

acetate by various species of wine yeasts. Food Microbiology, 20(2), 217-224. doi:

http://dx.doi.org/10.1016/S0740-0020(02)00101-6
CE

Portugal, C., Sáenz, Y., Rojo-Bezares, B., Zarazaga, M., Torres, C., Cacho, J., & Ruiz-Larrea, F.
AC

(2014). Brettanomyces susceptibility to antimicrobial agents used in winemaking: in vitro and

practical approaches. European Food Research and Technology, 238(4), 641-652. doi:

10.1007/s00217-013-2143-2

Renouf, V., & Lonvaud-Funel, A. (2007). Development of an enrichment medium to detect

Dekkera/Brettanomyces bruxellensis, a spoilage wine yeast, on the surface of grape berries.

Microbiological Research, 162(2), 154-167.

Rodriguez-Naranjo, M. I., Gil-Izquierdo, A., Troncoso, A. M., Cantos-Villar, E., & Garcia-Parrilla,

M. C. (2011). Melatonin is synthesised by yeast during alcoholic fermentation in wines. Food

Chemistry, 126(4), 1608-1613. doi: http://dx.doi.org/10.1016/j.foodchem.2010.12.038


ACCEPTED MANUSCRIPT

Rogers, C. M., Veatch, D., Covey, A., Staton, C., & Bochman, M. L. (2016). Terminal acidic shock

inhibits sour beer bottle conditioning by Saccharomyces cerevisiae. Food Microbiology, 57,

151-158. doi: http://dx.doi.org/10.1016/j.fm.2016.02.012

T
Rojas, V., Gil, J. V., Manzanares, P., Gavara, R., Piñaga, F., & Flors, A. (2002). Measurement of

P
alcohol acetyltransferase and ester hydrolase activities in yeast extracts. Enzyme and

RI
Microbial Technology, 30(2), 224-230. doi: 10.1016/S0141-0229(01)00483-5

SC
Rozpędowska, E., Hellborg, L., Ishchuk, O. P., Orhan, F., Galafassi, S., Merico, A., . . . Piskur, J.

(2011). Parallel evolution of the make-accumulate-consume strategy in Saccharomyces and

NU
Dekkera yeasts. Nature Communications, 2(302), 1-7. doi: 10.1038/ncomms1305
MA
Salvadó, Z., Arroyo-López, F. N., Guillamón, J. M., Salazar, G., Querol, A., & Barrio, E. (2011).

Temperature adaptation markedly determines evolution within the genus Saccharomyces.

Applied and Environmental Microbiology, 77(7), 2292-2302. doi: 10.1128/aem.01861-10


ED

Sarens, S., & Swiegers, J. H. (2014). United States Patent No. US20140234480A1.
PT

Sato, K., Mizuno, A., Mukai, N., & Amano, H. (2006). United States Patent No.: US7115289.

Schneider, J., Rupp, O., Trost, E., Jaenicke, S., Passoth, V., Goesmann, A., . . . Brinkrolf, K. (2012).
CE

Genome sequence of Wickerhamomyces anomalus DSM 6766 reveals genetic basis of


AC

biotechnologically important antimicrobial activities. FEMS Yeast Research, 12(3), 382-386.

doi: 10.1111/j.1567-1364.2012.00791.x

Sicard, D., & Legras, J.-L. (2011). Bread, beer and wine: Yeast domestication in the Saccharomyces

sensu stricto complex. Comptes Rendus Biologies, 334(3), 229-236. doi:

10.1016/j.crvi.2010.12.016

Spaepen, M., Van Oevelen, D., & Verachtert, H. (1978). Fatty acids and esters produced during the

spontaneous fermentation of Lambic and Gueuze. Journal of the Institute of Brewing, 84(5),

278-282. doi: 10.1002/j.2050-0416.1978.tb03888.x


ACCEPTED MANUSCRIPT

Spaepen, M., & Verachtert, H. (1982). Esterase activity in the genus Brettanomyces. Journal of the

Institute of Brewing, 88(1), 11-17. doi: 10.1002/j.2050-0416.1982.tb04061.x

Spitaels, F., Wieme, A. D., Janssens, M., Aerts, M., Daniel, H.-M., Van Landschoot, A., . . .

T
Vandamme, P. (2014). The microbial diversity of traditional spontaneously fermented lambic

P
beer. PLoS ONE, 9(4), 1-13. doi: 10.1371/journal.pone.0095384

RI
Steensels, J., Daenen, L., Malcorps, P., Derdelinckx, G., Verachtert, H., & Verstrepen, K. J. (2015).

SC
Brettanomyces yeasts - From spoilage organisms to valuable contributors to industrial

fermentations. International Journal of Food Microbiology, 206, 24-38. doi:

NU
10.1016/j.ijfoodmicro.2015.04.005
MA
Steensels, J., & Verstrepen, K. J. (2014). Taming wild yeast: potential of conventional and

nonconventional yeasts in industrial fermentations. Annual Review of Microbiology, 68(1),

61-80. doi: 10.1146/annurev-micro-091213-113025


ED

Swangkeaw, J., Vichitphan, S., Butzke, C., & Vichitphan, K. (2009). The characterisation of a novel
PT

Pichia anomala β-glucosidase with potentially aroma-enhancing capabilities in wine. Annals

of Microbiology, 59(2), 335-343. doi: 10.1007/BF03178336


CE

Tataridis, P., Kanelis, A., Logotetis, S., & Nerancis, E. (2013). Use of non-Saccharomyces
AC

Torulaspora delbrueckii yeast strains in winemaking and brewing. Journal of Natural

Science(124), 415-426. doi: 10.2298/ZMSPN1324415T

Thomson, J. M., Gaucher, E. A., Burgan, M. F., De Kee, D. W., Li, T., Aris, J. P., & Benner, S. A.

(2005). Resurrecting ancestral alcohol dehydrogenases from yeast. Nature genetics, 37(6),

630-635.

Tonsmeire, M. (2014). American sour beer: innovative techniques for mixed fermentations. Boulder:

Brewers Publications.

van Dijken, H. (2002). Biochemistry, genetics, biotechnology and ecology of non-conventional yeasts

(NCY). Paper presented at the The 21st International Specialized Symposium on Yeasts Lviv
ACCEPTED MANUSCRIPT

National University. Conference report retrieved from

https://femsyr.oxfordjournals.org/content/1/4/337

Vanbeneden, N., Gils, F., Delvaux, F., & Delvaux, F. R. (2008). Formation of 4-vinyl and 4-ethyl

T
derivatives from hydroxycinnamic acids: Occurrence of volatile phenolic flavour compounds

P
in beer and distribution of Pad1-activity among brewing yeasts. Food Chemistry, 107(1),

RI
221-230. doi: 10.1016/j.foodchem.2007.08.008

SC
Vanbeneden, N., Van Roey, T., Willems, F., Delvaux, F., & Delvaux, F. R. (2008). Release of

phenolic flavour precursors during wort production: Influence of process parameters and grist

NU
composition on ferulic acid release during brewing. Food Chemistry, 111(1), 83-91. doi:

10.1016/j.foodchem.2008.03.029
MA
Vanderhaegen, B., Neven, H., Coghe, S., Verstrepen, K. J., Derdelinckx, G., & Verachtert, H.

(2003). Bioflavoring and beer refermentation. Applied Microbiology and Biotechnology,


ED

62(2-3), 140-150. doi: 10.1007/s00253-003-1340-5


PT

Verachtert, H., & Derdelinckx, G. (2014). Belgian acidic beers: daily reminiscences of the past.

Cerevisia, 38(4), 121-128. doi: 10.1016/j.cervis.2014.04.002


CE

Walker, G. (2011). Pichia anomala: cell physiology and biotechnology relative to other yeasts.
AC

Antonie van Leeuwenhoek, 99(1), 25-34. doi: 10.1007/s10482-010-9491-8

Wang, C., Mas, A., & Esteve-Zarzoso, B. (2015). Interaction between Hanseniaspora uvarum and

Saccharomyces cerevisiae during alcoholic fermentation. International Journal of Food

Microbiology, 206, 67-74. doi: 10.1016/j.ijfoodmicro.2015.04.022

White, C., & Zainasheff, J. (2010). Yeast: The practical guide to beer fermentation. Boulder:

Brewers Publications.

Winterhalter, P., & Skouroumounis, G. K. (1997). Glycoconjugated aroma compounds: Occurrence,

role and biotechnological transformation. In R. G. Berger, W. Babel, H. W. Blanch, C. L.

Cooney, S. O. Enfors, K. E. L. Eriksson, A. Fiechter, A. M. Klibanov, B. Mattiasson, S. B.


ACCEPTED MANUSCRIPT

Primrose, H. J. Rehm, P. L. Rogers, H. Sahm, K. Schügerl, G. T. Tsao, K. Venkat, J.

Villadsen, U. Stockar & C. Wandrey (Eds.), Biotechnology of Aroma Compounds (pp. 73-

105). Berlin, Heidelberg: Springer Berlin Heidelberg.

T
Woolfit, M., Rozpędowska, E., Piškur, J., & Wolfe, K. H. (2007). Genome survey sequencing of the

P
wine spoilage yeast Dekkera (Brettanomyces) bruxellensis. Eukaryotic Cell, 6(4), 721-733.

RI
doi: 10.1128/ec.00338-06

SC
Yeo, H. Q., & Liu, S.-Q. (2014). An overview of selected specialty beers: developments, challenges

and prospects. International Journal of Food Science & Technology, 49(7), 1607-1618. doi:

NU
10.1111/ijfs.12488
MA
Yoshioka, K., & Hashimoto, N. (1981). Ester formation by alcohol acetyltransferase from brewers’

yeast. Agricultural and Biological Chemistry, 45(10), 2183-2190.

Yu, J., Vasanthan, T., & Temelli, F. (2001). Analysis of phenolic acids in barley by high-
ED

performance liquid chromatography. Journal of Agricultural and Food Chemistry, 49(9),


PT

4352-4358. doi: 10.1021/jf0013407


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Graphical Abstract
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Highlights

• Potential use of non-Saccharomyces yeasts in brewing processes.


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• Different metabolic traits in non-conventional yeasts for brewing.

• Influence of non-conventional yeasts on bioflavor, low alcohol and functional beers.

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