ARTICLE IN PRESS

International Dairy Journal 16 (2006) 555–562

www.elsevier.com/locate/idairyj

Review

Lipoprotein lipase and lipolysis in milk

Hilton C. Deeth
School of Land and Food Sciences, The University of Queensland, Brisbane 4072, Australia
Received 19 April 2005; accepted 26 August 2005

Abstract

Bovine milk contains a lipoprotein lipase that accounts for most, if not all, of its lipolytic activity. The total lipase activity in raw milk
is sufficient to cause rapid hydrolysis of a large proportion of the fat. However, in reality this does not happen, because the lipase is
prevented from accessing the fat by the milkfat globule membrane. Physical damage to this membrane in raw milk initiates lipolysis.
Furthermore, simply cooling certain individual milks soon after secretion can initiate the so-called spontaneous lipolysis. The
biochemical basis of spontaneous lipolysis is still poorly understood, but it appears to be related to a balance between activating and
inhibiting factors in the milk. Lipolysis in milk and milk products causes rancid off-flavours and other problems, and is a constant
concern in the dairy industry. A thorough understanding of the mechanism of lipolysis and constant vigilance by operatives is required to
minimize lipase-related problems.
r 2006 Elsevier Ltd. All rights reserved.

Keywords: Lipoprotein lipase; Milk lipase; Lipolysis; Free fatty acids

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 555
2. Milk lipase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 556
3. Lipolysis in milk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 557
3.1. Spontaneous lipolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 557
3.1.1. Mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 557
3.1.2. Practical aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 558
3.2. Induced lipolysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 558
3.2.1. Mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 558
3.2.2. Practical aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 559
4. Comparison of lipoprotein lipase and bacterial lipase . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 559
5. Analytical methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 560
6. Challenges. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 560
7. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 561
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 561
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 561

1. Introduction

Lipases are enzymes that catalyse the hydrolysis of

Tel.: +61 7 3346 9191; fax: +61 7 3365 1177. triglycerides (triacylglycerols), the major lipid components
E-mail address: h.deeth@uq.edu.au. of milk. This hydrolysis is commonly referred to as

0958-6946/$ - see front matter r 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.idairyj.2005.08.011
 ARTICLE IN PRESS
556 H.C. Deeth / International Dairy Journal 16 (2006) 555–562
lipolysis. The products of the reaction are free, or non-
esterified fatty acids, and partial glycerides (mono- and
diglycerides) and, in some cases, glycerol. By definition,
lipases act at the lipid–water interface of emulsions of long-
chain, insoluble triglycerides, while the related esterases act
on esters of short-chain fatty acids (and other acids) and
soluble esters (Jaeger et al., 1994; Okuda & Fujii, 1968).
The significance of lipolysis in milk is two-fold: flavour
production and altered functionality. Free fatty acids
(FFAs), particularly those of short and medium chain
length, have strong flavours that, in most cases, are
considered undesirable. The flavour of whole milk with
an elevated FFA level, 4
1.5 mmol L1, is unacceptable
to most people (IDF, 1987) and is variously described as
rancid, butyric, astringent or even bitter, although this last
flavour is usually attributable to products of proteolysis
not lipolysis. The term ‘‘rancid’’ is also used to describe the
off-flavour due to lipid oxidation, but the two types of
rancidity are distinctly different in origin and flavour. In
general, oxidative and hydrolytic rancidities are not
related, although there is some evidence that FFAs are
more susceptible to oxidation than the parent triglycerides
(Aubourg, 2001). Flavours due to lipolysed fat can also be
desirable; some cheeses such as certain hard Italian and
Blue varieties owe their characteristic flavour to the
presence of FFAs (Deeth & Fitz-Gerald, 1995).
A major functionality effect of lipolysis in milk is
depression of its foaming ability when injected with steam.
Fig. 1 demonstrates the relationship between foaming
capacity (steam frothing value) and degree of lipolysis (as
120
100
80
Steam Frothing Value
60
40
20
0 hydrolysis of fatty acids from the sn-1 and sn-3 positions
0 0.5 1 1.5 2 2.5 3 3.5
FFA (mmol L-1)
Fig. 1. Relationship between the steam frothing value (SFV) and free fatty
acid (FFA) level of milks. SFV ¼ volume of froth  100/original volume
of milk (redrawn from Deeth and Smith (1983) with permission).
FFA) (Deeth & Smith, 1983). This effect is manifested in
difficulty in producing an acceptable foam when making
cappuccino coffee. It is due to the partial glycerides
produced during lipolysis, which are surface active and
displace the foam-stabilizing proteins at the air–water
interface of the foam bubbles (Buchanan, 1965). Some
other functionality defects, such as impaired creaming
ability during separation and increased churning time in
the manufacture of butter, have been reported (Deeth &
Fitz-Gerald, 1995).
2. Milk lipase
Despite early research suggesting otherwise (Downey &
Andrews, 1969), it is now accepted that only one lipolytic
enzyme, lipoprotein lipase (LPL), accounts for most, if not
all, of the lipolytic activity in bovine milk (Olivecrona,
1980; Olivecrona, Vilaro, & Olivecrona, 2003). This is not
the case for human milk, which contains a bile-salt-
stimulated lipase in addition to LPL. LPL is involved in
the synthesis of milkfat triglycerides in the mammary gland
and its presence in milk is due to a spillover from this
gland. Bovine milk LPL does not serve any known
biological purpose in milk. By contrast, the bile-salt-
stimulated lipase in human milk appears to have a major
role in digestion of milkfat by the newborn (Fredrikzon,
Hernell, Bläckberg, & Olivecrona, 1978).
LPL is a glycoprotein with two N-linked oligosacchar-
ides, which appear to be necessary for the activity of the
enzyme (Olivecrona et al., 2003). It exists as a homodimer
with a molecular mass of
100 kDa (Kinnunen, Huttunen,
& Ehnholm, 1976). In milk, it is normally associated with
the casein micelle. This association is largely electrostatic as
the enzyme can be released from the micelle by sodium
chloride or heparin. The negatively charged heparin has a
high affinity for LPL (Hoynes & Downey, 1973; Olivecrona
et al., 2003); heparin–sepharose affinity chromatography is
used for the separation and purification of the enzyme from
milk (Kinnunen et al., 1976). Hydrophobic interactions
between LPL and casein are also important, as the enzyme
can be released with dimethylformamide (Fox, Yaguchi, &
Tarassuk, 1967).
LPL is relatively unstable to heat. High-temperature,
short-time (HTST) pasteurization (72 1C for 15 s) inacti-
vates most, if not all, of the enzyme in milk (Farkye,
Imafidon, & Fox, 1995), and therefore LPL causes little, if
any, lipolysis in pasteurized milk and products derived
from pasteurized milk. It is also unstable to acid and hence
would normally be inactivated in the stomach.
LPL exhibits positional specificity but not fatty acid
specificity (Morley & Kuksis, 1977); it catalyses the
of the triglyceride molecule (Somerharju, Kuusi, Paltauf, &
Kinnunen, 1978). Fatty acids are only released from the
2-monoglyceride after rearrangement to a 1(3)-monogly-
ceride (Nilsson-Ehle, Egelrud, Belfrage, Olivecrona, &
Borgström, 1973). Because the short-chain fatty acids are
 ARTICLE IN PRESS
H.C. Deeth / International Dairy Journal 16 (2006) 555–562
concentrated in the sn-3 position of bovine milk triglycer-
ides, the profile of FFAs released by LPL may give the
impression of preferential release of short-chain fatty acids
(Ouattara, Jeon, Hart-Thakur, & Schmidt, 2004).
The optimum conditions for measuring milk LPL
activity using an emulsified triglycerides substrate contain-
ing long-chain fatty acids (e.g., ultra-high-temperature
(UHT)-treated (homogenized) cream) are pH
9.0, tem-
perature
37 1C, with a blood serum lipoprotein activator
(e.g., bovine or human blood serum) and a Ca2+ salt as a
fatty acid acceptor to minimize product inhibition. The
total LPL activity in raw milk measured under these
conditions suggests that there is sufficient activity to
hydrolyse the fat in milk in a relatively short time. It has
been reported that the typical LPL activity in milk,
measured in an assay at pH 7, is
100 nmol min1 mL1
(Olivecrona et al., 2003); if lipolysis in milk proceeded at
this rate, the milk would be unacceptably rancid in less
than 10 min. Lipolysis does not proceed rapidly because the
milkfat is in the form of globules 0.2–10 mm in diameter,
which are enveloped in the protective milkfat globule
membrane (MFGM). In freshly secreted milk, this
biological membrane is intact and forms an effective
barrier to access by milk lipase to the fat. However, in
certain situations, this protection is reduced or completely
eliminated, and lipolysis proceeds.
3. Lipolysis in milk
Lipolysis in milk can be broadly categorized into
two types: spontaneous and induced. Spontaneous lipo-
lysis is initiated by the simple act of cooling raw milk to
o10 1C soon after it is taken from the cow. By contrast,
induced lipolysis is initiated by physical damage to the
MFGM, which allows the lipase access to the fat sub-
strate. Physical actions causing induced lipolysis include
agitation, pumping and homogenization. Both sponta-
neous and induced lipolysis progress during sub-
sequent storage of the milk, with most occurring in the
first 24 h when the milk is refrigerated (Ouattara et al.,
2004).
3.1. Spontaneous lipolysis
3.1.1. Mechanism
Three biochemical factors appear largely to determine
the susceptibility of milk to spontaneous lipolysis: the
amount of lipase activity; the integrity of the MFGM; and
the balance of lipolysis-activating and -inhibiting factors
(Cartier & Chilliard, 1990; Deeth & Fitz-Gerald, 1975;
Sundheim, 1988). These have been demonstrated in
experiments where two of these variables have been kept
constant and the other varied. Intuitively, one would
expect the amount of lipase activity in milk to be a major
factor; however, it has to be considered that raw milk has
more than enough lipase activity to cause extensive
lipolysis under favourable conditions. Furthermore, a poor
557
correlation has been found between lipase activity and the
level of spontaneous lipolysis in milks (Cartier & Chilliard,
1990). Similarly, the integrity or fragility of the MFGM
would appear to be an obvious factor in predisposing a
milk to spontaneous lipolysis. While it can be shown to be
a factor, it cannot explain the major difference between
milk susceptible to spontaneous lipolysis and ‘‘normal’’
milks (Cartier & Chilliard, 1990; Sundheim & Bengtsson-
Olivecrona, 1987).
Components in the skim milk fraction other than LPL
have been shown to be important in spontaneous lipolysis.
It has been known for some time that addition of normal
milk to milk susceptible to spontaneous lipolysis, soon
after secretion from the cow, has an inhibitory effect on
subsequent spontaneous lipolysis. That is, the level of
FFAs produced is less than the average of the levels
produced by both milks separately (Dunkley & Smith,
1951). This effect has been attributed to the presence of an
inhibiting factor or factors in normal milk and a deficiency
of such factors in susceptible milk. However, it was also
discovered that mixing two milks, which were susceptible
to spontaneous lipolysis, but to different degrees, could
result in more lipolysis than the average of the two separate
milks. This suggested the presence of activating factors in
the most susceptible milk. It therefore appears that milk
contains both activatory and inhibitory components, and
that the activator–inhibitor balance largely determines the
susceptibility of a milk to spontaneous lipolysis (Deeth &
Fitz-Gerald, 1975).
The most likely candidates for activatory factors are
(apo)lipoproteins. Their presence in milk has been demon-
strated (Castberg & Solberg, 1974; Anderson, 1979), and it
is known that addition of these blood components or whole
blood serum (Jellema, 1975) to milk initiates lipolysis.
However, such lipolysis proceeds without the need to cool
the milk, a prerequisite of spontaneous lipolysis. Heparin-
like glycosaminoglycans in milk may also be activatory
(Iverius & Östlund-Lindqvist, 1976). Inhibitory factors
have also been discovered in milk; proteose peptone
fraction PP3 inhibits lipolysis (Anderson, 1981; Cartier,
Chilliard, & Paquet, 1990; Girardet, Linden, Loye,
Courthaudon, & Lorient, 1993), as does a peptide isolated
from the MFGM, which is very similar to PP3 (Shimizu,
Miyaji, & Yamauchi, 1982).
The susceptibility of milk to spontaneous lipolysis,
measured by the level of FFAs developed in the milk
after overnight storage at low temperature, shows a
high correlation with the amount of lipase activity
which becomes associated with the milkfat globule
(Ahrné & Björck, 1985; Sundheim & Bengtsson-
Olivecrona, 1985). In highly susceptible milks, a large
proportion of the lipase moves from the skim phase to the
cream phase. The extent of this redistribution appears to be
largely governed by the activator–inhibitor balance—
activators promote the attachment of the lipase to the
MFGM, while inhibitors prevent the attachment. Cooling
the milk soon after it leaves the cow triggers this
 ARTICLE IN PRESS
558 H.C. Deeth / International Dairy Journal 16 (2006) 555–562
attachment as it does not occur in uncooled susceptible
milk, nor does such milk undergo lipolysis. The exact
nature of the attachment and the manner in which the
activatory and inhibitory factors influence the attachment
are presently unknown.
3.1.2. Practical aspects
Spontaneous lipolysis occurs at the farm only. The
milk of some individual cows is particularly susceptible
to this type of lipolysis and its FFA level may reach as
high as 10 mmol L1 after refrigerated storage for
24 h
(Deeth & Fitz-Gerald, 1995). On the other hand, the milk
of other cows is resistant to spontaneous lipolysis
and the FFA level may remain p0.5 mmol L1 during
storage (Bråthen, 1980; Connolly, Murphy, O’Connor, &
Headon, 1979); milk that remains at a low FFA level
is sometimes referred to as ‘normal’ milk. Milks of
other cows may develop FFA levels between these two
extremes. Individual cow milks vary in their susceptibi-
lity to spontaneous lipolysis from day to day, although,
in general, cows that produce ‘normal’ milk do so
consistently.
The major factors associated with spontaneous lipolysis
are late lactation (Chazal & Chilliard, 1986), poor-quality
feed (Jellema, 1980) and mastitis (Downey, 1980). Sponta-
neous lipolysis becomes most noticeable in the milk from
farms with a majority of cows in late lactation at a time
when the cows’ plane of nutrition is low. Therefore,
districts where seasonal calving is practised may encounter
this problem during periods of poor feed availability.
Lipolysis in late lactation milk may be initiated by
blood serum components, such as activatory lipoproteins,
leaking into the milk through leaky tight junctions between
lactating cells (Anderson, 1979; Castberg & Solberg,
1974).
Mastitis can contribute to spontaneous lipolysis. How-
ever, in this case, it is necessary to distinguish between an
elevated initial FFA level (i.e., at secretion) and the FFAs
developed during storage of the milk. The former occurs
because the infection and associated inflammation disrupts
milk synthesis, and a higher level of fatty acids remain
unesterified than in healthy glands. However, it has been
shown that spontaneous lipolysis also occurs in mastitic
milk and contributes to the final FFA level (Fitz-Gerald,
Deeth, & Kitchen, 1981).
A variation of the classic spontaneous lipolysis is
temperature-induced lipolysis. This occurs when precooled
milk is warmed to 25–35 1C and then recooled to o10 1C
(Claypool, 1965; Kon & Saito, 1997). Following this
‘‘temperature activation’’, lipolysis proceeds in much the
same way as spontaneous lipolysis. In practice, this can
occur when warm milk straight from the cow is mixed with
precooled milk, for example, milk in a refrigerated bulk
farm vat from a previous milking. It has been shown that
milk that is susceptible to spontaneous lipolysis is more
susceptible to temperature-induced lipolysis than normal
milk (Saito & Kim, 1995).
3.2. Induced lipolysis
3.2.1. Mechanism
The mechanism of induced lipolysis is more easily
explained in terms of the known action of lipase than that
of spontaneous lipolysis. In contrast to other enzymes, the
rate at which a lipase-catalysed reaction proceeds is not
governed by the substrate concentration at the interface
between the lipid substrate and the aqueous phase of an
emulsion (Jaeger et al., 1994). In induced lipolysis in milk,
the physical forces involved disrupt the milkfat globule and
expose the lipid substrate to the lipase; the more severe the
treatment or the longer time it is applied, the greater the
disruption and resulting surface area of the exposed or
‘‘free’’ fat.
The concept of ‘‘free fat’’ is somewhat controversial as it
can have different meanings according to how it is
measured (Evers, 2004). Two methods that have been used
are warming the milk and centrifuging to reveal a layer of
molten fat on the top of the milk, and extraction with an
organic solvent, which is unable to extract fat from natural,
intact fat globules. Both methods have been criticized, a
major concern being the ability of the procedures
themselves to disrupt fat globules and create free fat
(Evers, 2004; Evers, Crawford, Robinson, Harding, &
McCarthy, 2001). However, some workers have success-
fully used a non-polar solvent such as hexane to extract
free fat (Deeth & Fitz-Gerald, 1978). An alternative
method uses an exogenous lipase, which is unable to
attack intact fat globules; the amount of lipolysis produced
under specified conditions gives a measure of lipase-
accessible fat (Cartier & Chilliard, 1990; Deeth & Fitz-
Gerald, 1978).
Agitation of milk can have two effects on the fat in milk
and these have implications for the amount of lipolysis
induced. At relatively low-intensity treatments, such as
low-speed agitation, the fat globules coalesce (or churn as
occurs in butter making), while under high-intensity
conditions, the fat globules are dispersed and form much
smaller globules, in a manner similar to homogenization
(Deeth & Fitz-Gerald, 1977). While the extent of fat
globule membrane damage may be similar in both cases,
the extent of lipolysis resulting from the latter type is much
greater because the surface area of the lipase-accessible fat
is much greater. Conversely, the amount of free fat,
measured by either warming and centrifuging or solvent
extraction, is greater in the milk subjected to the former,
low-intensity agitation. Therefore, the level of free fat
measured by these two common methods may not be a
good indicator of the susceptibility of a milk to induced
lipolysis.
The temperature of agitation also influences its effect on
the fat globule, with fat globule coalescence being favoured
at low temperatures and homogenization at high tempera-
tures. During low-intensity agitation at low temperature, a
considerable proportion of the lipase in milk becomes
associated with the cream fraction, presumably attached to
 ARTICLE IN PRESS
H.C. Deeth / International Dairy Journal 16 (2006) 555–562 559

the MFGM (Deeth & Fitz-Gerald, 1977). In contrast to the 5

situation in spontaneous lipolysis, the extent of lipolysis
induced under these conditions does not correlate well with
the amount of attached lipase.
4.5

3.2.2. Practical aspects

In the dairy industry, induced lipolysis can occur on the
4
farm, during transport or in the factory (Deeth & Fitz-
Gerald, 1995). The major causes are:

 agitation and pumping, especially with air incorporation 3.5

or high shear;

Acid Degree Value (mmol 100 g fat-1)

 homogenization;
 mixing raw and homogenized milk; and 3
 freezing/thawing of milk.

Agitation of raw milk in such a way that the milkfat

2.5
globule is damaged is an effective means of inducing
lipolysis; in the laboratory, this can be accomplished using
a blender. Incorporation of air, with resulting froth
formation, is an important factor in this type of lipolysis 2
activation. The amount of lipolysis initiated in this way is
very temperature dependent, as illustrated in Fig. 2, which
shows two distinct maxima separated by a minimum (Fitz-
1.5
Gerald, 1974). The shape of this curve changes with the
intensity of the agitation treatment.
On farms, a significant cause of induced lipolysis is
excessive air intake at the teat cups, which causes milk to 1
surge in the milk lines, particularly in the ‘‘risers’’ of high-
line installations (Evers & Palfreyman, 2001; O’Brien,
O’Callaghan, & Dillon, 1998). In factories, excessive
0.5
pumping and use of centrifugal pumps sucking air through
faulty seals are the major causes (Downes, Nieuwoudt, &
Slabbert, 1974).
Homogenization is a very effective way of inducing 0
lipolysis in raw milk (Mulder & Walstra, 1974). In splitting 0 10 20 30 40 50 60
the natural fat globules into much smaller fat particles and Temperature (°C)
coating them with milk proteins, rather than the natural Fig. 2. Relationship between degree of lipolysis (acid degree value) and
MFGM, the LPL is brought into intimate contact with fat temperature of activation in precooled whole raw milk (redrawn from
with a large surface area. However, in practice in the dairy Fitz-Gerald (1974) with permission).
industry, milk is homogenized either immediately before or
after pasteurization. Since pasteurization inactivates the (Datta, Hayes, Kelly, & Deeth, 2005); the mechanism of
lipase, lipolysis does not occur in (pasteurized) homo- this activation has not been elucidated.
genized milk. Freezing and thawing disrupts the MFGM and allows
The effect of homogenization on natural milkfat access to the fat by LPL (Willart & Sjöström, 1966). The
globules as a substrate for LPL can be demonstrated by effect of this treatment has been demonstrated by several
mixing (pasteurized) homogenized milk and raw milk researchers who have used it to isolate the membrane from
(Nilsson & Willart, 1960). This initiates lipolysis and can (washed) cream (Walstra, 1985).
render the mixture rancid within 1–2 h at room tempera-
ture. Industry problems can arise from this phenomenon
when well-meaning operatives with no knowledge of the 4. Comparison of lipoprotein lipase and bacterial lipase
LPL system return excess homogenized milk to a bulk raw
milk supply at the end of a day’s operation. In the dairy industry, not all undesirable lipolysis is
In addition to affecting the milkfat globules in milk, caused by LPL. During cold storage of milk, psychro-
high-pressure homogenization also appears to activate the trophic bacteria such as Pseudomonas grow and produce
enzyme, as has been recently demonstrated in bovine milk enzymes such as lipases and proteases, which have major
 ARTICLE IN PRESS
560 H.C. Deeth / International Dairy Journal 16 (2006) 555–562
Table 1
Comparison of the characteristics of milk lipoprotein lipase (LPL) and lipases from psychrotrophic bacteria
Milk LPL
Destroyed by high temperature-short time (HTST) pasteurization
Milk fat globule membrane (MFGM) acts a barrier to lipid
substrate
Activated by serum lipoproteins
Effect mostly associated with fresh milk and cream
Effect in cheese/butter obvious at manufacture and does not
change during storage
High levels in (raw) milk
effects on the quality of milk and milk products (Sørhaug
& Stepaniak, 1997). The lipases produced by these bacteria
have different characteristics from LPL and it is important
to recognize these differences in order to be able to
determine the cause of lipolysis in particular situations.
A comparison of the two types of lipase is given in Table 1.
A major difference in their mode of action is that the
MFGM does not appear to be a barrier to access of the
bacterial lipases to the fat in intact fat globules (Fitz-
Gerald & Deeth, 1983). While this is true for the lipases
produced by psychrotrophic bacteria, some other microbial
lipases do not have this characteristic. For example, the
lipase produced by Candida cylindracea cannot access the
fat in intact fat globules, a property exploited in the test for
free or lipase-accessible fat (Cartier & Chilliard, 1990;
Deeth & Fitz-Gerald, 1978). The reason for this difference
between lipases is unclear.
Another significant difference is their heat stabilities.
Normal HTST pasteurization inactivates LPL, while most
of the lipases from psychrotrophic bacteria are heat
resistant (Fitz-Gerald & Deeth, 1983), with many of them
surviving even intense heat treatments such as UHT
treatments (
140 1C for 4 s) (Christen, Wang, & Ren,
1986). Because only trace amounts of these heat-resistant
enzymes are likely to be produced in milk during normal
commercial operations, their effects are only observed in
products such as UHT milk, butter, cheese and milk
powders after a period of storage. Affected products, or
products manufactured from them, develop rancid off-
flavours and may become unacceptable for human
consumption.
5. Analytical methods
The major analysis performed in relation to LPL and
lipolysis is for total FFA. This is usually performed by
either titrating aliquots of demulsified fat from milk or by
titrating organic solvent extracts of milk using an alcoholic
alkali. The classic method of the former type is the BDI
method (Evers, 2003), which gives the FFA level as the acid
degree value in mmol 100 g fat1, whereas the results from
the solvent extraction methods are expressed in mmol L1
or meq L1. Individual fatty acids can be determined by
Lipases from psychrotrophic bacteria
Stable to HTST and even to ultra-high temperature (UHT) treatment
MFGM presents no barrier
Most not activated by serum lipoproteins
Effect mostly associated with stored products—UHT milk, cheese, butter, milk
powders
Effect in cheese/butter obvious only after storage
Trace levels only
gas chromatography and a measure of total FFA can be
obtained by summing the concentrations of the individual
FFA (e.g., De Jong & Badings, 1990). The methods to
measure FFA have been reviewed by several authors (e.g.,
Collomb & Spahni, 1995; Deeth & Fitz-Gerald, 1995; IDF,
1991; Kuzdzal-Savoie, 1980).
The other relevant analytical method is for lipase
activity. In contrast to FFA analyses, this has little use in
quality control programs because the level of milk lipase
activity in raw milk is not directly related to any quality
issue and the enzyme is inactivated by pasteurization.
However, there is considerable interest in assaying for
bacterial lipases. Even trace levels of these enzymes can
cause severe problems in stored products, and hence their
analysis presents a major challenge. Several methods have
been proposed, but to date no one method has been shown
to be ideal or widely adopted (Deeth & Touch, 2000).
6. Challenges
Challenges in relation to LPL and lipolysis can be
considered in terms of the dairy industry or dairy scientists.
For the dairy industry, continuing education of the work
force is the major challenge. Knowledge of the conditions
under which lipolysis, either spontaneous or induced,
caused by LPL can be initiated is necessary if problems
are to be avoided. The severe consequences of seemingly
innocuous actions such as mixing homogenized milk and
raw milk illustrate this need. The other challenge for the
industry is to recognize whether a problem with lipolysis is
the result of the action of LPL or bacterial lipase, as each
of these requires quite different remedial action.
Several challenges still await the dairy scientist. These
include elucidation of the activators and inhibitors in milk,
which determine the extent of spontaneous lipolysis. An
important aspect of this is the physiological determinants
of the balance and imbalance of these in the milk from
certain cows under certain conditions. The most relevant
situations are late lactation and poor-quality feed intake.
A related issue in spontaneous lipolysis, which has not been
adequately explained, is the nature of the association of the
lipase with the MFGM and the form of the enzyme when
attached. Another challenge that has arisen is to determine
 ARTICLE IN PRESS
H.C. Deeth / International Dairy Journal 16 (2006) 555–562
the mechanism for the apparent increase in activity of LPL
when subjected to high-pressure homogenization.
7. Conclusion
Lipoprotein lipase is an important indigenous enzyme in
bovine milk. While it does not appear to have a
physiological function for the calf, it can cause major
quality problems in the dairy industry. Production of
rancid off-flavours and depression of the steam frothing
capacity of milk are among these problems. A large body
of knowledge of the enzyme and the circumstances under
which its action in milk is initiated has accumulated over
many years. However, there remain on-going challenges for
the dairy industry in controlling the problem and for the
dairy scientist in elucidating the remaining details of the
mechanisms by which lipolysis is promoted and proceeds.
Acknowledgements
The author gratefully acknowledges the assistance of
Dr. Nivedita Datta and Ms. Carolyn Fitz-Gerald in the
preparation of this paper.
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