E-mail: pe a .kneze[email p o ec ed]
Me abolic ea u es o Ca nobac e ium spp. ele an o ood
spoilage
Jelena CVETANOVIC, Pe a KNEZEVIC*
Uni e si y o No i Sad, Facul y o Sciences, Depa men o Biology and Ecology, T g Dosi eja Ob ado ica 2, 21 000
No i Sad, Se bia
Biologia Se bica, 2025, 47: 3-17
Re iew pape
DOI: 10.5281/zenodo.17678864
Summa y. Membe s o he genus Ca nobac e ium a e psych o ole an , acul a i ely anae obic lac ic acid bac e ia
equen ly isola ed om e ige a ed and acuum-packed ood p oduc s o animal o igin. This e iew explo es hei
me abolic and physiological ai s ha con ibu e o ood spoilage, wi h a pa icula ocus on C. mal a oma icum and C.
di e gens. These species a e me abolically e sa ile, capable o e men ing a ious ca bohyd a es and p oducing ola ile
o ganic compounds (VOCs) such as alcohols, ke ones, aldehydes, and biogenic amines, which a e s ongly associ-
a ed wi h undesi able senso y changes in mea , sea ood, and dai y p oduc s. Thei esilience unde s ess condi ions,
including cold empe a u es, ele a ed pH, and modi ied a mosphe e packaging, suppo s hei pe sis ence in ood
ma ices. P o eoly ic and lipoly ic ac i i ies u he enhance hei spoilage po en ial by deg ading p o eins and a s,
leading o o -odo and ex u al de e io a ion. A comp ehensi e unde s anding o he me abolic pa hways and spoilage
mechanisms o Ca nobac e ium spp. is essen ial o de eloping e ec i e ood p ese a ion s a egies and imp o ing
shel -li e managemen o pe ishable p oduc s.
Keywo ds: biogenic amines, modi ied a mosphe e packaging (MAP), psych o ole an bac e ia, e ige a ed s o age,
acuum-packaged oods, ola ile o ganic compounds (VOCs).
Accep ed: 11 Sep embe 2025 / Published online: 31 Oc obe 2025
INTRODUCTION
Ca nobac e ium spp. a e G am-posi i e, non-spo e-
o ming, acul a i ely anae obic ods, commonly ound in
e ige a ed and acuum-packed (VP) animal-based ood
p oduc s (Hammes and He el 2009). Genus Ca nobac e ium
belongs o he lac ic acid bac e ia (LAB) g oup ha consis s
o o e 60 gene a, and hose mos equen ly associa ed wi h
ood e men a ion besides Ca nobac e ium include Lac o-
bacillus, Lac ococcus, Leuconos oc, Pediococcus, S ep ococ-
cus, En e ococcus, and Weissella (Mokoena 2017). Al hough
ypically ca alase- and oxidase-nega i e, some s ains can
exhibi ca alase ac i i y in he p esence o heme (Ringø e al.
2002). They exhibi e men a i e me abolism, a e acul a-
i ely he e o e men a i e, and can me abolize a b oad ange
o ca bohyd a es such as glucose, uc ose, mannose, ibose,
and suc ose, wi h a iabili y among species (Kim e al. 2009).
Many s ains p oduce acids om suga s like ibose, ehalose,
and galac ose, and hyd olyze subs a es such as a ginine and
esculin, con ibu ing o hei me abolic e sa ili y (Hammes
and He el 2009). Du ing e men a ion, hey p edominan ly
p oduce L(+)-lac a e, while C. pleis ocenium also gene a es
e hanol, ace ic acid, and CO₂. These bac e ia a e psych o ol-
e an , able o g ow a o below 0 °C, and a e also halo ole an
(up o 8% NaCl) and alkaliphilic, ole a ing pH le els up o 9.5
(Cailliez-G imal e al. 2014; Piku a and Hoo e 2014). Such
en i onmen al adap abili y makes hem pa icula ly ele an
in he spoilage o oods s o ed unde e ige a ion and VP. C.
di e gens and C. mal a oma icum a e especially associa ed
wi h mea and ish spoilage, due o hei abili y o g ow unde
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Ј. C e ano ic e P. Kneze ic
anae obic, low- empe a u e, and mildly acidic- o-neu al pH
condi ions (5.4–6.1). Unde hese s o age condi ions, hey
become dominan membe s o he mic obio a and con ibu e
signi ican ly o senso y deg ada ion.
Compa a i e genomics e ealed a high gene ic di e si y
wi hin he genus Ca nobac e ium, wi h genome sizes anging
om 2.4 o 3.6 Mb. The p esence o genes in ol ed in bac-
e iocin p oduc ion, adap a ion o cold en i onmen s, and
u iliza ion o di e se ca bohyd a es unde lines he ecological
e sa ili y o hese species. Impo an ly, se e al Ca nobac e i-
um genomes ha bo genes encoding a V- ype ATPase, which
is belie ed o con ibu e o su i al unde cold and alkaline
condi ions, u he suppo ing hei dual ole as po en ial
biop o ec i e cul u es in ood and oppo unis ic pa hogens
in ish (Roh and Kim 2021).
Al hough ypically associa ed wi h ood, some s ains ha e
been epo ed o cause mixed in ec ions in s essed ish and
ha e also been isola ed om human clinical samples, including
pus and blood (Collins e al. 1987; Chmelař e al. 2002).
Thei me abolic ac i i y is linked o he p oduc ion
o ola ile o ganic compounds (VOCs) such as 2-me hyl-
p opanal, 3-me hylp opanal, 2-me hylp opanol, and 3-me h-
ylp opanol, which impa a mal y o spoiled a oma (Mille e
al. 1974). Addi ionally, Ca nobac e ium spp. show a s ong
co ela ion wi h TVB-N ( o al ola ile basic ni ogen) and
TBARS ( hioba bi u ic acid- eac i e subs ances), and a nega-
i e co ela ion wi h senso y indices like appea ance, ex u e,
and odo , u he highligh ing hei ole in ood spoilage.
Some species also show chi inoly ic ac i i y and he abili y o
u ilize inulin, s a ch, and glycogen, suppo ing hei adap a-
ion and pe sis ence in ood ma ices (Leisne e al. 2007;
Hammes and He el 2009).
The aim o his e iew is o summa ize and c i ically
discuss he key me abolic cha ac e is ics o Ca nobac e ium
species ha con ibu e o hei ole in he spoilage o e ig-
e a ed and VP ood p oduc s, wi h a ocus on hei e men-
a i e pa hways, p oduc ion o ola ile o ganic compounds
(VOCs) (Table 1), p o eoly ic and lipoly ic ac i i ies, and
en i onmen al adap abili y.
CARNOBACTERIUM AND FOOD SAFETY
Ca nobac e ium species ha e been widely de ec ed in a
a ie y o p ocessed mea and sea ood p oduc s, aising con-
ce ns abou hei ole in ood spoilage and con amina ion.
These lac ic acid bac e ia (LAB) a e equen ly associa ed wi h
spoilage o cooked and cu ed mea s, pa icula ly e minal
spoilage, and hei abili y o g ow a e ige a ion empe a u es
signi ican ly impac s ood sa e y (Samelis e al. 2006; Chenoll e
al. 2007). Despi e he use o ood addi i es like asco bic, lac ic,
ace ic, ci ic, so bic, and benzoic acids, as well as NaNO₂ and
smoke-de i ed phenols, Ca nobac e ium species con inue o
h i e in hese en i onmen s (Jø gensen e al. 2000; Dalgaa d
e al. 2003; Lakshmanan and Dalgaa d 2004).
Ca nobac e ium spp. p esence in mea p oduc s
Ca nobac e ium spp. ha e been widely de ec ed in p o-
cessed mea s, including bacon (Shaw and Ha ding 1984),
ham (Bo ch and Molin 1988; Jack e al. 1996), cooked poul-
y (Ba aka e al. 2000), p essu e- ea ed chicken (O’B ien
and Ma shall 1996), p ocessed po k and chicken (G an and
Pa e son 1991). No ably, C. mal a oma icum has also been
de ec ed in e men ed sausages (Schillinge and Lücke 1987;
La ou u e-Thi ey a and Mon el 2003), which con as s
wi h he ypical nonacidu ic en i onmen s associa ed wi h
ca nobac e ia (Hammes and He el 2006). Ca nobac e ium
mal a oma icum and C. di e gens a e equen ly isola ed
om mea p oduc s s o ed be ween −1.5 °C and 2 °C, domi-
na ing mic obial popula ions in bee , po k, lamb, and poul-
y. These species may comp ise up o 50% o G am-posi i e
o LAB isola es (McMullen and S iles 1993; Sakala e al. 2002;
Jones 2004). Thei p e alence is consis en ac oss VP, ae o-
bically s o ed, and modi ied-a mosphe e packaged (MAP)
mea s wi h CO₂/N₂ a ios anging om 10:90 o 80:20 (Shaw
and Ha ding 1984; G an and Pa e son 1991; McMullen and
S iles 1993; Ba aka e al. 2000; Sakala e al. 2002; Susiluo o
e al. 2003; Jones 2004; Bjö k o h 2005; Lau sen e al. 2005;
Viha ainen e al. 2007). O he species, including C. gallina-
ium (Tho nley 1957; Collins e al. 1987), C. mobile (Chenoll
e al. 2007), and C. i idans (Holley e al. 2002), ha e also
been isola ed om a a ie y o mea p oduc s.
Psych o ophic bac e ia like Ca nobac e ium spp. can
h i e a chilled empe a u es, especially du ing ca cass cu -
ing. The p ima y sou ce o con amina ion appea s o be
mea p ocessing en i onmen s, a he han he slaugh e ed
animals hemsel es. Unlike many o he spoilage bac e ia,
Ca nobac e ium spp. ha e no been isola ed om he gas-
oin es inal ac o skin o li es ock such as chicken, ca le,
pigs, o sheep, sugges ing ha p ocessing en i onmen s a e
he p ima y sou ce o con amina ion. Findings in he li e a-
u e a y ega ding he e ec i eness o p ocessing in elimi-
na ing Ca nobac e ium spp. Fo example, C. di e gens and
C. mal a oma icum de ec ed on b oile ca casses we e linked
o ai bo ne con amina ion wi hin p ocessing plan s a he
han he incoming poul y (Viha ainen e al. 2007). In con-
as , o he s udies ha e epo ed ha app op ia e p ocess-
ing p ocedu es can ma kedly educe o e en elimina e hese
species om aw ma e ials (Samelis e al. 1998), indica ing
ha he ou come depends s ongly on he speci ic p ocess-
ing condi ions and hygiene p ac ices employed. Molecula
echniques such as 16S RNA gene sequencing ha e deep-
ened ou unde s anding o Ca nobac e ium p esence along
he po k and poul y p oduc ion chains (Poi ie e al. 2020).
4 Biologia Se bica 47
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Iden ical ERIC-PCR p o iles o Ca nobac e ium de ec ed in
en i onmen al samples, sheep pel s, and lamb ca casses sug-
ges po en ial c oss-con amina ion ou es du ing p ocessing
(Mills e al. 2014). Pa ial 16S RNA gene sequencing has
e ealed he complexi y o aw bee mic obio a, wi h Ca -
nobac e ium p esen alongside Acine obac e , Bacillus, Jan-
hinobac e ium, Lac obacillus, Pseudomonas, Psych obac e ,
and S eno ophomonas (Aquilan i e al. 2016; Callahan e al.
2017; Choi e al. 2020; Yang e al. 2020). S udies on cooked
ham ha e shown ha species such as B. he mosphac a, C.
di e gens, C. mal a oma icum, L. ca nosum, L. gelidum, L.
mesen e oides, and S. p o eamaculans pe sis a he end o
p ocessing, wi h B ocho h ix, Ca nobac e ium, Lac obacillus,
Leuconos oc, Vib io, and Weissella domina ing a he end o
shel li e (Van Reckem e al. 2020).
P esence in dai y p oduc s
Raw milk con ains a di e se mic obial communi y ha
includes bo h bene icial bac e ia wi h echnological applica-
ions, such as lac ic acid bac e ia, and ha m ul spoilage o -
ganisms (E colini e al. 2009). The nu ien - ich composi ion
and nea -neu al pH o aw milk c ea e ideal condi ions o
mic obial p oli e a ion. Con amina ion can occu h ough
mul iple pa hways, including a m en i onmen s, animal
eed, bedding ma e ials, udde su aces, and milking equip-
men (Von Neubeck e al. 2015; Islam e al. 2018). Ini ially,
milk collec ed asep ically om hei e udde s con ains no
bac e ia, bu his changes as milking equency inc eases. The
bac e ial composi ion on ea skin e lec s he su ounding
a m en i onmen and can be in luenced by bedding ma e-
ials, which a e a ec ed by eeding and housing p ac ices.
Feed sou ces a y in hei mic obial con en depending on
pas u e condi ions, suppo ing he g ow h o bac e ia, yeas s,
and molds. Addi ionally, s able dus and inadequa e milking
hygiene con ibu e o mic obial con amina ion o ea su -
aces (F é in e al. 2018). The le el o bac e ial con amina ion
in aw cow milk is de e mined by se e al in e connec ed
ac o s: animal heal h and cleanliness, s able sani a ion, p e-
milking udde p epa a ion p ocedu es, milking echniques,
p ope cleaning and disin ec ion o milking equipmen and
s o age anks, app op ia e milk cooling and s o age p ac-
ices, and pe sonnel hygiene s anda ds (Cempí ko á 2007).
Dai y p oduc s, pa icula ly cheese, ep esen signi ican es-
e oi s o ca nobac e ia. Lac obacillus mal a omicus, la e
eclassi ied as Ca nobac e ium mal a oma icum, was ini-
ially desc ibed by Mille e al. (1974) ollowing i s isola ion
om milk samples cha ac e ized by a dis inc mal y a oma.
Among 30 F ench so - ipened and ed-smea cheeses made
om cow, ewe, o goa milk, ei he aw o pas eu ized, C.
mal a oma icum was ound in 10 samples, wi h isola es om
h ee o hem showing an i-Lis e ia ac i i y (Cailliez-G imal
e al. 2007). In hese cheeses, C. mal a oma icum was he p e-
dominan psych o ophic lac ic acid bac e ium. I was capa-
ble o g owing e en unde alkaline pH condi ions, eaching
high concen a ions (10⁸ o 10⁹ CFU/g) du ing subsequen
cold s o age a 4 °C (Cailliez-G imal e al. 2007). This species
may also ac as a po en ial ipening mic oo ganism in so
cheeses (Edima 2007).
P esence in sea ood
Ca nobac e ium spp. a e also eme ging as impo an
sea ood spoilage bac e ia (Ja ès e al. 2009; Noseda e al.
2012). They ha e been shown o domina e he mic obio a
o peeled b own sh imp s o ed a 4 °C by day 7 (Calliauw
e al. 2016) and apidly spoil cooked opical sh imp (Macé
e al. 2014). Ca nobac e ium mal a oma icum ep esen s a
signi ican spoilage mic oo ganism in sea ood p oduc s,
pa icula ly sh imp, wi h i s spoilage po en ial being highly
dependen on s o age condi ions and packaging me hods.
Ca nobac e ium mal oa oma icum demons a es he abili y
o p oduce o -odo compounds in sh imp (Dabadé e al.
2015) and gene a es ola ile compounds such as ammonia
and o he TVB-N compounds ha con ibu e o spoilage in
bo h cooked cold-wa e sh imp (Pandalus bo ealis) and op-
ical sh imp species (Lau sen e al. 2005; Ja ès e al. 2011;
Macé e al. 2014). The spoilage ac i i y o C. mal a oma icum
exhibi s empe a u e-dependen cha ac e is ics, wi h s ud-
ies showing ha while Pseudomonas psych ophila p oduces
signi ican ly highe amoun s o TVB-N and demons a es
as e g ow h a es a empe a u es anging om 0 o 15 °C,
C. mal a oma icum exhibi s supe io g ow h a es a ele a ed
empe a u es such as 28 °C. The compe i i e ad an age o C.
mal a oma icum becomes pa icula ly p onounced unde
oxygen-limi ed condi ions due o i s high esis ance o CO2
and anae obic en i onmen s (Al a o and
He nandez
2013),
allowing i o ou compe e ae obic spoilage bac e ia in MAP
o VP sys ems. Howe e , in ai -s o ed p oduc s, i s spoilage
impac is diminished in he p esence o mo e ac i e ae obic
spoile s such as Pseudomonas sp. This packaging-dependen
spoilage beha io has been demons a ed in compa a i e
s udies, whe e C. mal a oma icum showed educed spoilage
ac i i y compa ed o Pseudomonas agi in ai -s o ed mea
p oduc s (E colini e al. 2010; Casabu i e al. 2011), while
eme ging as he dominan spoile in modi ied a mosphe e
packaged cooked opical sh imp compa ed o o he bac e-
ial species (Macé e al. 2014). Bo h Lau sen e al. (2005) and
Noseda e al. (2012) concluded ha igh be o e he expi a-
ion da e o MAP sea ood, he p edominan mic oo ganisms
isola ed we e G am-posi i e bac e ia, speci ically C. mal a o-
ma icum, C. di e gens, and B. he mosphac a. A mul iplex
eal- ime quan i a i e PCR (qPCR) me hod was de eloped
and alida ed o he speci ic de ec ion and quan i ica ion o
Biologia Se bica 47 5
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Ј. C e ano ic e P. Kneze ic
C. mal a oma icum and C. di e gens in cold-smoked salmon.
This assay enables apid and accu a e moni o ing o hese
species in sea ood p oduc s, o e ing an e icien al e na i e
o con en ional cul u e-dependen echniques (Le Rhun e
al. 2021).
P esence in o he oods
In a s udy in es iga ing bac e ial eggshell pene a ion
and whole egg con amina ion, Ca nobac e ium sp. was
among se en bac e ial s ains es ed o hei abili y o in ade
and pe sis wi hin eggs. Using bo h an aga - illed egg model
and in ac eggs, Ca nobac e ium was shown o pene a e
eggshells and su i e wi hin whole eggs, achie ing a 17.5%
con amina ion a e, second only o Salmonella en e i idis (De
Reu e al. 2006). Ca nobac e ium di e gens MB421, isola ed
om e men ed g een oli es, exhibi ed s ong p obio ic cha -
ac e is ics, including ole ance o bile sal s (up o 1%), gas ic
juice (pH 2.2), and sal concen a ions up o 7%. I showed
an imic obial ac i i y agains se e al pa hogenic bac e ia and
was capable o e men ing a ious ca bohyd a es and p o-
ducing signi ican amoun s o o ganic acids. These p ope -
ies sugges i s po en ial use as a s a e cul u e in e men ed
oods and as a iable p obio ic candida e (Saeed e al. 2023).
In he s udy in es iga ing LAB in lou -based e men a ions,
Ca nobac e ium spp. was iden i ied among he isola es om
whea lou dough. These indings demons a e ha whea
lou suppo s a mo e di e se LAB communi y, including
Ca nobac e ium, which may con ibu e o he e men a ion
p ocess and a ec he dough’s biochemical p ope ies, such
as acidi ica ion and an imic obial ac i i y (Kleib e al. 2023).
ROLE OF CARNOBACTERIUM SPP. IN FOOD
SPOILAGE
Mea spoilage is a complex and mul i ace ed p ocess
d i en by a combina ion o chemical and biological ac o s,
esul ing in he de e io a ion o mea o a s a e un i o
human consump ion (G am e al. 2002). The nu ien - ich
composi ion o mea , i s a o able pH (5.5–6.5), and high
mois u e con en c ea e an ideal en i onmen o mic obial
g ow h. Spoilage is no solely he esul o lipid oxida ion o
enzyma ic eac ions bu is hea ily in luenced by mic obial
ac i i y (Nychas e al. 2007, 2008; Doulge aki e al. 2012).
Key con ibu o s o spoilage include mic obial species se-
lec ed unde speci ic en i onmen al and s o age condi ions.
Fac o s such as he animal’s physiological s a e a slaugh e ,
p ocessing, anspo a ion, p ese a ion, and s o age con-
di ions signi ican ly in luence he mic obial quali y o aw
mea (Nychas e al. 2008). Packaging and empe a u e a e
pa icula ly c i ical, e ige a ion a o s psych o ophic mi-
c oo ganisms, and he p esence o absence o oxygen in lu-
ences mic obial dynamics and spoilage po en ial (Nychas
e al. 2007, 2008; Doulge aki e al. 2012). A limi ed numbe
o mic obial species a e p ima ily esponsible o spoilage.
These include bo h G am-posi i e and G am-nega i e bac-
e ia, which, h ough ae obic o anae obic me abolism, p o-
duce spoilage compounds such as es e s, ke ones, aldehydes,
sul u compounds, amines, and ola ile a y acids (Dain y
e al. 1985; Lambe e al. 1991; Kakou i and Nychas 1994).
Among he dominan spoilage o ganisms a e LAB,
no ably Ca nobac e ium spp., which equen ly p e ail in
VP and MAP mea and sea ood (Leisne e al. 2007). LAB,
including Ca nobac e ium, a e o en in ol ed in he e mi-
nal spoilage o cooked and cu ed mea s (Samelis e al. 2006;
Chenoll e al. 2007). These bac e ia a e psych o ophic, ca-
pable o su i ing and p oli e a ing a low empe a u es, and
ha e been implica ed in he spoilage o e ige a ed chicken
unde ae obic condi ions (Liang e al. 2012), aw salmon
in MAP (Macé e al. 2013), and cooked sh imp, p oducing
o -odo s such as cheese/ ee , o e men ed smells (Ja ès e
al. 2011). In MAP poul y p oduc s, spoilage is la gely a -
ibu ed o G am-posi i e bac e ia like B. he mosphac a and
LAB, including Ca nobac e ium, Lac ococcus, Lac obacillus,
and Leuconos oc (Ba aka e al. 2000; Koo e al. 2005; Säde
e al. 2013). En i onmen al and packaging ac o s signi i-
can ly a ec mic obial selec ion: o ins ance, C. di e gens is
supp essed in MAP wi h high O₂ (20–40%) and 40% CO₂,
while i h i es unde ae obic s o age (E colini e al. 2011).
The g ow h o C. undi um is simila ly inhibi ed by oxygen
(F anzmann e al. 1991). Seasonal ac o s u he in luence
he p e alence o Ca nobac e ium. In wa m clima es, meso-
philic LAB such as Ca nobac e ium spp., L. sakei, L. cu a-
us, and Weissella spp. a e mo e equen ly associa ed wi h
spoilage (E colini e al. 2009, 2011; Doulge aki e al. 2010,
2012). Ca nobac e ium was ound o domina e bee samples
collec ed in July (Hwang e al. 2020).
B ocho h ix he mosphac a and Ca nobac e ium spp.
exhibi mode a e una ish spoilage po en ial, cha ac e ized
by he p oduc ion o bu e /ca amel-like odo s (Silbande e
al. 2018). Some C. mal a oma icum s ains ha e been impli-
ca ed in spoilage phenomena, such as ex u al deg ada ion
o salmon ille s (Mo zel e al. 1997). The exp ession o spoil-
age ai s is u he in luenced by en i onmen al ac o s. Fo
ins ance, essen ial oils ha e exhibi ed an imic obial e ec s
agains se e al spoilage-associa ed species, including L. sakei,
L. cu a us, C. mal a oma icum, B. he mosphac a, P. luo es-
cens, and Se a ia lique aciens (Oua a a e al. 1997). No ably,
o egano essen ial oil has been shown o modula e glucose
me abolism pa hways in lac ic acid bac e ia, he eby al e ing
he p oduc ion o spoilage- ela ed me aboli es (Nychas e al.
1998; Skandamis and Nychas 2001).
Among dai y p oduc s, su ace mold- ipened so
cheeses made om unpas eu ized milk, such as B ie, ha e
6 Biologia Se bica 47
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
been shown o suppo subs an ial mic obial g ow h. The
ela i ely high su ace pH o B ie cheese (pH 6.8–7.6) a-
cili a es he p oli e a ion o hese bac e ia, wi h popula ions
eaching concen a ions o 10⁸ o 10⁹ CFU/g. Fu he mo e,
C. di e gens and C. mal a oma icum a e p ominen mem-
be s o he LAB communi y p esen in he cu d o mozza-
ella cheese p oduced using unpas eu ized milk (Mo ea e
al. 1999). Lipoly ic o p o eoly ic ac i i y is obse ed a low
empe a u es in se e al gene a p esen in mino abundance,
including A h obac e spp. and Ca nobac e ium spp. In a
s udy by Vi hanage e al. (2014) examining he ex acellula
p o ease ac i i y o psych o ophic bac e ial popula ions in
aw milk, app oxima ely hal o he bac e ial isola es dem-
ons a ed p o eoly ic capabili ies. Isola es o Ca nobac e ium
consis en ly p oduced bio ilms a 10 °C o 4 °C making hem
he mos p e alen bac e ia among high o al bac e ial coun
aw milk samples and his con i ms ele a ed bac e ial coun s
can esul om bio ilms and poo hygiene in he milk p o-
cessing en i onmen (Hahne e al. 2019).
Ca nobac e ium spp. a e widely s udied o hei po-
en ial as biop o ec i e cul u es, alued o hei abili y o
enhance shel li e wi hou nega i ely impac ing senso y
quali ies. Among hem, C. mal a oma icum s ains KOPRI
25789 and MMF-32 ha e demons a ed no able an imic o-
bial ac i i y, p ima ily h ough bac e iocin p oduc ion, e ec-
i ely inhibi ing Lis e ia monocy ogenes and a ious G am-
nega i e bac e ia in cold-smoked salmon (Da bandi e al.
2021). Howe e , an imic obial e icacy is no solely depen-
den on bac e iocin syn hesis, as bo h bac e iocin-p oducing
and non-p oducing s ains ha e shown inhibi o y capabili-
ies (Nilsson e al. 1999). Bac e ia o his genus exhibi h ee
key biop o ec i e mechanisms: compe i ion, exclusion, and
displacemen . Thei abili y o p oduce an imic obial sub-
s ances, sec e e enzymes, and exp ess adhesion p o eins on
he cell su ace enables ca nobac e ia o inhibi pa hogenic
and oppo unis ic bac e ia. Genomic ea u es a e cen al o
his ac i i y, as an ex ensi e sec e ome and p o eome equip
ca nobac e ia o quickly colonize niches and ou compe e
o he mic oo ganisms (Leisne e al. 2007).
Ca nobac e ium species exhibi an imic obial p ope ies
agains a ious oodbo ne pa hogens and spoilage mic oo -
ganisms, including Lis e ia (Danielski e al. 2022), Salmonella
(Hu e al. 2019), Se a ia spp., Pseudomonas spp. (Zhang e
al. 2019), Campylobac e spp. (Smialek e al. 2018), Pseudo-
monas luo escens, B ocho h ix he mosphac a (Ca ala i e
al. 2019), and En e obac e iaceae (Imazaki e al. 2014) ac oss
sea ood, mea , and dai y applica ions. These cha ac e is ics
demons a e Ca nobac e ium’s capaci y o enhance ood sa e-
y measu es (Kou soumanis e al. 2021). Addi ionally, Ca no-
bac e ium shows p omise o use in plan -based ood sys ems
(Ag iopoulou e al. 2020), and plan -based ood i ems (Gee -
ae s e al. 2020). Resea ch conduc ed by González-G age a
e al. (2024) highligh s he signi ican p omise o in eg a ing
piscicolin CM22, a newly iden i ied bac e iocin p oduced by
a cold- ole an C. mal a oma icum s ain, in o edible coa -
ings (ECs) made om chi osan and ish gela in o imp o e
he quali y and sa e y o ish p oduc s. The esul s show ha
hese ECs no ably enhanced mic obiological p ope ies and
p olonged he s o age li e o esh ish main ained a bo h 4
°C and 8 °C empe a u es. Fu he mo e, chi osan-de i ed
ECs p o ided pH s abiliza ion and imp o ed colo p ese a-
ion, hus main aining he senso y and unc ional cha ac e -
is ics o he ish p oduc s.
METABOLIC PROCESSES AND PRODUCTS
OF CARNOBACTERIUM SPP. DURING MEAT
SPOILAGE
Me abolic p ocesses
In con as o he ex ensi e me abolic knowledge accu-
mula ed o lac ococci and lac obacilli in ecen yea s, me-
abolism o Ca nobac e ium spp. emains ela i ely unde -
s udied. These bac e ia demons a e s ong ae obic g ow h
capabili ies, wi h C. mal a oma icus showing a 10- old in-
c ease in g ow h yield when heme is supplemen ed o ae obi
-
cally cul u ed cells (Meisel e al. 1994). Unde hese condi-
ions, unc ional cy och omes o bo h b and d ypes become
de ec able, simila o he cy och ome induc ion obse ed
in lac ococci and en e ococci (Sijpes eijn 1970; Ri chey and
Seely 1976). The p esence o cydAB genes encoding cy o-
ch ome oxidase (bd) in Lac ococcus lac is (Gaudu e al. 2002)
sugges s ha when heme is accessible, C. mal a oma icum
possesses espi a o y me abolic po en ial and displays ca a-
lase ac i i y. This ca alase p ope y is sha ed among all Ca -
nobac e ium species (Ringø e al. 2002), wi h he excep ion
o C. pleis ocenium, which has no been examined o his
cha ac e is ic. Respi a o y me abolism co ela es wi h en-
hanced oxygen consump ion, dec eased lac a e p oduc ion,
and ele a ed ace oin and CO2 gene a ion.
Unde s anding gene exp ession unde di e en a mo-
sphe ic condi ions p o ides insigh in o mic obial me abolic
adap a ions. Al hough no signi ican di e en ial exp ession
o genes ela ed o gene al me abolic pa hways was de ec ed
by me a ansc ip omic analysis unde MAP condi ions, in-
cluding CO₂/O₂ and CO₂/N₂ en i onmen s, speci ic s ess
esponses we e obse ed. Fo ins ance, genes encoding su-
pe oxide dismu ase and o ganic hyd ogen pe oxide esis-
ance p o eins we e up egula ed unde CO₂/O₂ condi ions,
sugges ing a cellula esponse o oxida i e s ess (Madaman-
chi and Runge 2007; Li e al. 2013). Despi e he lack o sig-
ni ican ansc ip omic changes, me abolic ac i i y emained
e iden ac oss bo h a mosphe ic en i onmen s. The mos
Biologia Se bica 47 7
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Ј. C e ano ic e P. Kneze ic
abundan ly exp essed p o eins belonged o enzymes in ol ed
in glycolysis, indica ing a s ong glycoly ic lux. P e ious e-
sea ch has shown ha C. di e gens is homo e men a i e, as
i lacks phosphoke olase, an essen ial enzyme o he e olac ic
e men a ion (Höll e al. 2020). Ne e heless, ansc ip omic
da a indica e ha py u a e-de i ed me abolic byp oduc s
such as e hanol ( ia e hanol dehyd ogenase), ace a e ( ia
ace a e kinase), o ma e ( ia o ma e ace yl ans e ase), and
CO₂ ( ia py u a e dehyd ogenase) a e p oduced unde bo h
CO₂/O₂ and CO₂/N₂ condi ions. This sugges s a me abolic
lexibili y, wi h he p oduc ion o compounds ypically asso-
cia ed wi h he e o e men a i e pa hways e en in he absence
o phosphoke olase. P io s udies ha e indica ed ha g ow h
condi ions and py u a e a ailabili y in luence me aboli e o -
ma ion (Ba owsky and Henschke 2004), and ae obic condi-
ions in pa icula ha e been shown o enhance hese me a-
bolic p ocesses, no ably in C. mal a oma icum (Casabu i e
al. 2011). Ca nobac e ia exhibi acul a i ely he e o e men a-
i e ca bohyd a e me abolism, e men ing bo h hexoses and
pen oses. Excluding C. pleis ocenium, hexoses a e con e ed
o L(+)-lac a e, while also p oducing CO2, ace a e, and e ha-
nol. Fu he mo e, o ma e is p oduced unde anae obic con-
di ions (Bo ch and Molin 1989) and ace oin unde ae obic
condi ions. Pen ose me abolism yields L(+)-lac a e, ace a e,
and e hanol (Holzap el and Ge be 1983). Glucose p ocess-
ing occu s h ough he glycoly ic pa hway (De B uyn e al.
1987, 1988). In G oup I species, app oxima ely 75% o lac-
a e p oduc ion is de i ed om glucose, wi h less han 10%
o igina ing om o ma e and ace a e, while he emaining
p oduc s p esumably s em om endogenous subs a es in
nu ien - ich media (Hammes and He el 2009). G oup II
species show dis inc pa e ns, wi h C. pleis ocenium p oduc-
ing e hanol, ace a e, and CO
2
wi hou lac a e o ma ion om
ca bohyd a e e men a ion. This me abolic p o ile excludes
his species om he physiological de ini ion o lac ic acid
bac e ia, sugges ing me abolism esembling he Embden-
Meye ho pa hway in yeas . Simila ly, C. undi um and C. al-
e undi um do no gene a e lac a e om glyce ol, p oducing
exclusi ely o ma e, e hanol, and ace a e. Ce ain species o
s ains demons a e polysaccha ide hyd olysis and e men a-
ion capabili ies. Inulin u iliza ion occu s in C. di e gens, C.
gallina um, and speci ic C. mal a oma icum s ains. Amy-
loly ic ac i i y is documen ed h ough s a ch u iliza ion in
C. pleis ocenium and glycogen u iliza ion in C. mobile and
C. gallina um. The p esence o s a ch in plan ma e ials and
subsequen ly in ish in es inal ac s may e lec adap a ion o
Ca nobac e ium o ha ecological niche (Hammes and He -
el 2009). C. mal a oma icum u ilizes wo dis inc pa hways
o lac ose and galac ose me abolism: he Taga ose-6-phos-
pha e (Taga ose-6P) pa hway and he Leloi pa hway. Some
s ains possess genes encoding one o bo h pa hways, while
o he s lack hese genes en i ely. The Taga ose-6P pa hway is
ypically encoded by he lac gene clus e (e.g., lacA-G), and
is in ol ed in he phosphoenolpy u a e phospho ans e ase
sys em (PTS)-media ed up ake and phospho yla ion o lac-
ose o galac ose, ollowed by enzyma ic deg ada ion. The
Leloi pa hway, on he o he hand, includes genes like galK,
galT, galE, and galM, which sequen ially con e galac ose
in o glucose-6-phospha e (Iskanda e al. 2016). Genomic
compa isons ac oss mul iple C. mal a oma icum s ains
showed ha hese me abolic genes a e a iably loca ed on
ch omosomes and megaplasmids, and o en exhibi signs
o ho izon al gene ans e , duplica ion, and genomic ea -
angemen . In pa icula , he C. mal oa oma icum exhibi s
he highes in aspeci ic he e ogenei y among he LAB ana-
lyzed, a inding ha co ela es wi h i s p e iously epo ed
high gene ic di e si y (Rahman e al. 2014). These indings
sugges ha he e olu iona y adap a ion o lac ose me ab
-
olism in C. mal a oma icum has been shaped by en i on-
men al p essu es, pa icula ly in dai y en i onmen s, whe e
lac ose is a p ima y ca bon sou ce (Iskanda e al. 2016).
These bac e ia also possess he abili y o deg ade p o-
eins. Genomic analysis has e ealed ha C. mal a oma icum
and C. di e gens possess conse ed oligopep ide anspo e
sys ems (OppABCDF, D pT) and in acellula pep idases,
enabling hem o e icien ly impo and deg ade pep ides
de i ed om ood p o eins (Dallagnol e al. 2021). Expe i-
men al s udies on C. mal a oma icum s ains g owing in
aw ish p o ein ex ac s showed signi ican deg ada ion o
soluble p o eins (up o ~137 mg/100 mL) and accumula ion
o ee amino acids, con i ming hei ac i e p o eolysis du -
ing spoilage. This enhances hei capaci y o gene a e ee
amino acids ha can be u he con e ed in o spoilage me-
aboli es. Ca nobac e ia exhibi ed mode a e deg ada ion o
ish sa coplasmic p o eins (≤73%), leading o he elease o
small pep ides and ee amino acids, wi h alanine, glycine,
aspa agine, and a ginine showing he g ea es inc eases
in concen a ion (Dallagnol e al. 2021). Recen genomic
analyses ha e e ealed ha C. mal a oma icum and C. di-
e gens possess homologs o P B, a cell en elope-associa ed
p o ease (CEP) commonly associa ed wi h dai y lac ic acid
bac e ia such as Lac obacillus delb ueckii subsp. bulga icus
(Iskanda e al. 2017). These p o eases enable he ex acel-
lula deg ada ion o p o eins in o pep ides, which a e sub-
sequen ly in e nalized ia oligopep ide anspo sys ems
(OppABCDF and D pT) and hyd olyzed u he by in acel-
lula pep idases. This p o eoly ic sys em appea s o be con-
se ed in C. mal a oma icum and C. di e gens, bu no in
all Ca nobac e ium s ains, sugges ing a selec i e ad an age
in p o ein- ich en i onmen s such as mea , ish, and dai y
p oduc s. The abili y o exploi en i onmen al p o eins o
amino acid acquisi ion likely con ibu es o hei success in
8 Biologia Se bica 47
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
e ige a ed oods and plays a c i ical ole in spoilage h ough
he elease o ola ile ni ogenous compounds, o -odo s,
and deg ada ion o ex u e and senso y quali y. A ginine
hyd olysis p o ides an addi ional sou ce o adenosine i-
phospha e (ATP), and mos membe s o he genus Ca no-
bac e ium a e capable o i , hough i is absen in C. i idans
and C. mal a oma icum (Lau sen e al. 2005). In addi ion,
genes associa ed wi h he a ginine deaminase pa hway a e
exp essed. This pa hway con ibu es o ene gy gene a ion ia
subs a e-le el phospho yla ion and plays a ole in mic oen-
i onmen al pH egula ion, u he suppo ing adap a ion o
a ying en i onmen al condi ions (Höll e al. 2020). These
indings emphasize he me abolic e sa ili y o Ca nobac e-
ium spp., especially C. di e gens, unde MAP, and highligh
how speci ic me abolic ou es help sus ain cellula unc ion
and adap a ion unde s ess.
Ca nobac e ia also p oduce lipases, which a e consid-
e ed seconda y spoilage ac o s. This ac i i y has been ob-
se ed on oli e oil/Tween-80 aga a bo h 20 °C and 4 °C,
indica ing hei abili y o b eak down a s, gene a e la o
compounds, and con ibu e o ancidi y (Casabu i e al.
2011). Also, i has been epo ed ha hey a e able o hyd o-
lyze ibu y in, indica ing lipoly ic ac i i y (Papon and Talon
1988). Ca nobac e ium di e gens and C. mal a oma icum
did no inhibi he oxida ion o linoleic acid du ing g ow h
(Talon e al. 2000), sugges ing ha hese species a e unlikely
o p o ec mea p oduc s om quali y de e io a ion caused
by lipid oxida ion.
Ca nobac e ium spp. do no p oduce DNases o RN-
ases and no nuclease ac i i y has been associa ed wi h hem
(Pale ich e al. 2024). Some species, such as C. i idans and
C. di e gens, p oduce hyd ogen pe oxide, which eac s wi h
myoglobin in VP cooked cu ed Bologna sausage, causing
an undesi able g een discolo a ion (Bo ch and Molin 1989;
Holley e al. 2002; Pei son e al. 2003). Despi e his, he spe-
ci ic enzymes esponsible o hyd ogen pe oxide p oduc ion
emain uniden i ied (Remenan e al. 2015) (Fig. 1).
Me abolic p oduc s
F esh aw mea is cha ac e ized by a dis inc i e senso y
p o ile, including a bloody, me allic, and sal y as e and an
a oma simila o blood se um (Joo and Kim 2011). These
a ibu es a e la gely d i en by VOCs, which con ibu e sig-
ni ican ly o mea a oma due o hei high apo p essu es a
oom empe a u e (Ca e 2010). VOCs encompass a b oad
ange o chemical classes— a y acids, hyd oca bons, alde-
hydes, ke ones, alcohols, es e s, benzenoids, e penoids, and
a ious ni ogen- and sul u -con aining compounds (Es é ez
e al. 2003; Ri as-Cañedo e al. 2009). In addi ion o being
key con ibu o s o a oma, some VOCs can se e as species-
speci ic ma ke s in mea (Rødbo en e al. 2004; Wood e al.
2004; Ro a and Schiebe le 2006). Senso y e alua ions ha e
u he expanded he ol ac o y desc ip o s associa ed wi h
aw mea (E colini e al. 2010; Casabu i e al. 2011; La S o ia
e al. 2012). Ca nobac e ium spp. a e known con ibu o s o
VOC p oduc ion du ing chilled s o age unde ae obic, VP, o
MAP condi ions. These bac e ia syn hesize alcohols ia me -
abolic ou es including amino acid ca abolism, p o eolysis,
aldehyde educ ion om lipid oxida ion, and me hyl ke one
Fig. 1. Me abolic p ocesses and p oduc s du ing ood spoilage by Ca nobac e ium spp. (TBARS – hioba bi u ic acid- eac i e subs ances;
VOCs – ola ile o ganic compounds: TVB-N – o al ola ile basic ni ogen).
Biologia Se bica 47 9
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Ј. C e ano ic e P. Kneze ic
Table 1. Vola ile o ganic compounds p oduced by Ca nobac e ium spp.
Vola ile o ganic
compound (VOC)
Cha ac e is ic smell Common Food
Sou ces
Re e ences
Diace yl
(2,3-bu anedione)
Bu e y, popco n-like,
c eamy
Bu e , cheese, spoiled
mea / ish
Jo aud e al.
2001
Ace oin (3-hyd oxy-2-
bu anone)
Mildly bu e y, c eamy Dai y, spoiled mea Dain y e al. 1989
2,3-Pen anedione Ca amel-like Smoked o cooked
mea
Odeyemi e al.
2018
Bu anoic acid Rancid bu e , swea y,
sou milk
Spoiled dai y o mea Jones 2004
1-Oc en-3-ol Mush oom-like, ea hy Mea s, dai y, spoiled
ish
Leisne e al. 1995
2-me hylp opanal Sha p, bi e , sligh ly
ui y (aldehydic)
Bee (as an o - la o ),
e men ed dai y, aged
cheese, oxidized a s
Mille e al. 1974
3-me hylp opanalSwee , nu y, ui y,
sligh ly cheesy
Fe men ed oods,
ipened cheese, cu ed
mea s, wine
Mille e al. 1974
2-me hylp opanol Sha p, alcoholic, mildly
ui y
Bee , wine, e men ed
ui s, sou dough
Mille e al. 1974
3-me hylp opanolCheesy, ui y, alcoholic Aged cheese,
e men ed soy, bee ,
sou milk
Mille e al. 1974
3-me hylbu anal Mal y, nu y, cheesy,
ui y
Aged cheese, e -
men ed mea s, bee ,
chocola e
Ramí ez and
Ca a 2007
hexanal
G een, g assy, a y,
ancid in high concen-
a ions
F esh mea s, dai y,
oxidized a s, eg-
e ables
Ramí ez and
Ca a 2007
10 Biologia Se bica 47
Me abolic ea u es o Ca nobac e ium spp. ele an o ood spoilage
Vola ile o ganic
compound (VOC)
Cha ac e is ic smell Common Food
Sou ces
Re e ences
nonanal Flo al, ci us-like, a y Cooked mea s, dai y
p oduc s, ui s, eg-
e able oils
Ramí ez and
Ca a 2007
hep anal Fa y, oily, g een, ci us Animal a s, dai y,
oas ed mea , nu s
Ramí ez and
Ca a 2007
benzaldehyde Almond-like, nu y,
swee
Almonds, che ies,
ap ico ke nels, some
dai y p oduc s
Mu iel e al. 2004
2-pen anoneSwee , ui y, sol en -
like
Cooked mea , cheese,
some ui s
Tan e al. 2020
2-hep anone F ui y, banana-like, blue
cheese
Blue cheese, ipened
cheeses, cooked mea s,
some ui s
Tan e al. 2020
isopen yl alcohol Alcoholic, banana, uel
oil
Fe men ed be e ages,
cheeses, soy sauce,
o e ipe ui s
Tan e al. 2020
e hyl p opanoa eF ui y, pineapple, um Pineapple,
s awbe ies,
e men ed dai y,
baked goods
Tan e al. 2020
ime hylamine Fishy, ammonia-like Spoiled ish, sea ood,
some cheeses
P abhaka e al.
2021
dime hylamine Fishy, ammoniacal,
unpleasan
Spoiled mea s, ish,
e men ed p o ein-
ich oods, aged
cheeses, mea ex ac s
P abhaka e al.
2021
Biologia Se bica 47 11
