The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series Year II (XXXI)

EFFECT OF PAPAIN AND BROMELIN ON MUSCLE AND COLLAGEN

PROTEINS IN BEEF MEAT
Aurelia IONESCU, Iuliana APRODU, Gina PASCARU
Department of Biochemistry, Dunarea de Jos University of Galati, Faculty of Food Science and Engineering,
111, Domeasca St., Tel./Fax: +40 236 460165

Received 28 May - Accepted 9 July

Abstract
The effects of papain and bromelin on muscles and collagen proteins in beef meat were evaluated by
injecting brine supplied with enzymes in different concentrations. The effects produced by papain and
bromelin, together with the endogenous enzymes of meat, were established by the determination of
nitrogenous compounds resulted through the degradation of meat proteins and of hydroxyproline
produced from collagen during enzymatic tenderization and thermal treatment of the meat. Papain and
bromelin led to a limited hydrolysis of beef meat proteins, to a loss of physical integrity of muscle and
connective tissue, accompanied by a high solubility of structural proteins, and to an improvement of the
beef meat tenderness. The tenderization effects of papain and bromelin were monitorized, both on the raw
meat during ageing at 4oC for 24 and 48 hours and on the cooked meat.
Keywords: papain, bromelin, myofibrillar proteins, collagen, rigidity index, tenderness

enzymes of vegetable
origin, such as papain,
(Koohmaraie,
1992a, bromelin and ficin and
1. Introduction
1992b). In the USA, beef bacterial
collagenase
meat is classified into (Foegeding and Larick,
Tenderness is one of most important meatgroups,
based
on 1986; Kang and Rice,
texture attributes which affects the perception of different
levels
of 1970; Stanton and Light,
beef meat, by the customers (Brooks et al., palatability. Between the
1987) were often used for
2000; Morgan et al., 1991). Tenderness dependsmarbling
and
the postmortem
meat
on the structural integrity of the myofibrils and palatability of beef meat,
tenderization,
without
of connective tissue which surrounds the muscle there
is
a
direct
exactly,
fibers and on their properties (Bailey, 1972; correlation: meats with a establishing
which
enzyme
is
more
Koohmaraie et al., 1988; Koohmaraie, 1994;high level of marbling
Nishimura et al., 1996a, 1996b; Penny, 1980).are juicier and tenderer. efficient. These enzymes
Collagen plays an important role in the meatIn the last years different have a large spectrum of
texture. The contribution of connective tissue to tenderization techniques action being involved in
the secondary toughness of meat is dependent of sheep and beef meat degradation of the main
on the quantity, type and intermolecular cross- were applied. These proteins of myofibrillar
muscles
(myofibrillar
links of collagen which is the main component
techniques
include proteins, collagen), and
of connective tissue (Bailey and Light, 1989;
mechanical tenderization, sometimes lead to over
Bertran et al., 1992; Light et al., 1985). By
storage
at
high tenderization and to a
formation of the cross-links between the
temperatures, injection product with a pasty
collagen molecules, the meat of old animals
with calcium chloride, texture (Miller, 1995).
becomes harder. In order to obtain tender meat
electrical
stimulation, Nowadays there is not
at industrial level, it proceeds either to the
ultrasonation,
high clear evidence which
slaughter of young animals, or to the storage of
pressure
treatment
and
to
collagen
meats for ageing at low temperatures (0-4 oC).enzymatic tenderization refers
degradation
during
postDuring the ageing it takes place a limited(Cheftel and Culioli,
process of proteolysis which leads to1997; Koohmaraie et al.,
ultrastructural changes in skeletal muscle and to1988;
Koohmaraie,
the improvement of meat tenderness
1992a,
1992b).
The
Corresponding

author:

aurelia.ionescu@ugal.

ro

Ionescu et.al . / The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series, II (XXXI), 2008, 9-16

currently used in
mortem ageing ofmeat industry.
beef meat, but only toWe tested the
the
structuralfollowing
weakening of theenzymes:
endomysium
andbromelin
[EC
perimysium
during3.4.4.24]
and
the ageing for longpapain
[EC
periods
of
time3.4.4.10],
(Nishimura et al.,purchased from
1998;
Takahashi,Difco, N.F.VIII
1996). Phillips et al.
(Difco
(2000)
tried
to
Laboratories,
establish the way in
Detroit,
which the disruption
Michigan).
of connective tissue
matrix contributes to2.2. Chemical
the improvement ofanalysis
tenderness.
Moisture content
The present study was determined
aimed
at
the by drying the
evaluating
papain sample for 24
and bromelin effects hours at 100oC,
on the beef meat according
to
proteins,
at method AOAC,
determining
the 1995.
optimal quantity and Total
nitrogen
time of enzyme content
was
action
and
at determined
estimating of the according to the
effect of thermal and method AOAC,
enzymatic treatment 1995.
The
on the beef meat content of global
tenderization.
proteins
was
established
by
2. Materials and
multiplying the
Methods
percentage of the
total
nitrogen
2.1. Materials
content by 6.25
The
commercially proteins.
farmed
beef
of Fat content was
approximately
24 determined
months of age were according to the
taken from a single method AOAC,
commercial producer 1984 which is
and slaughtered at an based on the
EU approved abattoir extraction of free
using
standard fat
substances
international
from
analyzed
procedures.
sample
with
The
salt,
tripolyphosphate and
sugar used for the
study
are
food
additives which are

ethylic ether.
The
measurement of
the pH was made
according to the

procedure
AOAC,
1984;
10g of sample
were
homogenized for
2 minutes with
90
ml
of
distillated water
using
a
laboratory
blender; the meat
suspension was
filtrated and the
pH
was
determined with
a pH- meter
digital Hanna.

2.3. Level of
hydrolysis

for
the sample
determination of deformation when
a force is applied
The partial hydrolysisthe amino acids.
perpendicular to
level of the beefSoluble proteins,
muscle
fibers
muscle proteins by thefrom the juice
(Voisey,
1976).
exogenous
enzymesexpressed during Rigidity
index
was appreciated bythe boiling of the (Ir) was estimated
determination of theenzymatic treated through
the
were
following
nitrogenmeat,
relation:
determined
fractions:
Ir=a/g
through
the
Non-protein nitrogen colorimetric
[cm2/g]
- The fraction of non-method of Lowry,
where: a = the
protein nitrogen wasmodified
by surface occupied
determined
afterMiller.
Soluble by meat after
protein precipitationproteins
were pressing it with a
with trichloroaceticexpressed
as weight of 1kg for
2
acid and removing itbovine
serum 10 minutes, cm ,
by
filtration,albumin.
while g = the
according
to
the
mass of meat to
method AOAC, 1990.Solubilization of be pressed, g.
The filtrate free ofcollagen - The
proteins
wassolubilization of 2.4. Preparation
was of samples
mineralized,
withcollagen
measured
by
concentrated sulfuric
The beef ham (24
the
acid in presence ofdetermining
hours
after
catalysts; the contentfree
slaughter)
was
of
nitrogen
washydroxyproline
chosen from fats
determined from thefrom the juice and
connective
was
mineralized samplewhich
tissues and was
after dilution andeliminated at the divided in equal
alkalization by theboiling of the pieces in length
enzymatic treated
micro-Kjeldahl
and
thickness,
samples, according having
method.
the
to the method approximate weigh
Aminic nitrogen - The
Standard
ISO of 100 g. The
free amino acids,
3496/1997.
cutting of pieces
expressed as tyrosine,
Tenderization
was made along
were dosed from the
level
was
the muscle fibers.
extract
free
of
appreciated
by
The pieces of meat
proteins
and
determination
of
were divided into
polypeptides
the
rigidity
index.
4 groups; each
insoluble
in
Compression
group, having at
trichloroacetic
acid
least 5 pieces, was
method
is
based
solution, using the
in
a
recording treated
colorimetric methodon
different way:
10

The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series Year II (XXXI)

without added
into
The blank sampleenzymes
muscle
tissue,
(M, control) - the
and the ageing
pieces of meatof the meat by
were
injectedthe action of
with 10% brine,exogenous and
without addingendogenous
enzymes;
enzymes
on
their
substrate.
Sample A the

brine used for theAfter the ageing,

injection
waseach sample was
supplied
withplaced in very
closed
enzyme.
Thewell
experimental
meat was injected
with 10% brinetubes and was
boiled in water
with
10
mg
bath
by
enzyme.
increasing
the
Sample
level
injected
was 15
meat;

B - thetemperature with
of
theabout 1oC/min.
enzymeuntil the sample
mg/100gachieved 70oC
on the thermal
this
Sample C - thecenter;
temperature was
level
of
the
maintained for
injected enzyme10
minutes.
was 20 mg/100gAfter boiling, the
meat.
samples
were
made
to
become
The brine used
for injection wascold
made up of: saltimmediately in
10g, sugar 10g,water bath with
tripolyphosphate ice; afterwards
they were kept in
2.5 g and water
refrigeration
86.5 g.
conditions for 12
The injection washours.
The
made manually,boiled samples
through
anwere brought at
injector with aroom
single needle, sotemperature,
that the entirewere taken out
quantity of brinecarefully from
could be pumpedthe experimental
in muscle mass.tubes, and after
The eliminatedremoving
the
brine was re-excess of juice
injected.
Thefrom the surface
meat injected was
using
filter
packed and stored
paper, they were
at the temperature
weighed.
The
of 4oC, for 24, 48juice
resulted
and 72 hours, toduring
the
allow
theboiling
was
diffusion of thecollected,
brine with or
weighed
and

used for different

analysis.
The
percentage
of
expressed juice
during thermal
treatment
was
calculated with
the relation:

% Expressed
juice = [Mass
of the meat
after ageing Mass of the
meat after
boiling and
cooling]
100/Mass of
meat after
ageing
2.5. Statistical
analysis
Five
experimental
batches
were
realized
for
each kind of
treatment.
Statistical
analysis, which
consist
in
evaluating the
mean
values,
standard error
and
standard
deviation with
the framing into
the confidence
interval of 95%,
was performed
using

Sigma
Plot
2001/Statistics
Data
software.
Experimental
data were fitted
using
Table
Curve
2D
software and the
regression
equations were
established based
on
statistical
criteria (r2, Fit
Standard Error or
F Statistic).

composition
of the beef
meat

higher levels of
papain
and
bromelin and
longer enzyme
Chemical components
action
times.
The
lowest
Moisture
level
of
Dry substance
hydrolysis was
Total nitrogen
noticed at the
Total Proteins
samples
that
Fats
were injected
Non-protein nitrogenwith
brine
Aminic nitrogen
without
Ammonia
enzymes,
in
which case the
The changes
3. Results and 3.2.
influence
of regarding the
Discussion
contents of non
exogenous
enzymes
on protein nitrogen
3.1. The
and free amino
composition of protein
acids
were
meat
hydrolysis
determined only
The experiments The proteolytic by
the
were made at the activity
of endogenous
laboratory level papain
and proteolytic
using the ham of bromelin on the enzymes of the
adult beef. In adult beef meat muscle
and
table
l,
are proteins
was those
reported
data estimated
by synthesized by
referring to the monitoring the the
chemical
variation of non psychrophilic
composition of protein
and microflora of
natural
adult beef meat. aminic nitrogen the
contamination
Analyzing these levels
during
of meat.
values it can be ageing.
seen that the
Both
papain The evolution
meat was well
and bromelin of non protein
chosen according
was
led to a limited nitrogen
to fats (5.52%
similar
to
all
hydrolysis
of
+1.24% fats) and
experimental
beef
meat
has
a
mean
proteins. The lots for the
content of water level
of entire process
of
75.8+1.3% hydrolysis was of ageing. The
and protein of more
levels of non
17.81+1.02%.
accentuated
protein nitrogen
Table 1.
when
using increased
Chemical

11

Ionescu et.al . / The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series, II (XXXI), 2008, 9-16

muscular rigidity,
than
myosin.
Myofibrillar
proteins are faster
hydrolyzed
by
papain when they
are
in
a
denaturated state
(Rattrie
and
Regenstein,
2000).
endopepti peptide
dases thatbonds
cleave thebetween
polypepti Arg, Lys,
des chainsand Gly
into
residues,
fragments but not on
with highthe bonds
molecular between
mass.
residues
Papain, of amino
Fi
compared acids with
g
to
acid
u
bromelin, character,
r
acted
which are
more
major in
e
intensivel the
y on themyosin
3
polypepti molecule
.
de
(42%).
T
fractions Papain
h
of
thehas
a
e
beef
different
muscle
i
and led to
higher
n
accumulat
f
ions
of
l
free
u
amino
e
acids.
n
Papain is
c
a cystein
e
proteinase
that
o
contains a
thiol
f
group Cys
p
27 in the
a
catalytic
p
site acting
a
like
a
i
nucleophi
n
lic group.
It
acts
a
upon the

continuously, varying action on the

in accordance with structural tissue
it
the applied treatment proteins:
hydrolyses
less
(Figure (1) and Figure
the actomyosin, a
(2)).
protein found in
muscular tissue
after
the
installation
of

Figure 1. The
influenc
e of
bromeli
n on
non
protein
nitrogen
accumu
lations

appreciate
the better
specificit
y
of
bromelin
for
the
proteins
of
beef
meat.
The
values
presented
in Figure
(3) show
that the
levels of
free
amino
acids, as a
main
compone
nt of non
protein
nitrogen,
increased
proportio
nally to
the
enzyme
levels.

Figure 2. The influence of papainFor both

on non proteinenzymes
nitrogen
the
accumulations increase

of
free
The highestamino
nitrogen
acid
accumulation levels was
s
wererelatively
recorded forsmall, due
tenderized to the fact
samples withthat the
used
bromelin,
and we canenzymes
are

d
br
o
m
el
in
co
nc
en
tr
at
io
n
o
n
th
e
fr
ee
a
m
in
o
ac
id
le
ve
ls

peptides
with
smaller
molecular
mass. The
increase
of
non
protein
fractions
improves
both the
beef meat
tenderizat
ion and
the
assimilati
on degree
of
nitrogeno
us
compoun
ds from
the beef
meat.

The
regression
curves
showed
that there
is a non
The aminiclinear
dependen
nitrogen
represents ce
only
onebetween
fraction
ofthe level
the
nonof
enzyme
protein
nitrogen, theadded and
rest
beingthe
composed ofaccumulat
12

ion of lowthe
molecular intensity
weight and
nitrogeno uniformit
y of the
us
compoun thermal
ds
astreatment,
result ofmeat
hydrolysi piece
s activitystructure
of papainand
thickness,
and
bromelin, which can
soluble incompete
the
trichloroa to
liberation
cetic acid
of soluble
(non
protein low
molecular
nitrogen,
weight
free
nitrogen
amino
compoun
acids)
ds. Values
Generally over 0.9
in
of
the
complex correlatio
system ofn
meat andcoefficien
particularl ts indicate
y in ourthat the
experime accumulat
ntal
ions
of
condition non
s, it is notprotein
excluded nitrogen
the
and free
participati amino
on
ofacids are
other
well
factors, correlated
such aswith the

enzymes
level that
was
added and
with the
time
of
action, in
case
of
both
bromelin
and
papain.
3.3. The
influence
of
exogenou
s
enzymes
on
the
soluble
proteins
The
influence
of
the
exogenou
s
proteolyti
c
enzymes
on
the
beef meat
was also
evaluated
by
determini
ng

The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series Year II (XXXI)

the solubility of
the protein in the
expressible juice
after the thermal
treatment.
The results of our Figure 4. The
study showed that
inf
proteolysis altered
lue
the properties of
nc
beef
meat
e
proteins,
by
of
increasing
its
the
solubility.
The
pa
losses of proteins
pai
in
expressible
n
juice
at
the
an
thermal treatment
d
were significantly
br
higher in case of
o
samples
treated
me
with
enzymes,
lin
compared
to
lev
control samples,
els
without enzymes
on
(Figure 4). The
the
increase
of
sol
enzyme level and
ub
of the enzyme
le
action time caused
pr
a
significant
ote
increase of the
in
protein losses in
los
the
expressible
ses
juice
at
the
boiling.
The
highest
breaking
ofThe
solubility
of
polypeptide
proteins,
in
the
chains,
the
case
of
the
weakening
of
treatment with
collagen network
both bromelin
and
thermal
and papain, was
denaturation of the
recorded for the
proteins, are, in
highest
tested
our opinion, the
level of enzymes
principal factors(20
mg/100g)
that contributed toand the longest
the elimination oftime of ageing
water and some(48 hours). The
soluble substancesolubility
of
from the meat.
enzymatically
treated
meat
proteins,
increased
differentially
after
boiling,

being 9 times
and 5.5 times
higher than the
one of the blank
sample, in case
of bromelin and
papain,
respectively. For
all studied cases
the
highest
correlation
coefficients
(r2>0.92) related
to
the
relationship
between
the
added enzyme
level and the
contents
of
soluble proteins
in
expressible
juice
after
boiling
was
obtained
by
modelling
the
experimental
data through a
non
linear
mathematic
equation.

3.4.
The
influence
of
exogenous
enzymes on the
protein
connective
tissue
Bromelin
and
papain
also
demonstrate
collagenase
activity
by
solubilisation of
collagen,
the
principal
constituent
of
connective tissue
which
enters
muscle
organization.
The
collagen
solubilisation
was

estimated througha compact gel.

the
These
hydroxyproline observations
accumulations thatsuggested the
varied with thefact that the
applied treatment,tenderization
by varying theeffect of the
bromelin or papainpapain may be
levels and themostly due to
tenderization time.the degradation
Compared to theof connective
tissue, which is
enzymatically
treated meat, theone of the four
level
ofprincipal factors
hydroxyproline involved in the
from the blanktenderization of
sample
wasmeat, together
with
the
significantly
shortness
of
the
lower, and was
the
due to the (i) freemuscle,
ageing
of
meat
hydroxyproline
normally presentunder the action
in
meat,
(ii)of endogenous
enzymes
and
hydroxyproline
the
marbling
obtained as a
degree of meat
consequence
of
(Miller et al.,
endogenous
1995).
collagenase action
and (iii) the one
taken out during
the
thermal
treatment applied
to the meat.

The action of
bromelin
was
more
intensive
upon
the
connective tissue,
compared
to
Figure 5. The
papain, leading to
a more advanced
solubilisation of
the
collagen
(Figure 5). It is
believed
that
papain affects the
mucoproteins and
the collagen of
the
connective
tissue of the
meat, more likely
than
other
structural
proteins. Under
papain action, the
collagen
suspensions are
transformed into

i
n
f
l
u
e
n
c
e
o
f
b
r
o
m
e

l
i
n
a
n
d
p
a
p
a
i
n
l
e
v
e
l
s
o
n
h
y
d
r
o
x
y
p
r
o
l
i
n
e
c
o
n
t
e
n
t
s

It is supposed
that proteases
act
on
the
proteoglycans
which link the
fibrillar
collagen II and
on the collagens
which act as

interfibrillar
connective
bridges.
tissue matrix, in
Nowadays
order
to
experimental
improve meat
studies
aretenderization
focused
on(Phillips et al.,
finding
2000).
innovative
The collagenase
methods
toactivity
of
disrupt
the

bromelin was
about
twice
higher
than
papain activity.
According
to
Ashie et al.
(2002),
the
bromelin
exhibits a more
13

Ionescu et.al . / The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series, II (XXXI), 2008, 9-16

accentuated
hydrolytic action on
collagen than on
myofibrillar proteins.
The effect of bromelin
or papain on the
connective
tissue,
which covers muscle
fibers
and
fiber
bundles, was also Figure
noticed by performing
a sensorial analysis on
the boiled meat. Our
results agree with the
observations of other
research groups who
tendered the beef meat
in marinade using level
of proteolytic enzymes
higher
than
0.01
UA/100 g meats.
3.5.
The
tenderization degree
and the losses at the
thermal treatment

6. The
variation
of
the
rigidity
Figure 8. The
index
variation
depending
of
the
on
the
rigidity
added
index
enzyme
dependin
level and
g on the
ageing
added
time
bromelin
(b
level and
oil
ageing
ed
time
me
at)

The influence of
bromelin and papain
on the tenderization
degree of the beef
meat was estimated
by establishing the
rigidity index, which
is a measure of the
resistance opposed by
the meat to the
compression test. This
parameter
was
Figure 7. The
evaluated for both
variation of the
raw meat and thermal
treated one and we rigidity index
could
note
a depending on
added
significant increase of the
papain
level
the rigidity index
and
ageing
value by increasing
(raw
the enzyme levels and time
meat)
their action time on
the substrate (Figure
6, 7 and 8), indicating
continuous
weakening of meat
structure during the
enzymatic ageing.

The
rigidity
index increased
exponentially
with the added
enzyme
levels
and
the
correlation
coefficients were
higher than 0.9,
for all studied
cases.
The
rigidity
index increased
exponentially
with the added
enzyme
levels
and
the
correlation
coefficients were
higher than 0.9,
for all studied
cases.

The enzymatic
ageing of beef

(ra
w
me
at)

meat with bromelin

or papain for 24 and
48 hours, led to the
improvement of the
hydrophilic
characteristics
of
meat proteins and to
the decrease of juice
losses at thermal
treatment compared
to
the
samples
without exogenous
enzymes (table 2).
By increasing the
levels of added
enzyme and of the
time of ageing, the
juice
losses
at

thermal
treatment
significantly
increased.
Table 2.
The losses
at thermal
treatment
Enzyme,
mg/100g
0
10
15
20
14

0
10
15
20

A
better
tenderization was
obtained
for
samples treated
with
bromelin
compared
to
those treated with
papain and to the
blank
sample.
Due
to
the
marked

Meat treated with papain

32.711.16
36.771.08
30.531.22
34.451.21
28.770.98
32.691.10
30.301.12
34.221.20

The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series Year II (XXXI)

hydrolytic
activity on the
structure
weakening of theconnective
tissue, leading
enzymatically
treated meat, oneto a better
should considertenderization
the
substantialof the adult
beef
meat.
decrease
of
From
the
boiling time in
analytical
order to avoid the
point of view,
pasty texture.
this fact was
counted
4. Conclusions through
determination
The developmentof the soluble
of adult beef meatcollagen from
texture
is
athe expressed
complex process,juice after the
which in ourthermal
experiment, wastreatment of
based on thethe
weakening
ofenzymatically
structural
treated meat.
elements of theIn order to
muscle, by usingavoid
the
exogenous
advanced
proteolytic
structural
enzymes such as
degradation of
papain
and
the meat, the
bromelin.
enzymatic
The main effecttreatment has
of papain andto be carefully
bromelin on themonitorized
beef
meatby limiting the
consisted
inlevels of the
limiting
papain
and
proteolysis of thebromelin and
proteins, whichthe
was emphasizedtenderization
through
theperiod.
dynamics of non
protein nitrogen,In conclusion,
we
free amino acidrecommend
and
solublethe following
protein levels ofparameters to
the juice resultedbe used for the
at the boiling ofenzymatic and
thermal
enzymatically
treated meat withtreatment of
different levels ofthe beef meat:
the
enzyme
enzymes and fordozedifferent periods10mg/100g
of time.
meat;
the
tenderization
Papain
and
time-24 hours,
bromelin showed

at
4oC,
followed by
a
thermal
treatment in
a
dynamic
regime
by
increasing
the
temperature
with 1oC/min
until
achieving
70oC in the
thermal
centre
to
completely
inactivate
papain and
bromelin.
References
Ashie, I.N.A.,
Sorensen,
T.L. and
Nielsen,
P.M.,
2002.
Effects of
Papain
and
a
Microbial
Enzyme
on Meat
Proteins
and Beef
Tendernes
s. Journal
of Food
Science,
67
(6),
2138
2142.
Bailey, A. J.,
1972. The
basis of
meat
texture.
Journal of
the
Science of
Food and
Agricultur
e,
23,
9951007.
Bailey, A. J.
And
Light, N.
D., 1989.

ConnectiveBertran,
J.A.,
Tissue in Meat Bonnet, M. adn
and
Meat Ouali, A., 1992.
Products. (ppComparative
action
on
1-355).
cathepsine
B
Elsevier
and
L
on
Applied
Science, Newintramuscular
collagen
as
York.
assessed
by
differential
scanning
calorimetry.
Meat Science,
32, 299-306.
Brooks,
J.
C.,
Belew, J. B.,
Griffin, D. B.,
Gwartney,
B.
L., Hale, D. S.,
Henning, W. R.,
Johnson, D. D.,
Morgan, J. B.,
Parish, F. C.,
Reagan, Jr., J.
O., Savell and J.
W.,
2000.
National beef
tenderness
survey1998. Journal
of Animal
Science, 78,
8521860.
Cheftel, J.C. and
Culioli,
J.,
1997. Effects of
high-pressure
on meat: A
review.
Meat
Science, 46 (3),
211-236.
Foegeding, E. A.
and Larick, D.
K.,
1986.
Tenderization of
beef
with
bacterial
collagenase.
Meat Science,
18, 201210.
Kang, C. K., Rice
and E. E., 1970.
Degradation of
various
meat
fractions
by
tenderizing
enzymes.
Journal of Food
Science,
35,
563577.

Koohmaraie, M.,
Babiker, A. S.,
Merkel, R. A.
and Dutson, T.
R., 1988. Role
of
Ca++dependent
proteases and
lysosomal
enzymes
in
postmortem
changes
in
bovine skeletal
muscle.
Journal
of
Food Science,
53,
1253
1257.
Koohmaraie,
M.,
1992a. Role of
neutral
proteinases in
postmortem
muscle
degradation
and
meat
tenderness. In
Proceedingd of
Reciprocal
Meat
Conference, 45

Koohmaraie, M.,
1992b.
The
role of Ca2+dependent
proteases
(calpains) in
post mortem
proteolysis
and
meat
tenderness.
Biochimie, 74,
239.
Koohmaraie, M.,
1994. Muscle
proteinases
and
meat
ageing. Meat
Science,
16,
93104.
Light,
N.,
Champion, A.
E., Voyle, C.
and Bailey A.
J., 1985. The
role
of
epimysial,
perimysial and
endomysial
collagen
in
determining
texture in six

bovine
muscles.and
Savell,
Meat Science, 13,J.W.,
1991.
137-149.
Using calcium
Miller, M.F., Hover, L.C.,chloride
to
Cook, K.D., Guerra,injection
improve
A.L.,
Huffman,
of
K.L., Tinney, K.S.,tenderness
beef
from
Ramsey,
C.B.,
Brittin, H.C. andmature cows.
Huffman,
L.M.,Journal of
1995.
ConsumerAnimal Science,
acceptability of beef69, 4469
4476.
steak tenderness in
T.,
the
home Nishimura,
and
restaurant. Journal Hattori, A. and
of Food Science, 60,Takahashi, K.,
1996a.
963-965.

Arrangement
Morgan, J.B., Miller,
and
R.K.,
Mendez,
identification of
F.M., Hale, D.S.

proteoglycans
in
basement
membrane and
intramuscular
connective
tissue
of
bovine
semitendinosu
s muscle. Acta
Anatomica,
155, 257-265.
Nishimura,
T.,
Hattori, A. and
Takahashi, K.,
1996b.
Relationship
between
degradation of
proteoglycans
15

Ionescu et.al . / The Annals of the University Dunarea de Jos of Galati

Fascicle VI Food Technology, New Series, II (XXXI), 2008, 9-16

connective tissue
and weakening of theproteins. Journal
intramuscular connectiveof Animal
tissue tissue during post-Science, 78,
mortem aging of beef.18611866.
Meat Science, 42, 251Rattrie, N.W. and
260.
Regenstein,
Penny, I. F., 1980. TheJ.M.,
2000.
enzymology
ofAction of crude
conditioning. In: Lawrie,papain on actin
R. A.: Developments in and
myosin
Meat Science-1 , (pp.heavy
chains
115-143).
Elsevierisolated
from
Applied
Sciencechicken breast
Publishers, Ltd., London. muscle. Journal
Stanton, C. and Light, N.,of Food Science,
1987. The effects of42 (5), 1159
conditioning on meat1163.
collagen:
Part Voisey,
1
P.W.:
Evidence for gross inInstrumental
situ proteolysis. Meat measurement for
Science, 21, 249265. food texture. In
Takahashi, K., 1996. StructuralDe Man, J.M.,
P.W.,
weakening of skeletalVoisey,
muscle tissue during post-Rasper V.F. and
D.W.,
mortem ageing of meat: Stanley
1976.
The
non-enzymatic
mechanism
of
meatRheological and
Texture in Food
tenderization.
Meat Science, 43, S67 Quality. (pp. 79141). CT, Avi
S80.
Publishing Co.,
Nishimura, T., Liu, A.,Westport.
Hattori,
A.
and
AOAC - Official
Takahashi, K., 1998.Methods
of
Changes in mechanical
Analysis.
15th
strength of intramuscular
ed. Association
connective tissue during
of
Official
postmortem aging ofAnalytical
beef. Journal of Animal Chemists,
Science, 76, 528 532. Washington, DC,
Phillips, A. L., Means, W. J., 1995
AOAC
- Official
Kalchayanand,
N.,
of
McCormick, R. J. andMethods
Miller, K. W., 2000.Analysis.
11th
Bovine
placentaled. Association
Official
protease
specificityof
Analytical
towards muscle
Chemists,

Washington, DC,
1984.
AOAC - Association
of
Official
Analytical
Chemists.
Methods
of
Analysis 15 th
Edition
Washington DC,
1990.
ISO

3496:1997.
Carne i produse
de
carne.
Determinarea
coninutului de
hidroxiprolin,
1999. pp A1.

1
6