Food Research International 119 (2019) 378–389

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Food Research International

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Review

Lachancea yeast species: Origin, biochemical characteristics and oenological T

significance
Tristan Jade Porter, Benoit Divol, Mathabatha Evodia Setati
⁎

Institute for Wine Biotechnology, Department of Viticulture and Oenology, Stellenbosch University, Stellenbosch 7600, South Africa

ARTICLE INFO ABSTRACT

Keywords: The genus Lachancea, first proposed in 2003, currently comprises 12 valid species, all found to have eight
Lachancea chromosomes. Lachancea spp. occupy a myriad of natural and anthropic habitats, and their geographic as well as
β-Glucosidases ecological origin have been identified as key drivers in the genetic variations amongst strains of several of the
Taxonomy species. Lachancea thermotolerans is the type species of the genus and also the most widely explored, especially
Wine fermentation
for its role in fermentation environments. Indeed, L. thermotolerans is desired for its ability to acidify beer and
Enological traits
wine through the production of lactic acid, and to enhance aroma and flavor through increased production of
various compounds. Similarly, L. fermentati has been characterized for its potential contribution to the chemical
composition of these beverages, albeit to a lesser extent, while other species have received little attention.
Overall, members of the genus Lachancea form part of the microbiomes in many fermentation ecosystems and
contribute directly or indirectly to the modulation of aroma and flavor of different products. The current review
provides an overview of this genus, including the latest reports on the genetic and biochemical characteristics of
member species, as well as their biotechnological potential.

1. Introduction fermentation of various substrates, in particular grape must. This re-

Yeasts are routinely involved in many modern biotechnological
applications, including the production of metabolites and recombinant
proteins as well as in vivo biotransformations (Mattanovich, Sauer, &
Gasser, 2014). Most commonly, yeast species have been associated with
the fermentation of alcoholic beverages and food. Although Sacchar-
omyces cerevisiae has been acknowledged as the premier yeast species in
wine fermentation, in recent years attention has shifted towards the
potential of non-Saccharomyces yeasts (Ciani & Comitini, 2011; Padilla,
Gil, & Manzanares, 2016). Positive contributions from several yeast
species have been noted, (Cordero Otero et al., 2003; Gonz̕alez-Royo
et al., 2014), and this has subsequently led to the commercialization of
some strains, amongst them Lachancea thermotolerans (Belda et al.,
2016). Strains of L. thermotolerans are frequently used for the acid-
ification of low-acidity wines produced in warmer climate regions
(Benito, 2018). As a moderate fermenter, shown to express enzymes of
oenological interest and enhance positive flavor and aroma in wine, this
yeast species has gained considerable interest in research (Beckner
Whitener et al., 2015, 2016; Varela, 2016). Consequently, two recent
reviews have highlighted the wine-relevant features of L. thermotolerans
(Benito, 2018; Morata et al., 2018). However, the genus Lachancea is
diverse, currently comprising 11 valid species, most of which occur in
view offers a detailed analysis of the current status of the genus La-
chancea with focus on three major focus areas (i) taxonomic delineation
and ecological distribution (ii) biochemical traits, (iii) biotechnological
potential.
2. Historical overview of Lachancea species classification
The genus Lachancea was first proposed by Kurtzman in 2003, fol-
lowing a reclassification of several yeast genera based on genetic re-
latedness rather than phenotypic similarities. The genus now comprises
species that were previously classified in the genera Zygosaccharomyces,
Kluyveromyces and Saccharomyces. Historically, these species were
clustered into their respective genera based on morphology of vegeta-
tive cells and sexual states as well as fermentation and growth tests
common for yeast identification (Fell, Statzell-Tallman, & Kurtzman,
2004; Kurtzman & Robnett, 2003). The unreliability of such tests can be
highlighted by the reclassification of L. fermentati a total of six times.
Indeed, Zygosaccharomyces fermentati (Naganishi, 1928) was changed to
S. cerevisiae in 1952 (Lodder & Kregler-van Rij, 1952), back to Z. fer-
mentati in 1954 (Kudryavtsev, 1954), to Saccharomyces montanus in
1956 (Phaff, Miller, & Shifrine, 1956), to Torulaspora manchurica in
1975 (van der Walt & Johannsen, 1975), back to Z. fermentati again in

⁎
Corresponding author.
E-mail address: setati@sun.ac.za (M.E. Setati).

https://doi.org/10.1016/j.foodres.2019.02.003
Received 15 October 2018; Received in revised form 30 January 2019; Accepted 1 February 2019
Available online 04 February 2019
0963-9969/ © 2019 Elsevier Ltd. All rights reserved.
T.J. Porter et al.
1977 (von Arx, Rodrigues de Miranda, Smith, & Yarrow, 1977) and
finally to L. fermentati in 2003 (Kurtzman, 2003), as discussed below.
At the beginning of the 21st century, Kurtzman and Robnett (2003)
employed a multigene sequence analysis strategy to develop a dataset
for various yeasts comprising nucleotide sequences of unlinked genes;
18S rDNA, ITS1–5.8S rDNA-ITS2 and 26S rDNA, translation elongation
factor 1α (EF-1α), actin-1 and RNA polymerase II nuclear genes as well
as cytochrome oxidase II (COX II) mitochondrial genes. This dataset led
to the reclassification of various yeast species and the introduction of
the genus Lachancea (Kurtzman, 2003). Included into this proposed
novel genus, were the species Lachancea cidri (formerly Zygosacchar-
omyces cidri), Lachancea fermentati (Zygosaccharomyces fermentati), La-
chancea thermotolerans (Zygosaccharomyces thermotolerans and Kluyver-
omyces thermotolerans), Lachancea kluyveri (Saccharomyces kluyveri) and
Lachancea waltii (Kluyveromyces waltii). L. thermotolerans was selected as
the type species for this genus. Following the proposal of the Lachancea
genus by Kurtzman (2003), several other species including L. meyersii,
L. dasiensis, L. nothofagi, L. mirantina, L. lanzarotensis and L. quebecensis
were subsequently isolated, identified and placed within this genus. An
increase in the frequency of yeast species added into the Lachancea
genus can be noticed following its proposal in 2003 (Fig. 1), which
could be due to the increase in ease and reliability of species classifi-
cation, with the introduction of genetic sequencing.
Currently, the genus Lachancea has 11 valid species, which based on
D1/D2 sequence analysis, groups into 4 phylogenetic clusters (Fig. 2).
The first cluster comprises L. thermotolerans, L. quebecensis, L. waltii,
L. dasiensis, L. nothofagi, L. meyersii and L. lazarotensis, while the second
and third cluster contain, L. kluyveri, and L. fermentati and L. cidri, re-
spectively, while the fourth cluster comprises L. mirantina. Phylogenetic
analysis of the mitochondrial genes of 7 of the species, revealed similar
clusters (Friedrich, Jung, Hou, Neuvéglise, & Schacherer, 2012). Fur-
thermore, L. kluyverii has so far been highlighted as the most divergent
of the genus, based on the phylogenetic relationships of the mi-
tochondrial genomes (Friedrich et al., 2012).
Overall, several reports have shown that while showing high levels
of synteny in their protein coding regions, differentiation lies in the
intergenic regions and intron content of the Lachancea spp. and mi-
tochondrial genome sequence similarity can be linked to their geo-
graphical locations (Freel, Friedrich, Hou, & Schacherer, 2014;
Friedrich et al., 2012; Jung, Friedrich, Reisser, Hou, & Schacherer,
2012).
Z. thermotolerans S. cidri
Z. fermentati S. kluyveri
Fig. 1. Timeline depicting the initial taxonomic characterization, the eventual re-characterization of several Lachancea spp. (*) and the subsequent addition of
recently isolated yeast species to the Lachancea genus. (Yarrow, 1972)
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Food Research International 119 (2019) 378–389
3. Lachancea yeast species isolation and biochemical
characteristics
3.1. Natural habitats from which Lachancea spp. were isolated
Lachancea species have been isolated from a wide variety of ecolo-
gical niches, including plants, insects, soil, food and beverages (Fig. 3).
Amongst the species, L. thermotolerans and L. fermentati have been iso-
lated most frequently and from the widest diversity of ecological niches
(Fig. 3), most noticeably from grape must. In contrast, several La-
chancea spp. have been encountered less frequently, including L. da-
siensis, L. quebecensis, L. mirantina and L. lanzarotensis. Strains of several
species such as L. dasiensis (Lee, Yao, Liu, Hsieh, & Young, 2009),
L. nothofagi (Mestre, Ulloa, Rosa, Lachance, & Fontenla, 2010), L. fer-
mentati and L. quebecensis (Freel, Charron, Leducq, Landry, &
Schacherer, 2015) have been isolated from various parts of plants in-
cluding leaves, bark and fluxes. It has been proposed that these yeasts
could be associated with insects, perhaps drosophilids that frequently
visit sap fluxes and tree bark (Mestre et al., 2010). Indeed, this is
plausible since L. kluyveri and L. fermentati were previously isolated
from the intestinal canal of Drosophila wild species (Phaff et al., 1956).
There is a lack in literature, delving into why these Lachancea species
are able to survive in such a wide variety of ecological niches.
3.2. Biochemical traits of Lachancea spp.
Following the reclassification of several yeast species into the newly
proposed genus Lachancea.Fell et al. (2004) highlighted that members
of this genus possess certain common substrate assimilation and fer-
mentation characteristics. Other reports have also reported common
characteristics for the genus as a whole (Benito, 2018). However, with
the discovery of new Lachancea spp., these common characteristics no
longer hold true. As of now, research has shown that Lachancea species
can typically ferment glucose and at least one other sugar, as well as
assimilate raffinose, ethanol (with the exception of L. dasiensis, L. no-
thofagi and L. mirantina) and mannitol. They are also typically unable to
assimilate nitrate, lactose, soluble starch, L-, D-arabinose, D-ribose (ex-
cept L. quebecensis), L-rhamnose, D-glucosamine, N-acetyl-D-glucosamine
(except L. mirantina), methanol, erythritol, galactitol, citrate, inositol
and hexadecane (Table 1). A noticeable profile is that of L. walti, where
the strains analyzed have an inability to assimilate inulin, 2-keto-D-
gluconate, glycerol, D-galactose, maltose, trehalose, melezitose, succi-
nate, lactate, melibiose or ∂-Methyl‑D-glucoside as well as grow on
L. mirantina
L. nothofagi
L. meyersii
et al.
L. dasiensis
L. quebecensis
K. waltii
L. fermentatii
L. thermotolerans
K. thermotolerans L. kluyveri
L. cidri
L. waltii
L. lanzarontensis
T.J. Porter et al. Food Research International 119 (2019) 378–389

Fig. 2. The phylogenetic relationships of the Lachancea spp. was inferred using the Neighbour-Joining method and based on the 546 bp of the D1/D2 domain of the
26S rRNA. The tree is drawn to scale, with branch lengths (next to the branches) in the same units as those of the evolutionary distances used to infer the phylogenetic
tree. The evolutionary distances were computed using the Maximum Composite Likelihood method and the evolutionary analyses were conducted in MEGA7. The
outgroup species used was Kluyveromyces marxianus.

media supplemented with 10% NaCl. Lachancea spp. Trehalose protects S. cerevisiae cells when exposed to
lethal ethanol concentrations of over 10% (v/v) – a common environ-
ment of wine fermentations (Bandara, Fraser, Chambers, & Stanley,
4. Oenological potential of Lachancea spp.
2009). The intracellular uptake of ethanol during carbohydrate fer-
mentation places the yeast cells in a stressful environment. A me-
Amongst the members of the genus Lachancea, L. thermotolerans and
chanism of survival involves metabolic responses including the accu-
L. fermentati have been associated with grape must and wine fermen-
mulation of trehalose, likely through its membrane-stabilizing action
tation processes in several wine producing countries and over many
(Sebollela et al., 2004). Investigation into whether this is the metabolic
years, while L. lanzarotensis has only recently been isolated.
process followed by Lachancea spp. is therefore required, as well as
Consequently, more attention has been paid to the oenological potential
whether Lachancea spp. are able to synthesize trehalose, enabling their
of L. thermotolerans and to a lesser extent L. fermentati. Currently, only
survival in stressful conditions, such as a wine fermentation.
one report (Porter, Divol, & Setati, 2019) has investigated the oenolo-
The strains of the three wine-related yeast species have also been
gical potential of L. lanzarotensis (fermentation behavior and organo-
seen to grow on the 50% D-glucose supplemented medium (Table 1),
leptic contribution), utilizing only two strains. This yeast species is,
suggesting to their increased osmotic tolerance, which is an important
therefore, in need of additional focus in literature. For the purpose of
characteristic for yeast in wine fermentations. Grape must contains high
this review, the available information regarding the oenological po-
concentrations of sugars, or osmotically active substances, which can
tential of L. thermotolerans and L. fermentati, is discussed further.
lead to the cells entering hyperosmotic shock. Yeast cells adapt by cell
wall and cytoskeleton modification or the synthesis/uptake of an os-
4.1. Biochemical traits of oenological interest molyte, such as glycerol (Bauer & Pretorius, 2000). L. fermentati and
L. lanzarotensis strains displayed variability in glycerol assimilation
Observing the reported assimilation profiles (at least for the strains while various L. thermotolerans strains showed the ability to assimilate
analyzed) for the wine-related Lachancea spp. (Table 1), the data sug- this compound. Investigation of the yeast species behavior in wine
gest potential biochemical characteristics allowing adaptation to the fermentations, specifically for L. thermotolerans and L. fermentati has,
harsh winemaking conditions. All three of the yeast species have been however, provided more information on their adaptation capabilities
shown to assimilate trehalose. Unlike many of the carbon sources, and is discussed below.
utilized mainly for proliferation, trehalose metabolism has been linked
to stress resistance in yeast cells. Under stress, trehalose concentration
can reach as high as 35% of the dry weight of the cells (Wiemken,
1990). This link was however made utilizing S cerevisiae, and not

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T.J. Porter et al. Food Research International 119 (2019) 378–389

Fig. 3. Various habitats and ecological niches from which classified Lachancea spp. have previously been isolated; Lf: L. fermentati, Lt: L. thermotolerans; Lk: L. kluyveri,
Lc: L. cidri; Ld: L. dasiensis, Lme: L. meyersii, Ll: L. lanzarotensis, Lw: L. waltii, Ln: L. nothofagi, Lm: L. mirantina and Lq: L. quebecensis. The segment colors separate the
Lachancea spp., and the size of each segment represents the frequency each species has been isolated from a particular habitat. The references for the corresponding
reports are noted and common references illustrated by corresponding letters (Abegg et al., 2016; Aponte and Blaiotta, 2016a; Bagheri et al., 2015; Barata et al.,
2008; Barnett et al., 1983; Basílio et al., 2008; Biedunkiewicz et al., 2013; Clavijo et al., 2010; Coton et al., 2006; Couto et al., 2005; da CP Miguel et al., 2011; Esteve-
Zarzoso et al., 2001; Fernández et al., 1999; Filipov, 1932; González et al., 2013; González et al., 2007; Hsieh et al., 2012; Kodama and Kyono, 1974; Lachance et al.,
1988; Legakis, 1961; Magalhães et al., 2010; Magalhães et al., 2011; Mesquita et al., 2013; Morgan, 2016; Nisiotou et al., 2007; Nova et al., 2009; Osorio-Cadavid
et al., 2008; Pereira et al., 2011; Schwan and Wheals, 2004; Senses-Ergul et al., 2006; Shihata and Mrak, 1952; Tzanetakis et al., 1998; Wojtatowicz et al., 2001; Xufre
et al., 2006; Zheng et al., 2011; De Santo, Galego, Gonçalves, & Quintas, 2012; Marsh, O'Sullivan, Hill, Ross, & Cotter, 2014).

4.2. L. thermotolerans – behavior during fermentation of grape juice and
contribution to resulting wines
4.2.1. L. Thermotolerans population genetics
Recently Banilas, Sgouros, and Nisiotou (2016) performed genetic
and phenotypic typing of 47 L. thermotolerans isolates from two dif-
ferent regions in Greece. Using microsatellites PCR, the study showed
that L. thermotolerans isolates displayed distinct genetic clusters based
on geographic origin. Similarly, Hranilovic, Bely, Masneuf-Pomarede,
Jiranek, and Albertin (2017) demonstrated that L. thermotolerans strains
from anthropic and natural environments formed separate genetic
clusters, and that within these clusters, the strains grouped together
based on origin. Within the isolates from anthropic environment, two
major domestic groups mainly comprising isolates from grape and wine
related environments were identified, further revealing that the evo-
lution of L. thermotolerans was driven by geographical origin as well as

Table 1
Substrate assimilation profiles of different Lachancea species.
Assimilation profiles LF LT LK LC LW LM LD LN LMi LL LQ

L-Sorbose V + V + + W/− − − − − V
Inulin V V − V − V − − S − +
2-Keto-D-gluconate V V V − − V − V − nd V
Ethanol +/S V + + + S − − − +/W S
Glycerol V + V + − S − + + V +
D-Galactose +/V +/V + + − -/W + D S +/W +
Maltose +/W + + +/V − + + + S + +
Trehalose +/W + +/V + − + W + S + V
Melezitose + + +/V -/V − + + + − + +
Succinate +/V V +/V + − − − − − − nd
DL-Lactate +/W − +/V +/V − − − − S − +
Melibiose -/V -/V + + − − + − − − +
∂-Methyl‑D-glucoside + + + + − + nd nd nd nd nd
Growth on:
50% D-Glucose + + +/V +/V + -/V + V − + +
0.01% Cyclohexamide + − − + + − + − − − −
10% NaCl v/+/s -/V − + − + − V − + S

nd: no data available; D: Delayed growth; S: slow growth; V: variable growth; W: weak growth; −: negative; +: positive; LF: L. fermentati; LT: L. thermotolerans; LK:
L. kluyveri; LC: L. cidri; LW: L. waltii; LM: L. meyersii; LD: L. dasiensis; LN: L. nothofagi; LMi: L. mirantina; LL: L. lanzarotensis; LQ: L. quebecensis.

381
T.J. Porter et al.
the ecological niche of the isolation source. Moreover, the study by
Banilas et al. (2016) showed that the strains from different regions
displayed distinct oenological phenotypes. The oenological traits tested
included ethanol tolerance, SO2 resistance, H2S production, flocculation
phenotype, fermentation rate, tritable acidity and volatile acidity. In
addition, Hranilovic et al. (2018) also revealed distinct phenotypic
performance of L. thermotolerans genetic groups mainly based on fer-
mentation performance and production of volatile metabolites.
4.2.2. L. thermotolerans – fermentation kinetics
L. thermotolerans is the only Lachancea species currently commer-
cialized for use in the wine making industry. Chr. Hansen provides
three products containing L. thermotolerans, namely; Concerto™ – con-
sisting of L. thermotolerans alone, Melody™ – a combination of
L. thermotolerans, Torulaspora delbrueckii and S. cerevisiae, and Viniflora®
Rhythm – a blend of S. cerevisiae and (Kluyveromyces) L. thermotolerans.
These products are advertized to enhance aspects of wine including,
acidity, aroma complexity and intensity, fruitiness and mouth feel
(Benito, 2018; Du Plessis et al., 2017).
While a range of investigations have been performed to evaluate the
oenological impact of L. thermotolerans, these reports have been carried
out in differing grape matrices, fermentation conditions and utilizing
various yeast strains. Drawing conclusions from these studies regarding
the behavior of the L. thermotolerans species as a whole would therefore
be inaccurate. Indeed, several studies have indicated great variability in
fermentative phenotypes between different yeast strains, such as
Escribano et al. (2018). More specifically, this deep genotype diversity
was illustrated amongst L. thermotolerans isolates by Banilas et al.
(2016), which can explain differing results obtained when making use
of different L. thermotolerans strains in wine fermentations. Certain
trends, irrespective of the changing conditions, can however be noticed.
The single inoculation of L. thermotolerans into white and red grape
must has been shown to result in the presence of residual sugars and
lower ethanol concentrations, due to its lower fermentation rate
(Balikci, Tanguler, Jolly, & Erten, 2016; Ciani, Beco, & Comitini, 2006;
Cordero-Bueso, Esteve-Zarzoso, Cabellos, Gil-Díazm, & Arroyo, 2013;
Gobbi et al., 2013; Mora, Barbas, & Mulet, 1990; Mostert & Divol, 2014;
Porter et al., 2019). For the completion of the fermentation, co-culture
or sequential inoculation with S. cerevisiae is therefore necessary. Al-
ternatively, L. thermotolerans can be used in co-fermentation with other
strongly fermentative non-Saccharomyces yeasts such as Schizosacchar-
omyces pombe and Torulaspora delbrueckii (Benito, Calderón, Palomero,
& Benito, 2015; Escribano-Viana et al., 2018).
L. thermotolerans has been seen to persist till the middle and end
stages of the fermentation during mixed culture fermentations with
S. cerevisiae. Reported for both simultaneous and sequential inoculation
strategies, the consequence of inoculating L. thermotolerans is the re-
sulting lowered fermentation rate. Although, when adding
L. thermotolerans and S. cerevisiae simultaneously the cumulative CO2
loss is increasingly similar by the end of the fermentation. All si-
multaneous fermentations, irrespective of varying dosage, have conse-
quently resulted in similar ethanol and residual sugar levels in the wine
compared to their respective S. cerevisiae controls (Balikci et al., 2016;
Benito, Calderon, & Benito, 2016; Ciani et al., 2006; Comitini et al.,
2011; Gobbi et al., 2013; Kapsopoulou, Mourtzini, Anthoulas, &
Nerantzis, 2007; Porter et al., 2019). Sequential fermentations have, in
general, been shown to result in the increased persistence of L. ther-
motolerans in the mixed fermentations, where the later addition of
S. cerevisiae has allowed the L. thermotolerans yeast cells to reach higher
concentrations and subsequently illustrate increased competitiveness
(Ciani et al., 2006; Kapsopoulou et al., 2007; Gobbi et al., 2013; Balikci
et al., 2016; Benito, Calderon, & Benito, 2016,Benito, Calderón,
Palomero, & Benito, 2016; Beckner Whitener et al., 2017). Specifically
how long the L. thermotolerans is able to persist and dominate in the
fermentations has however varied, which was explained by
Kapsopoulou et al. (2007) as being dependent on the ability for
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Food Research International 119 (2019) 378–389
L. thermotolerans to reach a critical cell population, namely 7 log cfu/
mL, prior to the inoculation of S. cerevisiae. Upon critical analysis, while
this hypothesis was true in several reports (Balikci et al., 2016; Ciani
et al., 2006; Gobbi et al., 2013; Nurgel, Erten, Canbas, Cabaroglu, &
Selli, 2005), it has also been invalidated in others (Beckner Whitener
et al., 2016; Beckner Whitener et al., 2017; Benito, Calderon, & Benito,
2016). This, therefore, suggests the dominance and competitive nature
of L. thermotolerans, to perhaps be strain dependent. Indeed, Porter et al.
(2019) found two L. thermotolerans strains to behave differently, re-
garding their survival in a sequential fermentation with S. cerevisiae.
The death of this yeast species can also be attributed to various biotic
factors.
The decline in L. thermotolerans during fermentations has also been
attributed to the impact of parameters such as higher temperatures
(Balikci et al., 2016; Ciani et al., 2006; Gobbi et al., 2013), lack of
oxygenation (Holm Hansen, Nissen, Sommer, Nielsen, & Arnerborg,
2001; Shekhawat, Bauer, & Setati, 2017), cell-to-cell contact with
S. cerevisiae (Nissen & Arneborg, 2003; Nissen, Nielsen, & Arneborg,
2003) and the production of toxic compounds by S. cerevisiae
(Albergaria, Francisco, Gori, Arneborg, & Gírio, 2010). Therefore, fur-
ther investigation into the ability of various L. thermotolerans strains to
survive till the end of fermentations, in spite of these inhibitory factors,
could unveil interesting characteristics and provide insight into traits
which could further uncover additional commercially viable yeasts.
4.2.3. L. thermotolerans – impact on wine quality
Although L. thermotolerans does not consistently persist to the end of
the fermentation, it has still been determined to positively influence the
analytical and sensorial profile of wine. Various investigations have
been performed analyzing L. thermotolerans in co-culture fermentations
with S. cerevisiae. These investigations were however performed under
varying conditions have been carried out in diverse grape matrices,
utilizing different strains of both species (Table 2). The determined
metabolite concentrations in the resulting wines are thus different for
each report; however, certain trends are noticeable when comparing the
co-culture fermentation to those completed with S. cerevisiae alone. For
instance, co-culture fermentations with L. thermotolerans typically result
in higher titratable and total acidity. This is generally attributed to the
reduction in acetic acid and increase in D,L -lactic acid levels, respec-
tively (Mora et al., 1990; Ciani et al., 2006; Kapsopoulou et al., 2007;
Comitini et al., 2011; Gobbi et al., 2013; Balikci et al., 2016; Beckner
Whitener et al., 2016; Benito, Hofmann, et al., 2015, Benito, Calderon,
& Benito, 2016,Benito, Calderón, et al., 2016, 2018; Porter et al., 2019).
While these traits could be considered a species phenotype, it is clear
that the production levels of lactic acid can vary between strains and
can range from 1 to 9 g/L (Vilela, 2018). Indeed, Hranilovic et al.
(2018) demonstrated significant variation in lactic acid production
within two major domestic groups of L. thermotolerans. Similarly, only
some strains have been shown to consume significant amounts of acetic
acid in aerated fermentations (Vilela, 2018).
Glycerol, able to impart a sweet taste and impact mouth fullness
(Nieuwoudt, Prior, Pretorius, & Bauer, 2002), was also seen to increase
with L. thermotolerans inoculation (Kapsopoulou et al., 2007; Gobbi
et al., 2013; Benito, Hofmann, et al., 2015, Benito, Calderon, & Benito,
2016,Benito, Calderón, Palomero, et al. 2016). While frequent, a sig-
nificant increase in glycerol and reduction in acetic acid was not ob-
served in all the reports, suggestions a reliance on either strain, fer-
mentation condition or the biomass and persistence of L. thermotolerans
(Table 2). Prolonging the persistence of L. thermotolerans, generally
through the implementation of sequential inoculation strategies, has
resulted in the respective increase/decrease of these compounds
(Balikci et al., 2016; Beckner Whitener et al., 2017; Benito, Calderon, &
Benito, 2016; Comitini et al., 2011; Gobbi et al., 2013; Kapsopoulou
et al., 2007). In contrast to these observed trends, the production of
ethanol and acetaldehyde varied within the different reports (Balikci
et al., 2016; Benito et al., 2015; Benito, Calderón, et al., 2016; Benito,
T.J. Porter et al. Food Research International 119 (2019) 378–389

Table 2
Mono and mixed-culture fermentations of Lachancea thermotolerans (Lt) and Saccharomyces cerevisiae (Sc).
Yeast strain Inoculation strategy (day) Dosage (cfu/mL) Scale of Grape variety or media Reference
fermentation
Lt Sc Lt Sc

6
Unknown I Mono 10 / (100L) Unsterilized Manto Negro Mora et al. (1990)
grape must
M8 DBVPG 101 SIM 106 106 (200 mL) Paterurized Pinot grigio Ciani et al. (2006)
SEq. (4) grape must
TH941 SCM952 SIM 5 × 105 5 × 106 (200 mL) Sterile grape must Kapsopoulou et al.
SEq. (1) (2007)
SEq. (2)
SEq. (3)
101⁎⁎ EC1118 SIM 107 107 (105 (103 (500 mL) Pasteurized grape must Comitini et al. (2011)
CLI 1219 Mono 106 / (400 mL) Filter sterilized Malvar grape Cordero-Bueso et al.
must (2013)
101⁎⁎ / Mono 107 / (1 L) Pasteurized Sangiovese grape Gobbi et al. (2013)
EC1118 SIM 107 106 must
SEq. (1)
SEq. (2)
SIM Industrial scale
SEq. (2) (10hL)
IWBT 1326⁎⁎⁎ / Mono 2 × 107 / Small (600 mL) Synthetic grape juice Mostert and Divol
VIN13 SEq. (1) 2 × 105 (2014)
6
Concerto™ / Mono 10 / Small (55 mL) Syrah grape must Beckner Whitener et al.
/ / Sauvignon grape must (2015)
6
Concerto™ EC1118 SEq. (2) 10 107 (4 L) Sterilized Riesling grape must Benito, Calderón, et al.
(2015)
CBS 2860 / Mono 5 × 106 / (800 mL) Sterilized white grape must Balikci et al. (2016)
⁎From former SIM 5 × 106 5 × 106 cv. Emir
work SEq. (1)
SEq. (2)
SEq. (3)
Concerto™ Enoferm (M2 SEQ (Sc added when 2% 106 106 (10L) Sauvignon blanc grape must Beckner Whitener et al.
(Lallemand ethanol v/v/ reached) (2016)
CECT 12672 CECT 12512 SIM 2.95 × 107 1.18 × 107 (3.9 L) Sterilized Airén grape must Benito, Calderon, &
SEq. (4) Benito, 2016
Concerto™ Enoferm (M2, SEQ (Sc added when 2% 106 106 (10 L) Shiraz grape must (SO2 Beckner Whitener et al.
Lallemand ethanol v/v/ reached) addition for sterilization) (2017)

SIM = simultaneous; SEQ = sequential.

⁎
Strains not specified in report, noted as strain isolated in previous work.
⁎⁎
Strain from culture collection of the Dipartimento di Scienze della Vita e dell'Ambiente DiSVA of the Polytechnic University of Marche.
⁎⁎⁎
Strain from culture collection of Institute of Wine Biotechnology (IWBT), Stellenbosch University.

Calderon, & Benito, 2016; Ciani et al., 2006; Gobbi et al., 2013;
Kapsopoulou et al., 2007; Porter et al., 2019).
The production of various volatile compounds can significantly
impact the overall perception of wine. Critical analysis found 2-phe-
nylethanol to increase irrespective of the varying fermentation condi-
tions (Table 2), with a fold increase between 1.22 and 1.91 relative to
the S. cerevisiae monoculture fermentations (Beckner Whitener et al.,
2015; Benito, Calderon, & Benito, 2016; Comitini et al., 2011; Gobbi
et al., 2013; Porter et al., 2019). The concentrations of this compound
were reported above its sensory detection thresholds and would,
therefore, be able to impart flowery/pollen aromas to the wine
(Beckner Whitener et al., 2015). Regarding ester production, the only
trend common in the varying reports was the increase in ethyl lactate
(buttery, cream aroma) and ethyl hexanoate (green apple and anise
aroma) as well as the reduction in phenylethyl acetate (fruity aroma)
(Gobbi et al., 2013; Benito, Hofmann, et al., 2015, 2016, 2018; Porter
et al., 2019). Sensory analysis performed on the co-culture fermented
wines reported on the increased overall impression and total acidity
(Benito, Calderon, & Benito, 2016; Gobbi et al., 2013). Beckner
Whitener et al. (2016, 2017), also analyzing sensory perception, high-
lighted the role differing grape must could play in determining the wine
quality. L. thermotolerans inoculated Shiraz wines were determined as
more distinct than Sauvignon blanc wines, when compared to those
produced by S. cerevisiae alone. L. thermotolerans inoculation was seen
to enhance the production of 1-ethyl-1 h-pyrorole-2-carboxaldehyde,
which is beneficial to Shiraz wines due to imparting a smoky/roasted
aroma.
The combined use of L. thermotolerans and Sc. pombe in sequential
fermentation has also been shown to produce red wines with increased
color intensity and hue (Benito, Calderón, et al., 2016,Benito, Calderón,
& Benito, 2017; Escott et al., 2018). This ability to influence wine color
was attributed to the low anthocyanin adsorption capacity of L. ther-
motolerans (Benito et al., 2017), complemented by the increased pro-
duction of pyruvic acid, which condenses with malvidin to form stable
polymeric pigments and vitisins (Benito, Calderón, et al., 2016; Escott
et al., 2018), although the latter might be considerably negligible. Al-
ternatively, the increase in color may be enhanced due to higher acidity
in wine, which would influence mainly the monomeric anthocyanins.
Indeed, studies have reported that between 20 and 25% of this class of
anthocyanins exist in the flavylium ion state, which is the state that
expresses color at wine pH of 3.4–3.6. This level declines rapidly as the
pH rises, thus resulting in a loss in color (He et al., 2012). However,
since the sequential fermentations using L. thermotolerans and
Sc. pombe, produced wine with higher pH (pH 3.8) than for instance the
wine produced with S. cerevisiae alone (pH 3.6–3.7) or with S. cerevisiae
and L. thermotolerans (pH 3.6), we can infer that the low anthocyanin
adsorption by L. thermotolerans makes a stronger contribution than
monomeric anthocyanins.
Strains of L. thermotolerans have been screened for the production
and release of polysaccharides in mono- and mixed-culture fermenta-
tions (Domizio, Liu, Bisson, & Barile, 2014; Romani et al., 2010). The
strains were shown to produce a considerably higher amounts of total
polysaccharides continuously during growth and throughout fermen-
tation compared to S. cerevisiae. However, intraspecific variation was

383
T.J. Porter et al.
observed with regard to the capacity to release polysaccharides
(Domizio et al., 2014; Romani et al., 2010). Similar observations have
been made for L. fermentati, although only a couple of strains have been
tested for this species (Domizio et al., 2011; Romani et al., 2010).
Chemical characterization of the polysaccharides showed that they
largely comprised mannoproteins (Domizio et al., 2014). Mannopro-
teins are amongst the main components of the yeast cell wall; they have
been reported to contribute towards reduction in tartaric acid pre-
cipitation, protein haze and wine astringency (Juega, Nunez,
Carrascosa, & Martinez-Rodriguez, 2012; Domizio et al., 2014). Aging
on lees can improve the release of mannoproteins. However, Belda et al.
(2016) showed that some strains short aging on lees from some strains
of L. thermotolerans did not result in significant increase in mannopro-
tein content compared to S. cerevisiae control strains.
Overall, there seems to be more commonality in the production of
primary metabolites than that of secondary metabolites, where a higher
dependence on yeast strain and fermentation conditions is noticeable.
The investigations performed for L. thermotolerans (Table 2) have pro-
vided insight into the yeast species fermentation capabilities and po-
tential sensorial impact. However, because various fermentation con-
ditions or parameters are not kept constant between the different
studies, it is difficult to conclude what impact L. thermotolerans actually
has on wine quality. Future investigations, where single parameters are
changed and the impact on L. thermotolerans is evaluated, could provide
an opportunity where the optimal conditions for this species are de-
termined; to ultimately produce better quality and distinct wine pro-
ducts using L. thermotolerans inoculation.
4.3. L. fermentati – behavior during fermentation of grape juice and
contribution to resulting wines
Extensive investigation into L. fermentati as a potential wine yeast is
notably limited in literature, where few studies were performed in the
1990s and more recently in 2013 and 2018. L. fermentati was previously
thought to form part of the Zygosaccharomyces genus, and possess a
close relation to Z. bailii, a known wine spoilage yeast species (Romano
& Suzzi, 1993b). This affiliation with a spoilage microorganism could
explain the limited investigation regarding its potential positive con-
tribution to wine quality.
4.3.1. L. fermentati – fermentation kinetics
Currently, majority of reports have only investigated L. fermentati in
monoculture fermentations, with only one recent report providing in-
sight into the behavior of L. fermentati in co-culture fermentations
(Porter et al., 2019). Various reports have shown L. fermentati to possess
average to high fermentation vigor in monoculture fermentations
(Porter et al., 2019; Romano & Suzzi, 1993b; Romano, Suzzi, Domizio,
& Fatichenti, 1997). Cordero-Bueso et al. (2013) reported a
2.2 ± 0.7% of daily sugar consumption for L. fermentati, superior to
that of L. thermotolerans. The competitive nature of this species was il-
lustrated by Romano et al. (1997), where L. fermentati strains replaced
yeast species including Kloeckera apiculata, Candida stellata and Candida
valida during the middle stages of spontaneous fermentations and were
present at the end of the fermentation with S. cerevisiae. While this data
suggests L. fermentati to be a strong fermentative yeast, able to ferment
grape must and persist till the later stages of the fermentation, the re-
sulting residual sugars from these fermentations were not specified.
Only Cordero-Bueso et al. (2013) and Porter et al. (2019) reported on
the residual sugar levels of L. fermentati monoculture fermentations, and
while it was lower than other species analyzed, co-culture with S. cer-
evisiae would be necessary to complete the fermentation.
4.3.2. L. fermentati – impact on wine quality and potential biotechnological
advantage
Monoculture fermentations with L. fermentati have been reported to
result in low acetaldehyde, H2S and SO2 production as well as, in
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Food Research International 119 (2019) 378–389
comparison to S. cerevisiae, increased volatile acidity and lowered ti-
tratable acidity (Cordero-Bueso et al., 2013; Romano et al., 1997;
Romano & Suzzi, 1993b). The production of acetic acid does however
vary, whereby reduced acetic acid has been reported (Cordero-Bueso
et al., 2013; Romano et al., 1997; Romano & Suzzi, 1993b) as well as
significantly higher production (Porter et al., 2019). This suggests
acetic acid production by L. fermentati to be strain or condition depen-
dent. The production of various volatile compounds can also impact the
quality of wine. L. fermentati has very limited research into the pro-
duction of volatile compounds and critical analysis of the common
compounds suggests strain differentiation regarding the production of
higher alcohols, Isoamyl alcohol and propanol (Cordero-Bueso et al.,
2013; Porter et al., 2019; Romano et al., 1997; Romano, Fiore,
Paraggio, Caruso, & Capece, 2003; Romano & Suzzi, 1993a). The pro-
duction of isobutanol could potentially be medium dependent, where
low levels were reported when fermented in synthetic medium and
levels comparable to S. cerevisiae when fermenting grape must
(Cordero-Bueso et al., 2013; Romano et al., 1997; Romano et al., 2003;
Romano & Suzzi, 1993a). Indeed, Porter et al. (2019) found sig-
nificantly higher levels of isobutanol production when fermenting real
grape must. Increased investigations into the volatiles produced by
L. fermentati, during monoculture and co-culture fermentations, will
provide an opportunity to gage insight into what one may expect from
L. fermentati as a species.
Various L. fermentati strains have also been shown to flocculate
(Porter et al., 2019; Romano & Suzzi, 1993b; Suzzi, Romano, &
Benevelli, 1992). Flocculation of yeast cells often involve the binding of
flocculins (lectin-like proteins) on the cell walls of flocculent cells that
selectively bind to the mannose residues in adjacent yeast cell walls
(Verstrepen, Derdelinckx, Verachtert, & Delvaux, 2003). This behavior
can aid in the efficiency of grape must clarification and processing
(Rossouw, Bagheri, Setati, & Bauer, 2015). The various L. fermentati
strains analyzed by Romano and Suzzi (1993b), for their ability to
flocculate and their degree of flocculation, illustrated strain depen-
dence. The same four strains were determined to flocculate in synthetic
must as well, however to a lesser degree (poor to very flocculent).
L. fermentati flocculation was also determined to be highly proteinase-
and trypsin-resistant, however, an irreversible inhibitory effect on
flocculation was seen when mannose was present (Suzzi et al., 1992).
Taking work by Verstrepen et al. (2003) into account, this suggests that
the free mannose in the media was able to bind to and occupy the
flocculin receptors, thus preventing direct interaction with the mannose
of adjacent yeast cells and flocculation. The flocculating behavior of
several L. fermentati strains could potentially benefit wine production;
however, like the fermentation behavior and organoleptic contributions
of this yeast species, increased research is required.
4.4. Extracellular enzyme production
In addition to the production of various primary and secondary
metabolites, yeast species can also influence wine aroma and the
technological process of wine production, through the expression of
various extracellular enzymes. Several of these enzymes are produced
and transported to the periplasmic space and secreted to the extra-
cellular medium, where they are able to interact with grape-derived
aroma precursor compounds (Strauss, Jolly, Lambrechts, & van
Rensburg, 2001). Enzymes of oenological relevance, commonly in-
vestigated for wine yeasts, include protease, pectinase and glycosidases.
4.4.1. Protease and pectinase enzyme activity
Unstable protein, of grape origin, forming a haze is an issue often
faced during the bottling of white and rosé wines. While not noxious,
the appearance of protein haze in wine can result in consumer rejection.
It is, therefore, common for winemakers to utilize protein removal
techniques, such as bentonite fining. It has however been reported that
not all the proteins are adsorbed and utilization of bentonite can lead to
T.J. Porter et al.
Table 3
Summary of protease, polygalacturonase and β-glucosidase activity screenings.
Enzyme activity L. thermotolerans L. fermentati⁎
(+Ve) (-Ve) (+Ve) (-Ve)
activity activity activity activity
Protease – 100% – Indeed, aside from degrading anthocyanins, β-glucosidases are able
No. of strains 98
Polygalacturonase 2.02% 97.98% 9.09% 90.91%
No. of strains 97 11
β-Glucosidase 13.27% 86.73% 80% 20%
No. of strains 111 10
⁎
As far as could be determined, no L. fermentati strains have been screened
for protease and very few for polygalacturonase and β-glucosidase activities.
reduced wine volume (5–20%) and alteration of the wine sensory
characteristics, due to loss of aroma and flavor compounds (Lagace &
Bisson, 1990). An alternative method involves exploiting the yeasts
ability to express proteases. Proteases are able to hydrolyze proteins
into smaller and more soluble compounds, which can aid in wine
clarification and stabilization (Fernández, Ubeda, & Briones, 2000).
Another advantage of protein hydrolysis is the resulting increase in
amino acids and peptides, aiding in the prevention of stuck or sluggish
fermentations due to the resulting increase in assimilable nitrogen
(Fernández et al., 2000; Lagace & Bisson, 1990). Another enzyme with
oenological relevance is pectinase. Pectinolytic enzymes, through the
degradation of pectin, can aid in enhancing the efficiency of grape
pressing and clarification of the must as well as increasing substance
extraction, contributing to wine color and aroma (Fernández et al.,
2000).
As summarized in Table 3, of the various L. thermotolerans strains
analyzed, negative results were reported for 100 and 98.20% of these
strains for protease and polygalacturonase activities, respectively
(Sakai, Okushima, & Yoshitake, 1984; Schwan, Cooper, & Wheals,
1997; Romo-Sánchez, Alves-Baffi, Arévalo-Villena, Úbeda-Iranzo, &
Briones-Pérez, 2010; Alimardani-Theuil, Gainvors-Claisse, & Duchiron,
2011; Comitini et al., 2011; Cordero-Bueso et al., 2013; Mostert, 2013;
Belda et al., 2016; Porter et al., 2019). This, therefore, suggests most
L. thermotolerans strains to lack this enzyme activity. Varying meth-
odologies (e.g. substrates and pH) were however utilized to measure
this activity. Regarding the activities for L. fermentati, although also
illustrating strain dependence, drawing any form of conclusions from
the available data is difficult due to the overall limited number of
screenings performed. Nevertheless, expression of protease and poly-
galacturonase activity from various L. fermentati strains has been re-
ported as negative for 100 and 91.67% of the strains analyzed, re-
spectively (Cordero-Bueso et al., 2013; da Silva, Borges, Medina,
Piccoli, & Schwan, 2005; Porter et al., 2019; Romo-Sánchez et al.,
2010). It cannot be ignored that many of these investigations utilize
varying substrates and techniques, which can ultimately affect the in-
duction and/or intensity of the respective enzymes (Fernández-
González, Di Stefano, & Briones, 2003; Hernández, Espinosa,
Fernández-González, & Briones, 2003).
4.4.2. Glycoside hydrolase enzyme activity
The contribution of yeast species to wine quality has previously
been noted, specifically due to the production of sensorially significant,
yeast metabolism-derived, volatile compounds including esters, alco-
hols and acetates (Jolly, Varela, & Pretorius, 2014). However, a certain
proportion of varietal flavors of wines are due to the presence of grape-
derived volatile aromatic compounds (Cabaroglu, Selli, Canbas,
Lepoutre, & Günata, 2003). Monoterpenes, due to their pleasant floral
notes, play an essential role in the aroma profiles of wines, especially
that of white varieties, for example Muscat, Gewürztraminer and
Riesling. A large proportion of the varietal aroma compounds do
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Food Research International 119 (2019) 378–389
however appear glycosidically conjugated in young wines, resulting in
their inability to contribute to the aroma of wine, and therefore form a
large group of untapped aroma and flavor potential (Manzanares,
Ramón, & Querol, 1999; Manzanares, Rojas, Genovés, & Vallés, 2000).
These volatile compounds can be liberated from their precursors by
enzymatic hydrolysis.
to hydrolyze monoglucosides, resulting in the release of the mono-
terpenol and glucose. This enzyme does not, however, possess en-
doglucanase activity and therefore disaccharide glycosides, possessing
more than the one bound sugar moiety, cannot be hydrolyzed by only
β-glucosidases and requires additional action by other glycoside hy-
drolases. Glycoside hydrolases are involved in the sequential cleaving
of monoterpenol disaccharide precursors and involves the sequential
mode of action of several enzymes, depending on the bound sugar
molecule: α-L-arabinofuranosidase, α-L-rhamnopyranosidase or β-D-
apiofuranosidase and β-D-glucosidase (Günata, Bitteur, Brillouet,
Bayonove, & Cordonnier, 1988). The ability of the glycoside hydrolases
to execute the release of monoterpenols depends on their affinity for
grape-derived terpenoid aglycones and their activity under wine
making conditions, for example, low pH, fermentation temperatures,
high initial sugar and increasing ethanol levels. Although commercial
enzymatic mixtures are available, their composition is often unknown
and irregular (Roche, Coutinho, Delgadillo, Dias Cardoso, & Coimbra,
2005). For this reason, exploiting yeasts that exhibit glycosidase ac-
tivities could prove beneficial.
As summarized in Table 3, screenings of various L. thermotolerans
strains found only 12% of the strains to possess β-glucosidase activity,
whereas 88% lacked the activity. Forming a large percentage of these
results are the 88 L. thermotolerans strains analyzed by Belda et al.
(2016), of which a large portion illustrated negative activity. The re-
ports that investigated β-glucosidase activity were performed on plates,
where arbutin was utilized as the substrate for enzyme activity; and
although the pH was different for some, the methodology was similar.
Therefore, analysis of several L. thermotolerans strains illustrated
varying results; suggesting strain differentiation (Belda, Ruiz, et al.,
2016; Comitini et al., 2011; Cordero-Bueso et al., 2013; González-
Arenzana et al., 2016; Mostert, 2013; Porter et al., 2019; Romo-Sánchez
et al., 2010; Rosi, Vinella, & Domizio, 1994).
Like with the protease and polygalacturonase activity, few
L. fermentati strains have been analyzed for β-glucosidase activity. Of
the 11 strains screened, 72.73% and 27.27% were found to possess and
lack this activity, respectively (Cordero-Bueso et al., 2013; Porter et al.,
2019; Romo-Sánchez et al., 2010). As discussed above, plate screenings
cannot accurately represent what may occur in a wine fermentation,
due to differing conditions in which activity is tested and the consistent
use of artificial substrates that do not represent the natural substrates in
a wine fermentation. While β-glucosidase has been analyzed during
wine fermentations for other yeast species (Fia, Giovani, & Rosi, 2005),
only recently has this activity been investigated for Lachancea spp.
(Porter et al., 2019). Some Lachancea spp. were found to expressed
positive β-glucosidase activity under wine making conditions; the en-
zyme was found to be mainly cell wall associated and peaked in activity
with a corresponding peak in cell concentration (Porter et al., 2019.
This type of investigation into a wider variety of Lachancea spp. and
strains could provide more insight into whether additional Lachancea
spp. are capable of expressing this enzyme, and whether this is a
characteristic of the entire Lachancea genus or is strain dependent.
4.5. Antagonistic activity in Lachancea species
Over the past decade the use of many protection chemicals has been
curtailed in most wine producing countries, while the implementation
of Integrated Pest Management (IPM) systems has helped spur the
growth of research into biological control agents (BCAs; Medina et al.,
2017; Zhang, Apaliya, Mahunu, Chen, & Li, 2016). In particular,
T.J. Porter et al.
research into the potential of grape associated yeasts as alternatives to
control ochratoxigenic fungi, notably black aspergilli, has gained im-
petus. Such endeavors have led to the identification of strains of
L. thermotolerans that exhibited an ability to reduce the growth rate and
ochratoxin A (OTA) accumulation by Aspergillus section Nigri in vitro, on
detached grape berries as well as in greenhouse and field trials (Fiori
et al., 2014; Ponsone et al., 2016; Ponsone, Chiotta, Combina, Dalcero,
& Chulze, 2011). Varied modes of action including competition for
space and nutrients, regulating the transcription of ochratoxin polyke-
tide synthase encoding gene, and possibly the production of volatile
organic compounds (VOCs) that inhibit sporulation and growth have
been proposed (Chulze et al., 2015; Fiori et al., 2014; Zeidan, Ul-
Hassan, Al-Thani, Balmas, & Jaoua, 2018). However, the inhibitory
effects are dependent not only on the producing L. thermotolerans strain
but also on the ochratoxigenic fungal species, aw and temperature, and
the interaction between them (Ponsone et al., 2011, 2016). Further-
more, live and inactivated cells of L. thermotolerans were shown to re-
move upto 71% and 82% of ochratoxin A from buffer solution, sug-
gesting a high adsorption capacity which makes the strains valuable for
the detoxification of grape juice and wine fermentation (Zeidan et al.,
2018).
Beyond their inhibitory activities against filamentous fungi, certain
Lachancea species have also been reported to secrete killer toxins active
against other yeast species. Indeed, L. waltii (previously known as
Kluyveromyces waltii) was shown to have killer activity against
Sc. pombe, a yeast previously regarded as a wine spoilage agent (Kono &
Himeno, 1997). The latter authors also reported that L. thermotolerans
displayed killer activity against the strain of S. cerevisiae tested. Simi-
larly, a recent study by Aponte and Blaiotta (2016b) found strains of
L. thermotolerans that were antagonistic against S. cerevisiae. However, it
is safe to consider this trait as strain specific since previous work by
Comitini et al. (2011) and González-Arenzana et al. (2016) could not
detect this activity in several strains. Moreover, González-Arenzana
et al. (2016) found most of the L. thermotolerans strains to be neutral
towards S. cerevisiae killer activity.
5. Conclusion
Evaluating the Lachancea species has provided insightful informa-
tion on intra- and interspecies genetic diversity and the subsequent
effects of selective pressures guiding evolution. These yeast species
have also been isolated from extensively diverse habitats, suggesting to
their ability to adapt to these various environmental conditions even
when genetically similar. However, the majority of the Lachancea spp.
have received very little attention, only having been isolated and
characterized, and potential impactful information remains unexplored.
In enology, increasing focus has shifted onto non-Saccharomyces
yeasts and their potential usefulness to achieve various purposes. As of
now, only three Lachancea species have been associated with grape
berries and grape must – namely L. thermotolerans, L. fermentati and
most recently L. lanzarotensis. Even though these species have all been
isolated from the wine environment, research has specifically focused
on the fermentation behavior/potential of L. thermotolerans as well as its
expression of enologically relevant enzymes. In contrast, information is
lacking for L. fermentati, even though it has been found in this en-
vironment and shown the potential to possess robust fermentative
capabilities. L. fermentati additionally has very limited data available
regarding its expression of enzymes of oenological relevance. Majority
of the previous investigations into enzyme activity for L. thermotolerans
and L. fermentati have also been performed under conditions dissimilar
to that of a wine fermentation. The recent investigation into this ac-
tivity during wine fermentations is the first step towards understanding
the potential contribution Lachancea spp. has regarding enhancing the
organoleptic profile of resulting wine. This report is also the only one to
currently have investigated L. lanzarotensis. However, with increasing
analysis of a wide variety of strains, our understanding of this yeast spp.
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Food Research International 119 (2019) 378–389
as a potential wine yeast, can increase.
The potential impact Lachancea spp. have on the wine industry has
already been highlighted, however with the increasing number of
Lachancea spp. and relevant investigations to be done, more is yet to be
discovered.
Funding
This work was supported by the Wine Industry Network for
Expertise and Technology (Winetech)SU IWBT 16/02 and the National
Research Foundation-Technology and Human Resources for Industry
Programme (TP14080184824). Opinions expressed and conclusions
arrived at are those of the authors and are not necessarily to be at-
tributed to the funding agency.
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