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Review
Abstract The principal pathways for the formation of flavour compounds in cheese (glycolysis,
lipolysis and proteolysis) are reviewed. Depending on variety, microflora and ripening conditions, lac-
tate may be metabolized by a number of pathways to various compounds which contribute to cheese
flavour or off-flavours. Citrate metabolism by citrate-positive lactococci or Leuconostoc spp. is
important in certain varieties (e.g., Dutch cheeses). Lipolysis results directly in the formation of
flavour compounds by liberating free fatty acids (FFA). FFA may also be metabolized to alkan-2-ones
and fatty acid lactones. Proteolysis of the caseins to a range of small- and intermediate-sized peptides
and free amino acids (FAA) probably only contributes to the background flavour of most cheese
varieties, but FAA are important precursors for a range of poorly-understood catabolic reactions
which produce volatile compounds essential for flavour.
cheese / flavour / volatile flavour compound
Table I. Some flavour compounds which have been identified in Cheddar cheese (adapted from
Urbach [138]).
Tableau I. Composs aromatiques identifis dans le fromage Cheddar (daprs Urbach [138]).
295
Figure 1. Principaux mcanismes biochimiques de laffinage : (a) protolyse, (b) lipolyse et (c) mtabolisme du lactose, du lactate et du citrate.
296 P.L.H. McSweeney, M.J. Sousa
Figure 2. General
pathways for lactate
metabolism in cheese.
Figure 2. Principales
voies mtaboliques du
lactate dans les fro-
mages.
Lactate can be oxidized in vitro to acetate and oxygen permeability of the rind or pack-
and CO2 by components of the non-starter aging material influence the availability of
lactic acid bacteria (NSLAB) present in hard O2 in the cheese. However, it is unlikely
cheeses ([53]; Fig. 3). The size of the cheese that this pathway occurs to a significant
Figure 3. Metabolism
of lactate by lactococci
(modified from [53]).
Figure 3. Mtabolisme
du lactate par les lacto-
coques (daprs [53]).
298 P.L.H. McSweeney, M.J. Sousa
extent in cheese due to its low redox poten- exhausted, P. camemberti metabolizes
tial (ca. 250 mV). Acetate, an important amino acids released from the caseins with
flavour compound in many cheeses, in addi- the production of NH3 [56]. Deacidification
tion to being formed from lactose by lactic of the cheese surface has a major impact on
acid bacteria (LAB) may also be formed as the growth of coryneform bacteria (Br. linens
a result of citrate and lactate metabolism does not grow below pH 5.8), plasmin activ-
(see below), or as a product of the catabolism ity (which is optimum at alkaline pH) and
of amino acids. has a major effect on cheese texture since
Propionibacterium sp. in Swiss-type Ca2+ precipitates at the surface as Ca3(PO4)2,
cheeses grow once the cheeses are trans- causing a Ca2+ gradient to develop within
ferred into the warm-room after brining and the cheese. Similar deacidification occurs
preferentially metabolize L-lactate to pro- in smear-ripened cheeses by metabolism of
pionate, acetate and CO2 (see Scheme 2). lactate by yeasts and moulds which initially
The carbon dioxide produced is essential dominate the surface microflora. Deacid-
for eye development; and propionate and, ification is essential for the growth of
to a lesser extent, acetate contribute to the coryneform bacteria, which dominate the
flavour of these cheeses. Fermentation of smear during ripening and are responsible
lactose and lactate in Swiss-type cheeses for the characteristic flavour of these cheeses
has been described by Steffen et al. [132] [93, 120].
and Turner et al. [137].
Milk contains ca. 8 mmol.L1 citrate,
Late gas blowing and off-flavours in cer- most of which is lost in the whey during
tain hard cheeses result from the metabolism cheesemaking, since ca. 94% of the citrate is
of lactate (or glucose) by Clostridium sp. to in the soluble phase of the milk. Neverthe-
butyric acid and H2 ([53]; Fig. 4). These less, the low concentration of citrate in
defects may be avoided by good hygiene, cheese curd (10 mmol . kg 1 ) is of great
addition of NO 3 or lysozyme, or by the importance since it may be metabolized to a
physical removal of spores by bactofuga- number of volatile flavour compounds by
tion or microfiltration. certain mesophilic starters (citrate-positive,
Metabolism of lactate is most extensive in Cit+, lactococci and Leuconostoc sp.) by
surface mould-ripened cheeses, e.g., pathways summarized in Figure 5. Citrate is
Camembert and Brie [67, 81, 106]. The not metabolized by S. thermophilus nor by
mesophilic starter bacteria produce lactic thermophilic lactobacilli, but is metabolized
acid in the curd (ca. 1%), which is quickly by certain mesophilic lactobacilli in the
metabolized by secondary microorganisms NSLAB flora. Citrate metabolism has been
(initially Geotrichum candidum and Debary- reviewed by several researchers [25, 26, 51,
omyces hansenii, followed later by Penicil- 61]. Cit+ microorganisms do not utilize cit-
lium camemberti and perhaps coryneform rate as an energy source, but rather it is co-
bacteria like Brevibacterium linens). The metabolized with lactose or some other
yeasts and moulds rapidly metabolize lactate sugar. The gene encoding the citrate trans-
to CO2 and H2O, and the pH of the cheese port mechanism is plasmid-encoded. The
surface increases. When the lactate has been principal flavour compounds produced on
Scheme 2.
Biochemical pathways during cheese ripening
Figure 4. Pathway
for the formation of
butyrate and H 2
from glucose or lac-
tate by Clostridium
sp.; Fd: ferredoxin
(modified from
[53]).
Figure 4. Produc-
tion de butyrate et
de H 2 partir de
glucose ou de lac-
tate par Clostridium
sp. ; Fd : ferredoxine
299
(daprs [53]).
300 P.L.H. McSweeney, M.J. Sousa
metabolism of citrate are acetate, diacetyl, mentation can cause undesirable openness
acetoin and 2,3-butanediol. Diacetyl is usu- and the floating curd defect in Cheddar
ally produced only in small amounts (110 and Cottage cheeses. Diacetyl is an impor-
gmL1 in milk), but acetoin is generally tant aroma compound in a number of vari-
produced in much higher quantities (1050- eties, including Dutch-type cheeses, Quarg
fold higher than diacetyl concentrations). and Cottage cheese. Diacetyl can be con-
Acetate is produced from citrate in equimo- verted to acetoin and 2,3-butanediol and
lar concentrations. Production of 2,3-butane- 2-butanone, which are also important flavour
diol by starters has not been studied in detail. compounds in some cheese varieties [30].
Despite its importance, the exact reactions
which result in the formation of diacetyl
remain unclear. Diacetyl could be produced 1.2. Lipolysis and metabolism
directly from acetaldehyde-thiamine pyro- of fatty acids
phosphate (TPP) and acetyl-CoA by
enzymic action, but diacetyl synthase has Milk fat is essential for the development
never been identified clearly in LAB. Alter- of the correct flavour in cheese during ripen-
natively, -acetolactate, formed from the ing. Cheddar and other cheeses normally
reaction of pyruvate and acetaldehyde-TPP, made from whole milk do not develop cor-
is very unstable and could be decarboxy- rect flavour when made from skim milk or
lated non-oxidatively to form acetoin, or milks in which milk fat has been replaced
oxidatively to form diacetyl. However, since with other lipids [43, 107, 147]. Indeed, sat-
the oxidation-reduction potential of cheese isfactory flavour development is one of the
(ca. 250 mV) is low, it is not clear how principal problems encountered in the man-
diacetyl could be produced oxidatively by ufacture of reduced-fat variants of estab-
starter cultures. Acetoin is produced from lished cheese varieties.
-acetolactate by the action of acetolactate As in all high-fat foods, lipids present in
decarboxylase. Products of citrate meta- cheese can undergo oxidative or hydrolytic
bolism produced by pure cultures of Cit+ degradation. Because of the negative oxi-
lactococci and Leuconostoc sp. differ: the dation-reduction potential of cheese, oxi-
former produce diacetyl, acetoin and CO2 dation of cheese lipids is probably limited;
in addition to lactate, but the latter produce but the extent to which it occurs, and its
large amounts of lactate and acetate. Pure contribution (if any) to cheese flavour devel-
cultures of Leuconostoc sp. do not produce opment has received little attention [53].
acetoin or diacetyl because pyruvate is an Enzymatic hydrolysis of triglycerides to
intermediate of both lactose and citrate fatty acids and glycerol, mono- or diglyc-
metabolism and the pyruvate produced from erides (lipolysis) is, however, essential to
citrate metabolism is converted to lactate flavour development in many cheese vari-
and acetylphosphate (via acetyl-CoA). eties. Milk fat contains high concentrations
Acetate is produced from acetylphosphate of short- and intermediate-chain fatty acids
with the concomitant production of 1 mol which, when liberated by lipolysis, con-
ATP, resulting in faster growth of the tribute directly to cheese flavour. The pro-
microorganism. In mixed cultures, Leu- portions of free C6:0 to C18:3 fatty acids in
conostoc sp. produce diacetyl and acetoin, Cheddar cheese appear to be similar to those
perhaps because their ability to take up lac- in milk fat, but free butyric acid (C4:0) occurs
tose is greatly reduced below pH 5.5. at a greater relative concentration in cheese
Citrate metabolism is of particular impor- than in milk fat, suggesting that butyrate is
tance in Dutch-type cheeses where the CO2 either selectively liberated by lipases pre-
produced is responsible for eye formation. sent in Cheddar or that it is synthesized by
Carbon dioxide produced by citrate fer- the cheese microflora [12]. The specificity of
302 P.L.H. McSweeney, M.J. Sousa
the lipase also influences the development of for very long periods (e.g., Grana-type
cheese flavour, since short-chain fatty acids cheeses) also develop quite high concen-
(which have the greatest flavour impact) are trations of FFA.
generally found at the sn-3 position of
triglycerides. Cheese pH also influences the Milk contains an indigenous lipase (lipo-
flavour impact of FFA, since carboxylic protein lipase, LPL) in addition to a number
acids and their salts are perceived differ- of esterases. LPL is the principal indig-
ently. enous lipase in milk, and has been well char-
acterized (see [108]). It is relatively non-
Lipolysis is particularly extensive in hard specific and liberates fatty acids from the
Italian varieties, surface bacterially-ripened sn-1 and sn-3 positions of mono-, di- or
(smear) cheeses and blue mould cheeses triglycerides and from the sn-1 position of
(Tab. II), and is essential to correct flavour phospholipids. The majority (> 80%) of LPL
development in these cheeses. Extensive in bovine milk is associated with the casein
lipolysis is considered undesirable in many micelle and thus is incorporated into the
internal, bacterially-ripened varieties such reticulum of the curd. LPL is most important
as Cheddar, Gouda and Swiss cheeses; high in raw milk cheeses since its activity is
levels of fatty acids in these cheeses lead to reduced by pasteurization, although it prob-
rancidity. However, low concentrations of ably also makes some contribution to lipol-
FFA contribute to the flavour of these ysis in pasteurized-milk cheeses since
cheeses, particularly when they are correctly 78 C 10 s is required for its complete
balanced with the products of proteolysis inactivation.
or other reactions [15, 53, 122].
Rennet extracts used in the production
Lipases in cheese originate from 6 sour- of most cheese varieties should be free from
ces: the milk, rennet preparation (rennet lipase activity, but rennet pastes used to
paste), starter, adjunct starter, non-starter coagulate the milk for certain Italian vari-
bacteria and, if used, exogenous lipases. The eties (e.g., Provolone, Romano) contain pre-
origin of lipases in varieties characterized gastric esterase (PGE) which is responsible
by extensive lipolysis is usually from the for the extensive lipolysis in these cheeses
coagulant (rennet paste in certain Italian (see [99]). PGE is secreted by glands at the
varieties), or from the adjunct starter (mould- base of the tongue and is washed into the
ripened cheeses). Cheeses which are ripened abomasum with the food. Rennet paste is
Table II. Typical concentrations of free fatty acids (FFA) in some cheese varieties [150, 151].
Tableau II. Concentration en acides gras libres selon le type de fromage [150, 151].
prepared from partially-dried abomasa by fied and characterized [116]. Penicillium sp.
grinding them into a paste. Rennet pastes produce potent extracellular lipases which
are added to cheese milk as slurry (in water are primarily responsible for the extensive
or milk). PGE is a 49 kgmol1 glycopro- lipolysis in mould-ripened cheeses (see
tein which is highly specific for short-chain [56]). P. camemberti excretes a lipase that is
fatty acids at the sn-3 position. Because of optimally active at about pH 9.0 and 35 C,
hygienic concerns regarding rennet pastes, while P. roqueforti excretes 2 lipases with
fungal lipases have been investigated as different substrate specificities that are opti-
alternatives (see [45, 48]). mally active between pH 6.0 and 6.5 and at
more alkaline pH values (7.59.5), respec-
The lipase/esterase systems of starter bac- tively. The impact of FFA on the flavour of
teria have received much less attention then blue mould-ripened cheeses is less than for
their proteolytic systems. Lactococcus sp. hard Italian varieties, possibly due to neu-
are only weakly lipolytic, but lactococci tralization as the pH increases during ripen-
may be responsible for the liberation of quite ing, and because of the dominant influence
high levels of FFA when present in high of methyl ketones on the flavour of blue
cell numbers or over extended ripening peri- mould cheeses.
ods. Lipases/esterases of Lactococcus
strains, which appear to be intracellular, In addition to their direct impact on
have been studied [22, 50, 53, 60]. Obli- cheese flavour, FFA also act as precursor
gately homofermentative lactobacilli used molecules for a series of catabolic reactions
as starters (Lb. helveticus, Lb. delbrueckii which lead to the production of other flavour
subsp. bulgaricus and Lb. delbrueckii subsp. compounds (Fig. 6).
lactis) also produce esterases, some of which The flavour of blue mould cheeses is dom-
have been studied (e.g., [37, 68]). Faculta- inated by alkan-2-ones (2-methyl ketones;
tively heterofermentative lactobacilli (e.g., see Scheme 3). A homologous series of
Lb. casei, Lb. paracasei and Lb. plantarum), alkan-2-ones with odd-numbered carbon
which dominate the NSLAB flora of many chains from C3 to C15 (principally heptan-2-
cheese varieties, are weakly lipolytic. Micro- one, nonan-2-one and undecan-2-one) is
coccus and Pediococcus are also weakly produced by the catabolism of FFA by
lipolytic [13]. Psychrotrophic bacteria (e.g., P. roqueforti.
Pseudomonas sp.) produce heat-stable
lipases which adsorb onto the fat globules in
milk and survive pasteurization. They may
contribute to lipolysis in cheese made from
milk containing high numbers of psy-
chrotrophic bacteria prior to pasteurization
[28]. Scheme 3.
Figure 7. Catabolism of
fatty acids by Penicillium
roqueforti (modified from
[53]).
Figure 7. Catabolisme des
acides gras par Penicillium
roqueforti (daprs [53]).
Biochemical pathways during cheese ripening 305
Scheme 4.
mould-ripened varieties); (ii) direct contri- the Phe105-Met106 bond of the micelle-sta-
bution to flavour and perhaps to off-flavour bilizing protein, -casein, during the coag-
(e.g., bitterness) of cheese through the for- ulation of milk. Most of the coagulant activ-
mation of peptides and free amino acids, ity added to the milk is lost in the whey, but
FAA; (iii) liberation of substrates (amino ca. 6% is retained in the curd depending on
acids) for secondary catabolic changes (e.g., factors including coagulant type, cooking
deamination, decarboxylation, transamina- temperature and pH at drainage; residual
tion, desulphuration, catabolism of aromatic coagulant contributes to proteolysis in many
compounds such as phenylalanine, tyrosine, varieties [29]. In high-cooked cheese (e.g.,
tryptophan and reactions of amino acids Emmental), chymosin is denatured exten-
with other compounds); and (iv) changes to sively and makes relatively little contribution
the cheese matrix, which facilitate the to ripening.
release of sapid compounds during masti- The dominant indigenous milk pro-
cation. teinase, plasmin (fibrinolysin, EC 3.4.21.7),
During ripening, proteolysis in cheese is has been the subject of much study (see [9,
catalyzed by enzymes from: (i) the coagulant 58] for reviews). The plasmin system in
(e.g., chymosin, pepsin, or plant or fungal milk is complex and consists of the active
acid proteinases); (ii) the milk (plasmin, enzyme (plasmin), its zymogen (plasmino-
cathepsin D and perhaps other somatic cell gen), plasminogen activators, and inhibitors
proteinases); (iii) the starter; (iv) the non- of plasmin (Fig. 8), all of which are present
starter; or (v) the secondary starter (e.g., in milk; plasmin, plasminogen and plas-
P. camemberti, P. roqueforti, Propionibac- minogen activators are associated with the
terium spp., Br. linens and other coryne- casein micelles in milk, while plasmin
forms); and (vi) exogenous proteinases inhibitors are in the serum phase. Plasmin is
and/or peptidases used to accelerate ripen- a trypsin-like serine proteinase which is opti-
ing. mally active at about pH 7.5 and 37 C, and
is highly specific for peptide bonds on the
In most cheese varieties, the initial
C-terminal side of lysyl, and to a lesser
hydrolysis of caseins is caused by the coag-
extent, arginyl residues [145]. It is particu-
ulant and to a lesser extent by plasmin and
larly active on s2- and -caseins; hydroly-
perhaps somatic cell proteinases (e.g.,
sis of the latter leads to the formation of
cathepsin D), which results in the forma-
-caseins (C-terminal fragments) and pro-
tion of large (water-insoluble) and interme-
teose-peptones (N-terminal fragments) [39].
diate-sized (water-soluble) peptides which
The specificity of plasmin on -casein [35],
are subsequently degraded by the coagulant
s2-casein [75, 143] and s1-casein [76, 90]
and enzymes from the starter and non-starter
in solution has been reported. Milk also con-
flora of the cheese. The production of small
tains an acid proteinase, now known to be
peptides and FAA is caused by the action
cathepsin D (EC 3.4.23.5), which is
of microbial proteinases and peptidases.
relatively heat-labile (inactivated at 70 C
The coagulants used to clot milk are 10 min), and has a pH optimum of 4.0
crude preparations of selected proteinases [66]. The specificity of cathepsin D is sim-
which often possess a considerable prote- ilar to that of chymosin [92] with prefer-
olytic activity. Chymosin (EC 3.4.23.4) is ence for s1-casein but, surprisingly, this
the major proteinase in traditional animal enzyme has very poor milk clotting activity
rennets (8894% milk clotting activity, [92]. In rennet free-cheeses, formation of
MCA), with the remainder being pepsin (EC s1-CN(f24-199) has been attributed to the
3.4.23.1; 612% MCA) [121]. The principal activity of the acid milk proteinase [142].
role of chymosin (or other coagulants) in In addition to cathepsin D, other proteolytic
cheesemaking is to specifically hydrolyze enzymes from somatic cells may also
Biochemical pathways during cheese ripening 307
(a)
(b)
Figure 9. (a) The action of the cell envelope-associated proteinase (PrtP) of Lactococcus on the
casein; and (b) degradation of a hypothetical tetrapeptide by the combined action of lactococcal
proteinases and peptidases (modified from [53]). PepO: oligoendopeptidase; PepI: prolyl iminopep-
tidase; PCP: pyrrolidone carboxylyl peptidase; PepN, PepC and PepG: general aminopeptidases;
PepL: leucyl aminopeptidase; PepA: glutamyl aminopeptidase; PepX: X-prolyl-dipeptidyl aminopep-
tidase; PepR: prolinase; PepQ: prolidase; PepV: dipeptidase; and PepT: tripeptidase.
Figure 9. (a) Action de la protase membranaire (PrtP) de Lactococcus sur les casines et
(b) mcanisme de dgradation dun ttrapeptide hypothtique par laction combine de protases
et de peptidases de lactocoques (daprs [53]). PepO : oligoendopeptidase ; PepI : prolyl iminopep-
tidase ; PCP : pyrrolidone carboxylyl peptidase ; PepN : PepC et PepG, aminopeptidases gnrales ;
PepL : leucyl aminopeptidase ; PepA : glutamyl aminopeptidase ; PepX : X-prolyl-dipeptidyl ami-
nopeptidase ; PepR : prolinase ; PepQ : prolidase ; PepV : dipeptidase ; PepT : tripeptidase.
Biochemical pathways during cheese ripening 309
310
Tableau III. Descripteurs et seuils de perception des acides amins [93].
bitterness; Lawrence et al. [77] have sug- flavour. Catabolism of FAA probably plays
gested that the major role of rennet in the some role in flavour development in all vari-
development of bitterness may be the pro- eties, but it is particularly significant in
duction of long peptides that will be subse- mould and smear-ripened cheeses [53]. The
quently degraded to small bitter peptides by first stage in amino-acid catabolism involves
starter proteinases. Coagulants and certain decarboxylation, deamination, transamina-
strains of starter and Penicillium spp. have tion, desulphuration or perhaps hydrolysis of
been associated with the development of the amino-acid side-chains. The second
bitterness, and it would appear that bitter- stage involves conversion of the resulting
ness in cheese results from the action of compounds (amines and -ketoacids), as
coagulant on casein with the release of bit- well as amino acids themselves, to aldehy-
ter peptides. Bitter peptides may also be pro- des, primarily by the action of deaminases
duced directly by the starter. These peptides on amines. The final stage of amino-acid
accumulate in cheese due to the absence of catabolism is the reduction of the aldehy-
proteinases or peptidases from the starter, des to alcohols, or their oxidation to acids.
or due to the inability of bitter starters to Sulphur-containing amino acids can undergo
hydrolyze them to non-bitter peptides that extensive conversion, leading to the forma-
otherwise would be too large to be perceived tion of a number of compounds, including
as bitter [79]. Low-fat cheeses have been methanethiol and other sulphur derivatives.
reported to develop bitterness [8], although General pathways for the catabolism of FAA
in full-fat cheese, a certain proportion of are summarized in Figure 10, and the subject
bitter peptides, being hydrophobic, are less has been reviewed by some researchers [6,
likely to be perceived as being bitter, per- 50, 53]. Despite the importance of these
haps due to their partition into the fat phase. pathways, detailed information on these
In addition to peptides, a number of other reactions and the agents responsible for their
compounds can contribute to bitterness in production is limited. Recently, a summary
cheese, including amino acids, amines, of the amino acid catabolic pathways iden-
amides, substituted amides, long-chain tified in LAB has been reported [24]; how-
ketones and some monoglycerides [2]. The ever, for many of these pathways, it is not
origin of unclean and related flavours in known to what extent they occur in cheese.
Cheddar has been attributed to a number of
Strecker-type compounds [34], including
phenylacetaldehyde, phenylethanol, 3-methyl- 2.1. Production of amines
butanol, 2-methylpropanol, phenol, and and pyrazines
p-cresol (see below). Off-flavours (rancid-
ity) can be due to excessive or unbalanced Decarboxylation involves the conversion
lipolysis caused by lipases/esterases from of an amino acid to the corresponding amine
starter or non-starter LAB, enzymes from with the loss of carbon dioxide. Amines,
psychrotrophs in the cheese milk, or indige- including biogenic amines, are produced in
nous milk lipoprotein lipase. cheese by enzymatic decarboxylation of
FAA [65]; the relative concentration of each
amine depends on the cheese variety and
2. CATABOLISM OF AMINO the non-starter microflora. The principal
ACIDS AND RELATED EVENTS amines in most cheeses are tyramine and
histamine (see Scheme 6) produced by
Catabolism of FAA can result in a num- decarboxylation of Tyr and His, respectively
ber of compounds, including ammonia, [53, 124], and increasing concentrations of
amines, aldehydes, phenols, indole and alco- these amines in Cheddar cheese inoculated
hols, all of which may contribute to cheese with adjunct lactobacilli has indicated that
312 P.L.H. McSweeney, M.J. Sousa
Figure 10. General pathways for the catabolism of FAA (modified from [59]).
Figure 10. Principales voies cataboliques des acides amins libres (daprs [59]).
Figure 11. The Strecker reaction between an -amino acid and an -keto acid (e.g., -ketoglutaric acid) (a), and the structure of some Strecker alde-
hydes found in cheese (b).
Figure 11. Raction de Strecker entre un acide amin et un acide ctonique (ex. acide -ctoglutarique) (a) et structure de quelques aldhydes for-
ms par la raction et retrouvs dans les fromages (b).
Biochemical pathways during cheese ripening 315
caseins than cysteine (Cys residues in the indicating that the lack of flavour in reduced-
caseins are present at low levels only in s2- fat Cheddar is due mainly to the lack of
and -caseins). Possible pathways suggested methanethiol.
in the literature for the catabolism of Met
and the structures of volatile sulphur com- Although the exact pathway for the non-
pounds found in cheese are shown in Figure enzymatic formation of CH3SH has not been
12. The catabolic products of sulphur amino established, a mechanism proposed by Man-
acids have been implicated as major con- ning [84, 85], in which a reducing agent
tributors to the flavour of Cheddar and many produces H2S from cystine/cysteine, which
other cheese varieties, but their importance then reacts with methionine to produce
in smear and surface-ripened cheeses methanethiol, is accepted by many investi-
appears to be accentuated by their high con- gators [2, 53, 59].
centration in the surface smear [2, 14, 34, The concentration of H 2 S in cheese
57]. Low molecular weight sulphur com- increased during ripening, and initially no
pounds in cheese include methanethiol correlation was found between its concen-
(CH 3 SH), hydrogen sulphide (H 2 S), tration (or those of other sulphur com-
dimethylsulphide (DMS; CH 3 SCH 3 ), pounds) and flavour development [6],
dimethyldisulphide (DMDS; CH3SSCH3), although Barlow et al. [11] found a high
dimethyltrisulphide (DMTS; CH3SSSCH3) correlation between the concentration of
and carbonyl sulphide (O = C = S). Sulphur H2S and flavour, particularly when the value
compounds are thought to interact with each for H2S was combined with either the con-
other and with other compounds in cheese, centration of water-soluble nitrogen (WSN)
generating other volatile flavour compounds or lactic acid. These authors suggested that
[69]. these parameters (H2S and WSN/lactate)
Methanethiol is present in Camembert were better predictors of the future quality of
together with other sulphur compounds, such young cheeses than their flavour or compo-
as 2,4-dithiapentane, 3,4-dithiahexane, sition at an early age. The presence of H2S
2,4,5-trithiahexane and 3-methylthio-2,4- in fresh curd indicates that some may also be
dithiapentane, and these compounds are formed during pasteurization of cheese milk
responsible for the garlic note which can be [74]. DMS, DMDS and DMTS are thought
found in well-ripened Camembert cheese to be important contributors to cheese
[3]. Br. linens is one of the principal flavour [10, 14]. DMS is a product of the
microorganisms found on the surface of metabolism of propionic acid bacteria
smear-ripened cheeses, is also present on formed from Met, and is a component of
the surface of mould-ripened varieties (e.g., Swiss cheese flavour [2]. DMS concentra-
Camembert), and can produce methanethiol tions in Cheddar cheese remain constant for
enzymatically. In cheeses such as Cheddar, up to 6 months, but decrease thereafter [6].
which lacks a surface microflora, flavour is DMDS can be formed as an end-product of
produced by starter and non-starter bacte- Strecker degradation, and has been identified
ria and their enzymes, and the production in Parmesan [10], Cheddar [6, 11], and sur-
of methanethiol is thought to be a chemical face-ripened cheeses [64]. DMDS concen-
process [69]; however, Urbach [139] sug- trations in Cheddar correlate reasonably well
gested that the secondary flora, particularly with flavour scores [11]. DMTS has been
in Cheddar and Emmental, are likely to be associated with the aroma of cooked cab-
more important than chemical reactions for bage, broccoli, or cauliflower, and has been
the formation of sulphur compounds. Dimos identified in Parmesan [10] and Cheddar
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methanethiol in full-fat and reduced-fat produced directly from methanethiol, but it
Cheddar was highly correlated with flavour, is unclear how DMTS is produced in cheese.
316
P.L.H. McSweeney, M.J. Sousa
Figure 12. Sulphur
compounds which
have been identified
in cheese and possi-
ble pathways sug-
gested in the litera-
ture for the formation
of some compounds
from methionine.
Figure 12. Composs
sulfurs identifis
dans les fromages et
possibles voies de
transformation de la
mthio-nine.
Biochemical pathways during cheese ripening 317
Figure 13. Pathways for the catabolism of tryptophan (a), phenylalanine (b) and tyrosine (c), by
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arrows: non-enzymatic chemical reactions; dashed arrows: unknown mechanism; and Atase: amino-
transferase.
Figure 13. Catabolisme du tryptophane (a), de la phnylalanine (b) et de la tyrosine (c), par les
levains acidifiants, les levains additionnels et les bactries non-levains (daprs [24]). Flches en
gras : raction enzymatiques ; flches non en gras : ractions chimiques non enzymatiques ; flches
en pointills : mcanisme inconnu ; Atase : aminotransfrase.
Biochemical pathways during cheese ripening 319
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