MEAT

SCIENCE
Meat Science 77 (2007) 7–16
www.elsevier.com/locate/meatsci

Mechanisms controlling pork quality development: The

biochemistry controlling postmortem energy metabolism
T.L. Scheffler, D.E. Gerrard *

Department of Animal Sciences, Purdue University, 915 W. State Street, West Lafayette, IN 47907, United States

Received 23 March 2007; received in revised form 2 April 2007; accepted 2 April 2007

Abstract

Pale, soft and exudative (PSE) pork represents considerable economic losses for the industry due to its limited functionality and unde-
sirable appearance. During the past several decades, exhaustive research covering various aspects of the food chain has established geno-
typing procedures, recommended handling practices, and quality indicators in order to reduce the incidence of inferior pork quality.
Despite these efforts, there is still a relatively high occurrence of PSE pork. Development of pork quality attributes is largely governed
by the rate and extent of postmortem pH decline. The combination of high temperature at low pH or abnormally low ultimate pH causes
denaturation of sarcoplasmic and myofibrillar proteins, resulting in paler color and reduced water holding capacity. The pH decline is
closely related to muscle energy availability and demand at or around slaughter. The postmortem degradation of glycogen through gly-
cogenolysis and glycolysis provides ATP to help meet energy demand and decreases pH by generating lactate and H+. Therefore, the flux
of metabolites through glycolysis, the involvement of energy signaling pathways that modulate glycolytic activity, and the inherent
metabolism of different fiber types are critical factors influencing pH decline and pork quality. Further, recent work implicates adenosine
monophosphate-activated protein kinase (AMPK) as a major energy sensor for the cell, and thus may be involved in the control of post-
mortem metabolism. The intent of this paper is to review the biochemistry controlling postmortem energy metabolism in pig muscle and
explore new information generated using genetic mutations in order to define the fundamental mechanisms controlling the transforma-
tion of muscle to meat.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Pork; Biochemistry; Glycolysis

1. Introduction texture due to enhanced water binding capability. Con-

Increased health-consciousness among consumers has
led to significant changes in pork carcass attributes over
the past several decades. Swine producers have focused
selection pressure on improving carcass merit, resulting in
increased lean growth and higher yielding carcasses. How-
ever, extreme variation in pork quality, ranging from dark,
firm, and dry (DFD) to pale, soft and exudative (PSE)
pork, has also become more prevalent. DFD pork has a
dark, unattractive appearance and a firm, dry, and sticky
*
Corresponding author. Tel.: +1 765 494 8280; fax: +1 765 494 6816.
E-mail address: dgerrard@purdue.edu (D.E. Gerrard).
versely, PSE pork is characterized by pale color, soft tex-
ture, and low water-holding capacity, and has limited
functionality in further processing. Surveys indicated that
in pork produced in the USA in 2003, 15.5% was PSE
(Stetzer & McKeith, 2003). The inferior quality attributes
of PSE pork are estimated to cost the pork industry $100
million annually (Carr, Kauffman, Meeker, & Meisinger,
1997).
Due to the considerable economic losses, PSE pork has
been a significant source of research attention. Early stud-
ies established that the development of PSE pork was lar-
gely due to an increased rate of early postmortem
glycolysis, indicated by elevated muscle temperature and
rapid pH decline (Briskey, 1964). More recently, the extent

0309-1740/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2007.04.024
8 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
of glycolysis and ultimate pH have been implicated (Sellier
& Monin, 1994). Further research has established measur-
able indicators of quality, and yielded molecular diagnostic
techniques and control methods. Yet, despite the extensive
efforts to improve pork quality, Cassens (2000) concluded
that little progress has been made. A greater understanding
of postmortem metabolism during the conversion of mus-
cle to meat and the mechanisms of enhanced and extended
glycolysis are critical to characterizing pork quality
development.
2. Conversion of muscle to meat
Transitions between rest and exercise require that mus-
cle is a dynamic tissue with the ability to adapt to dramatic
changes in energy expenditure. Muscle contraction is a
rapid, energetically demanding process that requires the
splitting of adenosine triphosphate (ATP) in order to meet
energy requirements for muscle contraction and relaxation,
sequestration of calcium and maintenance of ion gradients.
The most efficient means of generating ATP is through
mitochondrial oxidative metabolism, but the ATP concen-
tration in muscle supplies enough energy for only a few
twitches. Therefore, additional reactions must be able to
buffer energy levels when other metabolic processes are
not able to meet ATP demand.
Phosphocreatine (PCr) is present at a higher concentra-
tion (18–19 lmole/g) than ATP (6.6–6.8 lmole/g) in pig
longissimus (Bendall, 1973) and can be quickly utilized in
a chemical reaction to rephosphorylate ADP to ATP by
the enzyme creatine kinase. Additionally, myokinase cata-
lyzes the conversion of two adenosine diphosphate (ADP)
to adenosine monophosphate (AMP) and ATP. Together,
these reactions allow muscle to quickly increase energy pro-
duction in order to keep cellular ATP constant and main-
tain homeostasis. Aerobic metabolism and PCr are able
to satisfy energy demand when oxygen is adequate and
muscle is working slowly, but if contraction proceeds rap-
idly, oxygen becomes limiting and the muscle will resort to
anaerobic glycolysis to supply the energy needed for con-
traction. Similarly, the removal of blood during the slaugh-
tering process eliminates the oxygen supply and forces
metabolically active muscles to adapt to new physiological
circumstances.
The basic biochemical reactions and physical changes
underlying the conversion of muscle to meat are well recog-
nized. ATP production is necessary to keep the muscle in
the relaxed state, but postmortem muscle has a high rate
of ATP turnover (Bate-Smith & Bendall, 1949). Initially,
ATP is replenished by the creatine kinase and myokinase-
catalyzed reactions. Once 70% of the PCr pool has been
degraded, ATP levels rapidly decline (Bendall, 1951), and
muscle glycogen must be degraded (glycogenolysis) and
metabolized in anaerobic glycolysis in order to rephospho-
rylate ADP to ATP and prevent the formation of perma-
nent actomyosin crossbridges. Anaerobic glycolysis also
produces lactate, H+, and heat. Because postmortem mus-
cle does not have the means to remove waste products, lac-
tate and H+ accumulate and lower the pH. As the
breakdown of ATP exceeds its synthesis by glycolysis, less
ATP is available and the formation of actomyosin bonds
shortens sarcomeres and increases muscle tension, signal-
ing the onset of rigor mortis. Rigor mortis is complete
when the ATP supply is exhausted; thus, actomyosin cross-
bridges cannot be broken and the muscle is relatively inex-
tensible. Muscle tension will eventually decrease with
postmortem storage as a result of degradation of myofibr-
illar proteins and loss of structural integrity.
The rate and extent of pH decline during the conversion
of muscle to meat significantly impact the development of
fresh meat quality attributes (Fig. 1). Normally, pH
declines gradually from 7.4 in living muscle to roughly
5.6–5.7 within 6–8 h of postmortem and then has an ulti-
mate pH at 24 h (pHu) of about 5.3–5.7 (Briskey & Wis-
mer-Pedersen, 1961). However, muscles with a hastened
pH decline exhibit rapid glycolysis and produce large
amounts of heat, which slows carcass chilling. This results
in a rapid muscle pH decline to less than 6.0 during the first
hour after slaughter and an ultimate pH of 5.3–5.7. The
onset of rigor mortis at high temperature and low pH
causes the denaturation of approximately 20% of the sarco-
plasmic and myofibrillar proteins (Honikel & Kim, 1986),
and the reduction in myosin head length is sufficient to
draw thick and thin filaments closer together, leading to
increased expulsion of water (Offer et al., 1989). Greater
precipitation of sarcoplasmic proteins is largely responsible
for paler pork color, while denaturation of myofibrillar
proteins explains the reduced water holding capacity in
PSE muscle (Joo, Kauffman, Kim, & Park, 1999). In con-
trast, an extended pH decline proceeds at a normal rate
but continues to a low pHu of roughly 5.3–5.5, resulting
in ‘‘acid meat’’. Abnormally low pH reduces the net charge
of myofibrillar proteins, and the attraction moves filaments
closer together and forces water out of the myofilament lat-
tice (Irving, Swatland, & Millman, 1989). Moreover, sarco-
plasmic protein solubility declines with decreasing pHu and
contributes to paler pork color (Joo et al., 1999). The rate
and extent of postmortem pH decline significantly influence
Fig. 1. Various pH declines occurring postmortem and associated pork
quality characteristics. Modified from Briskey (1964).
 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
protein characteristics and thus critically affect pork quality
development.
3. Major genes and aberrant postmortem metabolism
Postmortem acidification is closely related to the energy
status of the muscle at slaughter. The decline in pH
depends on the initial concentration of PCr and glycogen
(Bendall, 1951), which may vary widely under different
experimental and commercial conditions. A variety of fac-
tors, such as stress, stunning method, muscle type, and diet,
affect muscle energy level and its utilization postmortem.
However, in order to develop more comprehensive and
sophisticated strategies for reducing the variation found
in pork quality, the biochemistry underlying postmortem
metabolism must be fully defined.
3.1. The halothane gene
Early studies utilized certain breeds and lines within
breeds in order to model PSE development. This tendency
to produce PSE pork was closely associated with porcine
stress syndrome, a condition synonymous with human
malignant hyperthermia (MH). Eikelenboom and Mink-
ema (1974) demonstrated that MH susceptible swine dis-
play hypermetabolism, elevated body temperature, and
muscle rigidity upon exposure to the anesthetic halothane.
Stress and excitement are also sufficient to provoke an MH
episode, and can result in the enhanced rate of postmortem
metabolism associated with PSE pork. Exposure to halo-
thane gas was used to screen for stress susceptible swine,
and the genetic component has since been referred to as
the halothane (HAL) gene. The HAL locus has two alleles:
the normal dominant allele (N) and the mutant recessive
allele (n).
MH susceptibility associated with the HAL gene is due
to a defect in the ability of the muscle to adequately regu-
late myoplasmic Ca2+ concentration. The causative poly-
morphism is the single substitution of T for C at
nucleotide 1843 in the skeletal muscle ryanodine receptor
(RYR1), leading to an amino acid change from arginine
to cysteine (Fujii et al., 1991). The RYR1, or Ca2+-release
channel, is the primary mechanism by which Ca2+ stored in
the sarcoplasmic reticulum terminal cisternae is released
into the sarcoplasm to initiate muscle contraction. The
Ca2+ release channels in MH susceptible pigs are hypersen-
sitive to agents that stimulate opening (O’Brien, 1986), thus
allowing longer open time probability and resulting in
enhanced Ca2+ release and greater twitch tension (Mickel-
son et al., 1988; Mickelson et al., 1989). The abnormal
channels flood the cell with Ca2+ and overpowers the sar-
coplasmic reticulum ATPase pumps that resequester cyto-
plasmic Ca2+, resulting in sustained contraction and
metabolism.
This defect in Ca2+ concentration has important conse-
quences for production and meat quality traits. Pigs homo-
zygous for the HAL gene have higher carcass yield and
9
lean percentage (Aalhus, Jones, Robertson, Tong, &
Sather, 1991; Herfort Pedersen et al., 2001; Leach, Ellis,
Sutton, McKeith, & Wilson, 1996; Rempel, Lu, Mickelson,
& Louis, 1995). The leaky Ca2+ release channels of HAL
mutant pigs may contribute to leanness and heavy mus-
cling by causing spontaneous muscle contraction and
greater energy utilization, leading to work induced muscle
hypertrophy and limiting fat deposition (MacLennan &
Phillips, 1992).
The positive effect of the mutant allele on performance is
negated by an increased risk for stress-induced death and
high susceptibility to acute stress prior to slaughter, which
may be manifested in an accelerated rate of pH decline and
the production of PSE pork. An MH episode triggers a
massive stimulation of aerobic and anaerobic metabolism
that depletes energy resources, generates metabolic waste
products, and disrupts cellular and extracellular ion bal-
ances. The larger muscle fiber area and lower capillary den-
sity of homozygous mutant pigs (Essen-Gustavsson,
Karlstrom, & Lundstrom, 1992) further compromises their
ability to cope with metabolic stress, and contributes to an
enhanced rate of glycogenolysis and glycolysis, evidenced
by reduced ATP, PCr, and glycogen levels, and higher lac-
tate levels and lower muscle pH at exsanguination (Essen-
Gustavsson et al., 1992; Fernandez, Neyraud, Astruc, &
Sante, 2002; Lundstrom, Essen-Gustavsson, Rundgren,
Edfors-Lilja, & Malmfors, 1989). Clearly, the mutant
HAL genotype increases the frequency of PSE meat by
accelerating the early postmortem rate of glycogenolysis
and glycolysis, leading to a more rapid pH decline.
3.2. The RN gene
Greater availability of glycogen may contribute to an
extended pH decline by providing additional substrate for
glycolysis. Sayre, Briskey, and Hoekstra (1963) showed
the Hampshire breed had more than twice the amount of
muscle glycogen compared to other breeds. Monin and Sel-
lier (1985) suggested that the higher muscle glycogen stores
were mainly responsible for an extended pH decline post-
mortem, resulting in pale meat with a low pHu, low water
holding capacity, and reduced technological yield, and
referred to this as the ‘‘Hampshire effect’’ in order to distin-
guish it from the PSE meat produced from a rapid pH
decline at elevated temperatures. Monin & Sellier (1985)
recommended that the muscle metabolites glycogen, glu-
cose, glucose 6-phosphate, and lactate should be combined
into a single measure termed glycolytic potential (GP) in
order to reflect all of the compounds in the muscle capable
of being converted to lactate, thus indicating the muscle’s
capacity for extended postmortem glycolysis. A single
point mutation, Rendement Napole (RN), is responsible
for the elevated muscle GP (>180–200 lmol/g muscle),
extended pH decline, and greatly reduced technological
yield that was first found in the Hampshire breed (Le
Roy et al., 2000). There are two alleles, the dominant allele
(RN) associated with elevated muscle glycogen and infe-
10 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
rior quality, and the wild-type recessive allele (rn+)
(Le Roy, Naveau, Elsen, & Sellier, 1990). Additional alleles
in the PRKAG3 gene may play a significant role in glyco-
gen variation, including the 199I allele which is correlated
with lower glycogen and lactate, higher ham and loin pH
and improved color scores, resulting in improved meat
quality (Ciobanu et al., 2001).
The mutation responsible for the RNgene is an R200Q
substitution in the PRKAG3 gene that encodes for the
muscle specific isoform of the regulatory c-subunit of aden-
osine monophosphate activated protein kinase (AMPK)
(Milan et al., 2000). AMPK monitors the energy charge
of the muscle fiber and prevents high energy phosphate
depletion. AMPK is allosterically inhibited by PCr and
0
ATP and activated by 5 AMP. Therefore, increased rates
of ATP utilization during muscle contraction activate
AMPK, which is hypothesized to phosphorylate proteins
involved in triggering fatty acid oxidation and glucose
uptake (reviewed by Winder, 2001). Transfection experi-
ments performed with mice revealed that the PRKAG3
R225Q is a loss of function mutation that eliminates allo-
steric regulation by ATP/AMP resulting in increased basal
AMPK activity (Barnes et al., 2004). Additionally, the
adenosine monophosphate kinase gamma-3 subunit
(AMPKc3) is more highly expressed in mouse fast twitch
white glycolytic muscle compared to fast twitch red oxida-
tive muscle, but is virtually undetectable in red muscle (Yu,
Fujii, Hirshman, Pomerleau, & Goodyear, 2004).
The expression profiling of AMPKc3 supports evidence
that the RN mutation influences the compositional and his-
tochemical traits and metabolic enzyme activities in a mus-
cle-type dependent manner. Lebret et al. (1999) found the
RN mutation had no effect on red semispinalis capitis mus-
cle. However, enhanced glucose uptake due to an increase
in the translocation of GLUT4 to surface membranes
(Kurth-Kraczek, Hirshman, Goodyear, & Winder, 1999)
and faster resynthesis of glycogen after exercise (Anders-
son, 2003) may explain the up to 70% increase in muscle
glycogen localized mostly to abnormally enlarged sarco-
plasmic compartments of type IIB white fibers observed
in RN carriers (Monin, Brard, Vernin, & Naveau, 1992;
Estrade, Vignon, Rock, & Monin, 1993). Furthermore, a
twofold increase in branching enzyme activity and a ten-
dency toward increased glycogen synthase activity may
contribute to elevated glycogen levels and differences in
the molecular structure of glycogen (Estrade, Ayoub, Tal-
mant, & Monin, 1994).
The relatively high glycogen content of RN carriers
might be expected to further enhance glycolytic metabolism
in IIB fibers. Curiously, RN carriers possess enhanced
oxidative metabolism, indicated by higher citrate synthase
and b-hydroxyacyl coenzyme A dehydrogenase activities,
decreased lactate dehydrogenase activity, higher relative
area of IIA fibers and lower relative area of IIB fibers (Leb-
ret et al., 1999). Mice with the PRKAG3 R225Q mutation
also exhibited higher skeletal muscle oxidative capacity
without altered fiber type composition, suggesting that
the elevated glycogen content may be an indirect conse-
quence of altered muscle oxidation, and the role of the
AMPKc3 isoform may be to ensure that glycogen content
in muscle is restored by maintaining a high glycolytic
potential through shifting the metabolic fate of fuel toward
fat oxidation and glycogen storage (Barnes et al., 2004).
Nonetheless, high glycogen levels in RNcarriers do not
appear to alter the rate of glycogen utilization. RNcarri-
ers and wild type pigs exhibit similar glycogen degradation
during exercise (Andersson, 2003) as well as similar rates of
early postmortem pH decline (Monin & Sellier, 1985).
Estrade et al. (1994) reported no differences in either glyco-
gen phosphorylase or debranching enzyme activity, further
supporting that the RN allele increases glycogen avail-
ability without altering the rate of utilization. Thus, the
mutant RN genotype generates pork of inferior technolog-
ical quality by increasing glycolytic potential and extending
postmortem pH decline, resulting in pork with a low pHu.
4. Rate limiting enzymes
Clearly, glycogen degradation through glycolysis
impacts pork quality development. Enzymes catalyzing
the reactions of glycolysis influence the rate and extent of
pH decline by directly controlling the conversion of metab-
olites through the pathway (Fig. 2). The properties of the
rate limiting enzymes glycogen phosphorylase, phospho-
fructokinase, and pyruvate kinase have been studied in
an effort to explain the aberrant glycolysis leading to PSE
development.
Glycogen phosphorylase and glycogen debranching
enzyme catalyze the complete degradation of glycogen.
Glycogen phosphorylase cleaves its substrate by addition
of inorganic phosphate at a-1,4 linkages of the outer chains
of the glycogen molecules, yielding glucose 1-phosphate.
Then, phosphoglucomutase catalyzes the isomerization of
glucose 1-phosphate to glucose 6-phosphate, which can
then proceed through glycolysis. Once phosphorylase
reaches the fourth glucose from the branch point, a trans-
ferase shifts the maltotriosyl group to the main chain, and
glycogen debranching enzyme breaks the a-1,6 linkages,
releasing free glucose. The main chain and remaining outer
branches are again susceptible to breakdown by
phosphorylase.
Sayre et al. (1963) found similar levels of phosphorylase
in swine exhibiting fast and slow rates of glycolysis
postmortem. However, in skeletal muscle, glycogen
phosphorylase exists in two forms: the less active, non-
phosphorylated form (GP b) and the more active, phos-
phorylated form (GP a). Hence, differences in the fraction
of phosphorylase in the a form could plausibly explain the
abnormal glycolysis observed in PSE muscle. Early in vitro
studies indicated a strong relationship between pHu and
the amount of phosphorylase in the a form (Scopes,
1974). In support, Ono, Topel, Christian, and Althen
(1977) demonstrated an enhanced GP a activity in PSE tis-
sue, and Schwagele and Honikel (1988) determined a
 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16 11

Glycogen activity strongly decreased when the temperature decreased

from 39 and 42 °C to 4 and 15 °C. Therefore, the activity of
glycogen phosphorylase debranching enzyme does not block rapid glycolysis and
pH decrease when the temperature is high, but rapid cool-
Glucose Glucose 1 - Phosphate ing could limit the activity and thus limit glycogenolysis.
Phosphofructokinase (PFK) catalyzes the conversion of
hexokinase phosphoglucomutase
fructose 1-phosphate and ATP to fructose 1,6-bisphos-
Glucose 6-Phosphate phate and ADP, which is the committed step in the glyco-
lytic pathway. In an in vitro glycolytic system, PFK was
phosphoglucose isomerase
still active under the extreme PSE conditions of 37 °C
Fructose 6-Phosphate and pH 5.35 (Scopes, 1974). Moreover, total and specific
PFK activities were not significantly different in pigs with
phosphofructokinase (PFK)
45 min longissimus muscle pH values ranging from 5.3 to
Fructose 1,6- Bisphosphate 6.8 (Schwagele & Honikel, 1988). Therefore, activation of
PFK does not readily explain the differences in glycolytic
aldolase
rates between normal and PSE muscles. Interestingly, Alli-
son, Bates, Booren, Johnson, and Doumit (2003) observed
Glyceraldehyde Dihydroxyacetone an inverse relationship between PFK capacity and fluid
3-Phosphate phosphate loss, and reasoned that PFK may become partially dena-
Triose phosphate
isomerase tured and inactivated by 20 min postmortem in muscles
that experience a rapid pH decline. Therefore, it appears
glyceraldehyde 3-phosphate dehydrogenase
that increased PFK activity is not the reason for enhanced
Bisphosphoglycerate glycolysis in PSE muscle.
Pyruvate kinase catalyzes the irreversible conversion of
phosphoglycerate kinase phosphoenolpyruvate and ADP to pyruvate and ATP. Sch-
3-Phosphoglycerate wagele, Haschke, Honikel, and Krauss (1996b) demon-
strated that, compared to normal muscle, pyruvate kinase
phosphoglycerate mutase
isolated from PSE muscle of halothane susceptible pigs
2-Phosphoglycerate exhibited a tenfold increase in phosphoenolpyruvate utili-
zation. Interestingly, the activity of pyruvate kinase from
enolase normal muscle was low at pH 5.5, whereas the enzyme
Phosphenolpyruvate from PSE muscle maintained 70% of its maximum activity
under these acidic conditions. Schwagele et al. (1996b)
pyruvate kinase determined that this shift in pyruvate kinase activity was
Pyruvate due to phosphorylation of the enzyme, resulting in an addi-
tional, more acid stable isoform. It is not clear whether the
lactate dehydrogenase additional, more stable isoform is present in live animals or
Lactate if it is generated under postmortem conditions. Neverthe-
less, in pigs free of the HAL gene, pyruvate kinase capacity
Fig. 2. Enzymes and metabolic intermediates of the glycolytic pathway.
was not correlated with longissimus pH, purge, drip loss or
paleness, and in all samples, pyruvate kinase lost more than
higher total activity (GP a and GP b) in PSE-prone mus- 88% of its activity at pH 5.5 compared to pH 7.0 (Allison
cles. However, Ensinger, Rogdakis, Muller, and Faber et al., 2003). Although the role of pyruvate kinase in HAL
(1982) could not establish differences in the activities of sensitive pigs is not completely understood, it appears that
GP a and GP b in muscles of normal and PSE muscles. enhanced pyruvate kinase capacity is not responsible for
More recently, Schwagele, Buesa, and Honikel (1996a) altered postmortem metabolism in PSE muscle from pigs
reported no significant differences in the structural and free of the HAL gene.
kinetic characteristics of GP a and GP b from muscles of For the most part, the inherent properties of the rate
normal versus HAL sensitive pigs. Thus, the characteristics limiting enzymes are not different in normal versus PSE
of glycogen phosphorylase are not likely to be the main muscles, and thus other mechanisms must be responsible
factors facilitating altered metabolism in PSE muscle. for the aberrant glycogenolysis and glycolysis observed in
Because postmortem glycogenolysis may stop in the PSE muscles. Muscle possesses a complex system of both
presence of residual glycogen, it has been speculated that feedforward and feedback regulation in order to precisely
glycogen debranching enzyme may influence the rate and modulate metabolism, and differences in relative levels of
extent of glycogenolysis and glycolysis. Kyla-Puhju, Ruus- glycolytic regulators could contribute to altered postmor-
unen, and Puolanne (2005) reported that the activity of tem glycolysis. Specifically, the modulators of rate limiting
debranching enzyme was only weakly affected by pH, but enzymes could manipulate enzyme activity according to
12 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
energy demand of the cell, and this would affect flux
through the glycolytic pathway and contribute to altered
postmortem metabolism.
Rapid rates of postmortem glycolysis are associated
with low levels of glycogen and high levels of glucose
6-phosphate, indicative of increased phosphorylase activity
(Briskey et al., 1966; Kastenschmidt, Hoekstra, & Briskey,
1968; Moesgaard, Quistorff, Christensen, Therkelsen, &
Jorgensen, 1995). Glycogen phosphorylase activity is regu-
lated both by phosphorylation and allosteric mechanisms.
Phosphorylase kinase exists in two forms, phosphorylase
kinase a and b (PK a and PK b), which are both capable
of converting GP b to the active GP a form. Maximal acti-
vation of phosphorylase kinase is achieved through phos-
phorylation of PK b to PK a and Ca2+ binding. The
hormones epinephrine and glucagon induce increases in
cAMP that lead to phosphorylation of PK b to PK a,
and PK a is active at the concentration of Ca2+ in resting
muscle. Exposure or susceptibility to stress would permit
epinephrine mediated activation of PK a and subsequent
activation of GP a, resulting in increased glycogenolysis.
The cAMP cascade provides a link between epinephrine
and glycogen degradation, whereas Ca2+ couples muscle
contraction with glycogenolysis (Drummond, Harwood,
& Powell, 1969). Compared to PK a, PK b requires a
higher Ca2+ concentration for activity (Connett & Sahlin,
1996). The Ca2+ concentration in the sarcoplasm during
contraction is sufficient to activate PK b, resulting in con-
version of GP b to the more active GP a form. Preslaughter
stress places an increased demand on muscles for contrac-
tion, and this could also be responsible for increased glyco-
genolysis by mediating the conversion of GP b to GP a by
means of Ca2+. Similarly, in the case of the HAL gene, the
enhanced Ca2+ release may trigger activation of phosphor-
ylase and stimulate glycogenolysis.
The relative concentration of several modulators related
to cellular energy charge and substrate availability regulate
the activity of GP b. The levels of inhibitors, including
ATP, ADP, and glucose 6-phosphate, are usually sufficient
to inhibit GP b in resting muscle, but their effect may be
overcome by increases in AMP and inosine monophos-
phate (IMP), and lead to GP b activation (Connett & Sah-
lin, 1996). Furthermore, the sensitivity of GP b for each
factor is dependent on the concentration of substrate (gly-
cogen and inorganic phosphate) and product (glucose 1-
phosphate). In contrast, GP a is active in the absence of
AMP, although activity is enhanced by low concentrations
of AMP and high concentrations of IMP. The other allo-
steric modulators have little effect on GP a activity.
In muscles exhibiting normal rates of glycolysis, glucose
6-phosphate levels tend to decrease during the first hour
postmortem and increase thereafter (Kastenschmidt et al.,
1968; Hammelman et al., 2003). This suggests that glyco-
gen phosphorylase may be unable to supply adequate glu-
cose 6-phosphate for the subsequent reactions of the
glycolytic pathway, resulting in a rapid utilization of the
glucose 6-phosphate pool. If animals exhibiting ‘‘slow’’
rates of glycolysis were assumed to experience minimal
antemortem stress, glycogen phosphorylase would exist
predominantly in the b form. Moreover, following exsan-
guination, the relatively high levels of ATP and low levels
of AMP (Kastenschmidt et al., 1968) and IMP (Klont &
Lambooy, 1995) would be insufficient to activate GP b.
After the first hour postmortem, changes in the relative
concentrations of allosteric activators may permit GP b
activation. Therefore, the accumulation of G6P after one
hour may be the result of increased glycogen breakdown
due to glycogen phosphorylase, or a decrease in the activity
of PFK or other enzymes downstream.
The regulation of the other rate limiting enzyme, PFK is
less complex. Similar to GP b, PFK activity depends largely
on the energy status of the muscle cell. As the ATP/AMP
ratio decreases, PFK activity is stimulated. PFK is also
activated by hexose bisphosphates and inorganic phos-
phate. Moreover, ATP is required for the transfer of a
phosphate group from ATP to fructose 6-phosphate.
Rapid depletion of ATP in fast glycolyzing muscles (Kas-
tenschmidt et al., 1968) could compromise the ability of
PFK to catalyze the formation of fructose 1,6-bisphos-
phate. This would suggest PFK is exerting some glycolytic
control. However, in the case of slow glycolyzing muscles,
the imbalance between fructose 6-phosphate and fructose
1,6-bisphosphate becomes apparent after 60 min, and Kas-
tenschmidt et al. (1968) suggested this may be due to
decreasing muscle pH and partial inactivation of PFK.
In contrast, flux through pyruvate kinase appears to be
largely governed by the concentration of the substrates
phosphoenolpyruvate and ADP and product ATP. The
high levels of ATP present in slow glycolyzing muscles
may exert some control over PK early postmortem. How-
ever, in fast glycolyzing muscles, lactate production ceased
in the presence of residual glycogen, glucose 1-phosphate,
glucose 6-phosphate (G6P), and fructose 6-phosphate,
whereas subsequent intermediates were present at low lev-
els (Kastenschmidt et al., 1968). This implies PFK is more
likely than PK to be a site of glycolytic control.
Altogether, previous studies suggest that the relative lev-
els of glycolytic regulators contribute to altered postmor-
tem glycolysis. Specifically, the modulators of rate
limiting enzymes manipulate enzyme activity according to
cellular energy status, and this affects flux through the
pathway. Glucose 6-phosphate levels fall during the first
60 min, and subsequently increase, indicating an imbalance
between glycogenolysis and glycolysis. Therefore, different
enzymes may be rate limiting at different times during the
conversion of muscle to meat.
5. Control of postmortem glycolysis by AMPK
AMPK is particularly attractive for augmenting post-
mortem metabolism primarily because in vivo studies show
AMPK is activated in ischemic cardiac muscle and hypoxic
skeletal muscle (Kim, Solis, & Cartee, 2004). As outlined
above, when muscle (cardiac or skeletal) is placed in a
 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
low oxygen environment, the primary source of energy pro-
duction defaults to anaerobic glycolysis, thereby increasing
the AMP:ATP ratio rapidly. This, in turn, allows AMP to
bind to AMPK making it a better substrate for phosphor-
ylation by its upstream kinase, AMPKK. Activated
AMPK can impact glycolysis in two manners. First,
AMPK can activate phosphorylase kinase, which then
activates glycogen phosphorylase and promotes glycogen-
olysis. Second, AMPK can phosphorylate phosphofructo-
kinase-2 which catalyzes the formation of fructose
2,6-bisphosphate. This product is an allosteric activator
of PFK-1, a key rate-limiting enzyme of glycolysis. Thus,
AMPK activation indirectly increases flux through glycol-
ysis making it a strong candidate for modulating meat
quality development.
Shen and Du (2005) first investigated the involvement of
AMPK in regulating, or modulating postmortem metabo-
lism by studying exercised wild-type and AMPK-knockout
mice treated with or without the AMP analog 5-amino-4-
imidazolecarboxamide riboside (AICAR). These scientists
showed heavily exercised mice and those exercised mice
treated with AICAR had significantly lower (more rapid)
muscle pH values at 24 h postmortem (Fig. 3). Conversely,
mice lacking a functional AMPK gene, whether exercised
or not, had higher ultimate muscle pH values, even higher
than wild-type, non-exercised mice. These data closely
reflected trends in AMPK activity suggesting that aug-
mented postmortem glycolysis in mouse skeletal muscle is
partially controlled by the status of AMPK phosphoryla-
tion antemortem. To eliminate the effects of AMPK activa-
tion in living muscle on postmortem metabolism, Du et al.
(personal communication) infused muscle with compound
C immediately antemortem. In good agreement with their
aforementioned data, postmortem glycolysis and muscle
pH decline was inhibited. These data strongly support the
role of AMPK in modulating postmortem muscle
metabolism.
At a glance, these data appeared to shed much light on
the 50 year debate as to what may be controlling extended
postmortem glycolysis in pig muscle. However, conditions
whereby mouse and pig muscle undergo postmortem
Fig. 3. Muscle pH values at various times postmortem from mice treated
differently prior to euthanasia (Shen & Du, 2005).
13
metabolism are quite different. To that end, this same
group (Shen et al., 2006a) used pre-slaughter transporta-
tion as a stressor and monitored AMPK activity and
energy status of pig muscle postmortem. Similar to the
mouse data, AMPK activity reached maximum levels
quicker in transported pig muscle than in control pigs, or
those rested after transport. Consistent with these findings,
the (AMP + IMP):ATP ratio was greater in the trans-
ported pigs arguing AMPK activation through cytosolic
AMP concentrations. These researchers subsequently
showed an association between early postmortem AMPK
activation and PSE development in commercial pigs and
suggested that the rapid glycolysis occurring in muscle of
HAL pigs was in part from increases in AMPK activation
early postmortem (Shen et al., 2006b). These data clearly
show an association between AMPK activation and aggra-
vated metabolism postmortem. However, much more work
is needed to understand exactly how AMPK works in
dying muscle tissue.
We have attempted to study muscle metabolism further
using the HAL and RN mutations separately and in com-
bination – rapid metabolism together with abundant gly-
cogen supply (HAL/RN mutant) – to provide additional
insight into the control of glycogenolysis and glycolysis
(Copenhafer, Richert, Schinckel, Grant, & Gerrard,
2006). As expected, the previously documented effects of
both genes were evident: the HAL mutation elicited has-
tened metabolism, as evidenced by lower ATP levels at
0 and 30 min, and rapid degradation of glycogen and
accumulation of lactate early postmortem compared to
control; whereas the RN mutation resulted in much
greater glycolytic potential due to elevated glycogen
levels.
Additional investigation yielded more insight regarding
biochemical events within the glycolytic pathway. Free glu-
cose, released by the action of debranching enzyme on gly-
cogen, was independently affected by HAL and RN
genotype. The HAL mutants exhibited increased glucose
accumulation indicative of more rapid glycogen degrada-
tion and early postmortem metabolism, whereas the
increases in the RN mutants occurred to a greater extent
and after 60 min postmortem. Rapid rates of glycolysis
were associated with decreased glycogen as well as high
concentrations of glucose 6-phosphate, indicating
increased phosphorylase activity. G6P concentrations
decreased in the HAL mutants during the first 30 min
and remained at a similar level thereafter. Meanwhile, in
the HAL/RN mutant, G6P concentrations tended to
increase in the first 30 min, then decrease through 60 min,
indicating the combined effects of the HAL and RN max-
imize activation of glycogen phosphorylase by enhancing
Ca2+ concentration and glycogen availability. Despite large
differences in G6P between HAL and HAL/RN mutants,
pH decline and lactate accumulation were similar early
postmortem. Glycogen phosphorylase and debranching
enzyme were capable of aggressive glycogenolysis to supply
adequate G6P for the reactions of glycolysis. Additionally,
14 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
high glycogen levels did not aggravate rapid postmortem
metabolism.
From 60 min to 24 h postmortem, G6P increased only in
normal and RN mutant genotypes. The accumulation of
G6P after 60 min confirmed that glycogen phosphorylase
activity was adequate to meet the demands of glycolysis
and that either glycogen phosphorylase activity was
increased or phosphofructokinase activity was reduced.
Similar residual glycogen levels as well as the slowing of
glycolysis with time, indicated that increased G6P is likely
due to reduced phosphofructokinase activity.
Interestingly, RN mutants possessed higher phosphocre-
atine concentrations than all other genotypes (Fig. 4).
Ponticos et al. (1998) observed that a fall in PCr:creatine
ratio leads to increased AMPK activity, which results in
a concomitant decrease in muscle CK activity. The pro-
posed inactivation of CK by AMPK may be an energy con-
servation mechanism that prevents CK from consuming
ATP for the rephosphorylation of creatine. Presence of
the mutant RN genotype also resulted in an increase in
ATP concentrations. The constitutively active AMPK in
the RNmutant genotype appears to decrease CK activity
and preserve energy levels more efficiently in the first
30 min postmortem. This may have several ramifications
in extending postmortem proteolysis. First, it could pro-
long glycolysis by having greater adenosine levels in the tis-
sue longer. Second, this could delay maximal rate of
glycolysis, which would likely occur at a reduced carcass
temperature allowing for extended glycolysis. And third,
the increase in inorganic phosphate may increase the buf-
fering capacity of the muscle, and again, allow for extended
glycolysis.
The RN allele is typically associated with increased
glycolytic potential and lower ultimate pH. However, at
24 h postmortem, RN and HAL/RN mutant pigs exhibited
12
Normal
RN
10 a
HAL
HAL/RN
Phosphocreatine (umol/g)
8 by the fact that we observed greater glycolytic potential
b Given that increased GLUT-4 production, the primary
6
b b
b
4 how greater glycogen is accumulated in the muscle of
c HAL/RN mutants. At present, the only possibility is that
c c
2
0 Taken together, the aforementioned review shows that
0 min 30 min
Time
Fig. 4. LS means of postmortem longissimus muscle phosphocreatine
concentrations in halothane and RN genotypes. (Copenhafer et al., 2006).
lower pH than the normal genotype but not the HAL
mutants. Previous studies have also indicated that the
mutant HAL allele tends to lower ultimate pH. Therefore,
substrate availability alone does not explain variation in
ultimate pH. Data from this study show that elevated gly-
cogen does not aggravate rapid early postmortem glycoly-
sis, nor does it fully explain low ultimate pH. The
diminished ATP in muscles undergoing rapid glycolysis
suggests phosphofructokinase may be rate limiting during
aggressive early postmortem metabolism. Moreover, the
accumulation of glucose and glucose 6-phosphate in mus-
cles with normal rates of glycolysis suggests phosphofruc-
tokinase may be rate limiting after 1 h postmortem.
Thus, depending on the pace of postmortem metabolism,
enzymes may become rate limiting at different times during
postmortem metabolism. Regardless, this is a truly valu-
able model for studying adverse pork quality development.
Further analyses of muscle tissues from the aforemen-
tioned study showed that AMPK activation was greater
in muscle from RNpigs compared to all other genotypes
(Park et al., submitted). Curiously, classic data from Milan
et al. (2000) reported that AMPK activation in RN-pigs is
greatly reduced compared to wild type pigs. The reason for
this discrepancy is not readily apparent but may be related
to sampling time or muscle type. In our studies, samples
were collected immediately post-stunning from the longiss-
imus muscle, however, details regarding sampling proce-
dures in the Milan study are somewhat sketchy. Our
data, however, are corroborated by data from the AMPK
RN mutant mice, where increased AMPK activation is
observed in that muscle as well. Clearly, establishing
whether AMPK is activated in RN mutated pig muscle,
in either living or dying muscle is critical for our under-
standing of how AMPK may modulate postmortem muscle
metabolism.
Another intriguing observation merits discussion is that
AMPK activation is blunted in the presence of the HAL
gene (Park et al., submitted). Thus, if it is true that AMPK
modulates postmortem energy metabolism as reported by
Shen et al. (2006), then the state of AMPK prior to slaugh-
ter may be more important than what happens to the
enzyme after death. This scenario is further complicated
in the double mutant, in the absence of AMPK activation.
means of glucose uptake in the muscle, is only observed
in muscle with activated AMPK, it is difficult to imagine
phosphorylation of AMPK may not be the only mecha-
nism by which AMPK modulates energy balance in muscle.
Additional studies are needed to support this hypothesis.
postmortem metabolism in skeletal muscle is quite compli-
cated and largely affected by the state of the tissue prior to
death. Regardless of the complexity, however, numerous
genetic models, molecular techniques and commercially
 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
available compounds exist that make studying this event
quite rewarding. If technologies are going to be developed
that allow the pork industry to predict quality, then a com-
prehensive understanding of the biochemical and molecu-
lar events controlling postmortem metabolism must be
known.
References
Aalhus, J. L., Jones, S. D. M., Robertson, W. M., Tong, A. K. W., &
Sather, A. P. (1991). Growth characteristics and carcass composition
of pigs with known genotypes for stress susceptibility over a weight
range of 70 to 120 kg. Animal Production, 52, 347–353.
Allison, C. P., Bates, R. O., Booren, A. M., Johnson, R. C., & Doumit, M.
E. (2003). Pork quality variation is not explained by glycolytic enzyme
capacity. Meat Science, 63, 17–22.
Andersson, L. (2003). Identification and characterization of AMPK
gamma 3 mutations in the pig. Biochemical Society Transactions, 31,
232–235.
Barnes, B. R., Marklund, S., Steiler, T. L., Walter, M., Hjalm, Goran,
Amarger, V., et al. (2004). The 5’-AMP-activated protein kinase
gamma 3 isoform has a key role in carbohydrate and lipid metabolism
in glycolytic skeletal muscle. Journal of Biological Chemistry, 279,
38441–38447.
Bate-Smith, E. C., & Bendall, J. R. (1949). Factors determining the time
course of rigor mortis. Journal of Physiology, 110, 47–65.
Bendall, J. R. (1951). The shortening of rabbit muscles during rigor
mortis: Its relation to the breakdown of adenosine triphosphate and
creatine phosphate and to muscular contraction. Journal of Physiology,
114, 71–88.
Bendall, J. R. (1973). Post mortem changes in muscle. In G. H. Bournes
(Ed.), The structure and function of muscle (pp. 227–274). New York:
Academic Press.
Briskey, E. J. (1964). Etiological status and associated studies of pale, soft,
exudative porcine musculature. Advances in Food Research, 13, 89–178.
Briskey, E. J., Kastenschmidt, L. L., Forrest, J. C., Beecher, G. R., Judge,
M. D., Cassens, R. G., et al. (1966). Biochemical aspects of post-
mortem changes in porcine muscle. Journal of Agricultural and Food
Chemistry, 14, 201–207.
Briskey, E. J., & Wismer-Pedersen, J. (1961). Biochemistry of pork muscle
structure. I. Rate of anaerobic glycolysis and temperature change
versus the apparent structure of muscle tissue. Journal of Food Science,
26, 297–305.
Carr, T. R., Kauffman, R. G., Meeker, D. L., & Meisinger, D. L. (1997).
Factors reducing pork value Pork Industry Handbook, PIH-135. West
Lafayette, IN: Purdue University Cooperative Extension Service, pp.
1–4.
Cassens, R. G. (2000). Historical perspectives and current aspects of pork
meat quality in the USA. Food Chemistry, 69, 357–363.
Ciobanu, D., Bastiaansen, J., Malek, M., Helm, J., Woollard, J., Plastow,
G., et al. (2001). Evidence for new alleles in the protein kinase
adenosine monophosphate-activated gamma3-subunit gene associated
with low glycogen content in pig skeletal muscle and improved meat
quality. Genetics, 159, 1151–1162.
Connett, R. J., & Sahlin, K. (1996). Control of glycolysis and glycogen
metabolism. In L. B. Rowell & J. T. Shepherds (Eds.), Handbook of
physiology. Exercise: Regulation and integration of multiple systems
(pp. 871–911). New York: Oxford University Press.
Copenhafer, T. L., Richert, B. T., Schinckel, A. P., Grant, A. L., &
Gerrard, D. E. (2006). Augmented postmortem glycolysis does not
occur early postmortem in AMPKc3-mutated porcine muscle of
Halothane positive pigs. Meat Science, 73, 590–599.
Drummond, G. I., Harwood, J. P., & Powell, C. A. (1969). Studies on
the activation of phosphorylase in skeletal muscle by contraction
and by epinephrine. Journal of Biological Chemistry, 244,
4235–4240.
15
Eikelenboom, G., & Minkema, D. (1974). Prediction of pale, soft,
exudative muscle with a non-lethal test for the halothane-induced
porcine malignant hyperthermia syndrome. Tijdschr Diergeneesk, 99,
421–426.
Ensinger, V. U., Rogdakis, E., Muller, E., & Faber, H. (1982). Parameters
of metabolism in halothane-negative pigs. I. Concentration of lactate,
cAMP, glycogen, and activity of phosphorylase, phosphofructokinase,
and pyruvate kinase in the m. Longissimus dorsi. Zeitschrift fur
Tierzuchtung und Zuchtungsbiologie, 99, 26–32.
Essen-Gustavsson, B., Karlstrom, K., & Lundstrom, K. (1992). Muscle
fibre characteristics and metabolic response at slaughter in pigs of
different halothane genotypes and their relation to meat quality. Meat
Science, 31, 1–11.
Estrade, M., Ayoub, S., Talmant, A., & Monin, G. (1994). Enzyme
activities of glycogen metabolism and mitochondrial characteristics in
muscles of RN-carrier pigs (Sus scrofa domesticus). Comparative
Biochemistry and Physiology Part B: Biochemistry and Molecular
Biology, 108, 295–301.
Estrade, M., Vignon, X., Rock, E., & Monin, G. (1993). Glycogen
hyperaccumulation in white muscle fibres of RN-carrier pigs. A
biochemical and ultrastructural study. Comparative Biochemistry and
Physiology Part B: Biochemistry and Molecular Biology, 104, 321–326.
Fernandez, X., Neyraud, E., Astruc, T., & Sante, V. (2002). Effects of
halothane genotype and pre-slaughter treatment on pig meat quality.
Part 1. Post mortem metabolism, meat quality indicators and sensory
traits of m. Longissimus lumborum. Meat Science, 62, 429–437.
Fujii, J., Otsu, K., Zorzato, F., de Leon, S., Khanna, V. K., Weiler, J. E.,
et al. (1991). Identification of a mutation in porcine ryanodine
receptor associated with malignant hyperthermia. Science, 253,
448–450.
Hammelman, J. E., Bowker, B. C., Grant, A. L., Forrest, J. C., Schinckel,
A. P., & Gerrard, D. E. (2003). Early postmortem electrical stimula-
tion simulates PSE pork development. Meat Science, 63, 69–77.
Herfort Pedersen, P., Oksbjerg, N., Karlsson, A. H., Busk, H., Bendixen,
E., & Henckel, P. (2001). A within litter comparison of muscle fibre
characteristics and growth of halothane carrier and halothane free
crossbreed pigs. Livestock Production Science, 73, 15–24.
Honikel, K. O., & Kim, C.-J. (1986). Causes of the development of PSE
pork. Fleischwirtsch., 66, 349–353.
Irving, T. C., Swatland, H. J., & Millman, B. M. (1989). X-ray diffraction
measurements of myofilament lattice spacing and optical measure-
ments of reflectance and sarcomere length in commercial pork loins.
Journal of Animal Science, 67, 152–156.
Joo, S. T., Kauffman, R. G., Kim, B. C., & Park, G. B. (1999). The
relationship of sarcoplasmic and myofibrillar protein solubility to
colour and water-holding capacity in porcine longissimus muscle.
Meat Science, 52, 291–297.
Kastenschmidt, L. L., Hoekstra, W. G., & Briskey, E. J. (1968). Glycolytic
intermediates and co-factors in ‘‘fast-’’ and slow-glycolyzing’’ muscles
in pig. Journal of Food Science, 33, 151–158.
Kim, J., Solis, R.S. Arias, E.B. & Cartee, G.D. (2004) Postcontraction
insulin sensitivity: relationship with contraction protocol, glycogen
concentiration and 5’ AMP-activiated protein kinase phosphorylation.
Journal of Applied Physiology. 96, 575–583.
Klont, R. E., & Lambooy, E. (1995). Influence of preslaughter muscle
temperature on muscle metabolism and meat quality in anesthetized
pigs of different halothane genotypes. Journal of Animal Science, 73,
96–107.
Kurth-Kraczek, E. J., Hirshman, M. F., Goodyear, L. J., & Winder, W.
W. (1999). 5’ AMP-activated protein kinase activation causes GLUT4
translocation in skeletal muscle. Diabetes, 48, 1667–1671.
Kyla-Puhju, M., Ruusunen, M., & Puolanne, E. (2005). Activity of
porcine muscle glycogen debranching enzyme in relation to pH and
temperature. Meat Science, 69, 143–149.
Leach, L. M., Ellis, M., Sutton, D. S., McKeith, F. K., & Wilson, E. R.
(1996). The growth performance, carcass characteristics, and meat
quality of halothane carrier and negative pigs. Journal of Animal
Science, 74, 934–943.
16 T.L. Scheffler, D.E. Gerrard / Meat Science 77 (2007) 7–16
Lebret, B., Le Roy, P., Monin, G., Lefaucheur, L., Caritez, J. C., Talmant,
A., et al. (1999). Influence of the three RN genotypes on chemical
composition, enzyme activities, and myofiber characteristics of porcine
skeletal muscle. Journal of Animal Science, 77, 1482–1489.
Le Roy, P., Elsen, J. M., Caritez, J. C., Talmant, A., Juin, H., Sellier, P.,
et al. (2000). Comparison between the three porcine RN genotypes for
growth, carcass composition and meat quality traits. Genetics Selection
Evolution, 32, 165–186.
Le Roy, P., Naveau, J., Elsen, J. M., & Sellier, P. (1990). Evidence for a
new major gene influencing meat quality in pigs. Genetical Research,
55, 33–40.
Lundstrom, K., Essen-Gustavsson, B., Rundgren, M., Edfors-Lilja, I., &
Malmfors, G. (1989). Effect of halothane genotype on muscle
metabolism at slaughter and its relationship with meat quality: A
within litter comparison. Meat Science, 25, 251–263.
MacLennan, D. H., & Phillips, M. S. (1992). Malignant hyperthermia.
Science, 256, 789–794.
Mickelson, J., Gallant, E., Litterer, L., Johnson, K., Rempel, W., & Louis,
C. (1988). Abnormal sarcoplasmic reticulum ryanodine receptor in
malignant hyperthermia. Journal of Biological Chemistry, 263,
9310–9315.
Mickelson, J. R., Gallant, E. M., Rempel, W. E., Johnson, K. M., Litterer,
L. A., Jacobson, B. A., et al. (1989). Effects of the halothane sensitivity
gene on sarcoplasmic reticulum function. American Journal of Phys-
iology – Cell Physiology, 26, C787–C794.
Milan, D., Jeon, J. T., Looft, C., Amarger, V., Robic, A., Thelander, M.,
et al. (2000). A mutation in PRKAG3 associated with excess glycogen
content in pig skeletal muscle. Science, 288, 1248–1251.
Moesgaard, B., Quistorff, B., Christensen, V. G., Therkelsen, I., &
Jorgensen, P. F. (1995). Differences of post-mortem ATP turnover in
skeletal muscle of normal and heterozygote malignant-hyperthermia
pigs: Comparison of 31P-NMR and analytical biochemical measure-
ments. Meat Science, 39, 43–57.
Monin, G., Brard, C., Vernin, P., & Naveau, J. (1992). Effects of the RN-
gene on some traits of muscle and liver in pigs. In: 38th ICoMST,
Clermont-Ferrand, France. pp. 391–394.
Monin, G., & Sellier, P. (1985). Pork of low technological quality with a
normal rate of muscle pH fall in the immediate post-mortem period:
The case of the Hampshire breed. Meat Science, 13, 49–63.
O’Brien, P. J. (1986). Porcine malignant hyperthermia susceptibility:
hypersensitive calcium release mechanism of skeletal muscle sarco-
plasmic reticulum. Canadian Journal of Veterinary Research, 50,
318–328.
Offer, G., Knight, P. K., Jeacocke, R., Almond, R., Cousins, T., Elsey, J.,
et al. (1989). The structural basis of the water-holding, appearance
and toughness of meat and meat products. Food Microstructure, 8,
151–170.
Ono, K., Topel, D. G., Christian, L. L., & Althen, T. G. (1977).
Relationship of cyclic-AMP and phosphorylase a in stress susceptible
and control pigs. Journal of Food Science, 42, 108–110.
Ponticos, M., Lu, Q. L., Morgan, J. E., Hardie, D. G., Partridge, T. A., &
Carling, D. (1998). Dual regulation of the AMP-activated protein
kinase provides a novel mechanism for the control of creatine kinase in
skeletal muscle. EMBO Journal, 17, 1688–1699.
Rempel, W. E., Lu, M. Y., Mickelson, J. R., & Louis, C. F. (1995). The
effect of skeletal muscle ryanodine receptor genotype on pig perfor-
mance and carcass quality traits. Animal Science, 60, 249–257.
Sayre, R. N., Briskey, E. J., & Hoekstra, W. G. (1963). Comparison of
muscle characteristics and post-mortem glycolysis in three breeds of
swine. Journal of Animal Science, 22, 1012–1020.
Schwagele, F., & Honikel, K. O. (1988). Studies in postmortem metab-
olism of PSE-prone pork muscles. In: 34th International Congress of
Meat Science and Technology, Brisbane, Australia.
Schwagele, F., Buesa, P. L. L., & Honikel, K. O. (1996a). Enzymological
investigations on the causes for the PSE-syndrome, II. Comparative
studies on glycogen phosphorylase from pig muscles. Meat Science, 44,
41–53.
Schwagele, F., Haschke, C., Honikel, K. O., & Krauss, G. (1996b).
Enzymological investigations on the causes for the PSE-syndrome, I.
Comparative studies on pyruvate kinase from PSE- and normal pig
muscles. Meat Science, 44, 27–40.
Scopes, R. K. (1974). Studies with a reconstituted muscle glycolytic
system. The rate and extent of glycolysis in simulated postmortem
conditions. Biochemical Journal, 142, 79–86.
Sellier, P., & Monin, G. (1994). Genetics of pig meat quality: A review.
Journal of Muscle Foods, 5, 187–219.
Shen, Q. W., & Du, M. (2005). Role of AMP-activated protein kinase in
the glycolysis of postmortem muscle. Journal of the Science of Food and
Agriculture, 85, 2401–2406.
Shen, Q. W., Means, W. J., Thompson, S. A., Underwood, K. R., Zhu, M.
J., McCormick, R. J., et al. (2006a). Pre-slaughter transport, AMP-
activated protein kinase, glycolysis, and quality of pork loin. Meat
Science, 74, 388–395.
Shen, Q. W., Means, W. J., Underwood, K. R., Thompson, S. A., Zhu, M.
J., McCormick, R. J., et al. (2006b). Early post-mortem AMP-
activated protein kinase (AMPK) activation leads to phosphofructo-
kinase-2 and -1 (PFK-2 and PFK-1) phosphorylation and the
development of pale, soft, and exudative (pse) conditions in porcine
longissimus muscle. Journal of Agricultural and Food Chemistry, 54,
5583–5589.
Stetzer, A. J., & McKeith, F. K. (2003). Benchmarking value in the pork
supply chain: Quantitative strategies and opportunities to improve
quality. Savoy, IL: American Meat Science Association.
Winder, W. W. (2001). Energy-sensing and signaling by AMP-activated
protein kinase in skeletal muscle. Journal of Applied Physiology, 91,
1017–1028.
Yu, H., Fujii, N., Hirshman, M. F., Pomerleau, J. M., & Goodyear, L. J.
(2004). Cloning and characterization of mouse 5’-AMP-activated
protein kinase gamma3 subunit. American Journal of Physiology – Cell
Physiology, 286, C283–C292.