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Extraction of bromelain from pineapple peels

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Food Science and Technology

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Extraction of bromelain from pineapple peels

S. Ketnawa, P. Chaiwut and S. Rawdkuen
Food Science and Technology International 2011 17: 395 originally published online 3 August 2011
DOI: 10.1177/1082013210387817
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Article

Extraction of bromelain from pineapple peels

S. Ketnawa1, P. Chaiwut2 and S. Rawdkuen1

Abstract
Large amount of pineapple peels (by-products) is left over after processing and they are a potential source for
bromelain extraction. Distilled water (DI), DI containing cysteine and ethylenediaminetetraacetic acid (EDTA)
(DI-CE), sodium phosphate buffer pH 7.0 (PB) and PB containing cysteine and EDTA (PB-CE) were used as
extractants for bromelain from the pineapple peels. The highest bromelain activity was obtained when it was
extracted with PB-CE (867 and 1032 units for Nang Lae and Phu Lae cultv, respectively). The PB could
maintain the pH of the extract (pH 5.15.7) when compared with others. Under sodium dodecyl sulfate
polyacrylamide gel electrophoresis, the extract showed protein bands in the range 2428 kDa. The protein
band with a molecular weight of 28 kDa exposed the clear zone on blue background under the caseinsubstrate gel electrophoresis. The effects of the bromelain extract on the protein patterns of beef, chicken and
squid muscles were also determined. Trichloroacetic acid soluble peptide content of all the treated muscles
increased when the amount of bromelain extract increased. Decrease in myosin heavy chains and actin was
observed in all the muscle types when bromelain extract was used. The best extractant for bromelain from
pineapple peels was PB-CE. Moreover, bromelain extract could be used as a muscle food tenderizing agent
in food industries.

Keywords
Extractant, bromelain, pineapple peel, TCA-soluble peptides, meat tenderization
Date received: 3 June 2010; revised: 17 August 2010

INTRODUCTION
Bromelain, the proteolytic enzyme, is found in the tissues of the plant family Bromeliaceae of which pineapple (Ananas comosus L. Merryl) is a well-known source.
It is also found in pineapple wastes such as cores, peels
and leaves in relatively smaller quantities as compared
to stems and fruits (Maurer, 2001; Sriwatanapongse
et al., 2000; Umesh et al., 2008). The stem bromelain
(EC 3.4.22.32) and fruit bromelain (EC 3.4.22.33)
obtained from the pineapple stem and pulp are commercially available. Pineapple wastes, especially peels
and cores, are left in large amounts (4050%) after
fresh-cut processing in Chiang Rai, Thailand
(24 tons per year; DOAE, 2009). Normally, these
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wastes were used as feed for cattle or fertilizer.

However, it is still very low in value when compared
to the value of the substantial amount of bioactive compounds contained in those waste materials, especially in
bromelain or vitamin C (Umesh et al., 2008). Because
pineapple peels make up such a large amount of postprocessing waste material (3040%; Ketnawa et al.,
2010), they seem to be an available source for signicant bromelain extraction.
In order to isolate intracellular protein/enzymes
from any source, cells must be disrupted to release
the target proteins in a soluble from. The success of
1
Food Technology Program, School of Agro-Industry, Mae Fah
Luang University, Muang, Chiang Rai 57100, Thailand.
2
School of Cosmetic Science, Mae Fah Luang University, Muang,
Chiang Rai 57100, Thailand.

Corresponding author:
S. Rawdkuen, Food Technology Program, School of AgroIndustry, Mae Fah Luang University, Muang, Chiang Rai 57100,
Thailand
Email: saroat@mfu.ac.th

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Food Science and Technology International 17(4)

cell disruption depends on the number of variables
such as the choice of buers, the presence of protease
inhibitors and the osmolarity of the re-suspension
buer (Ahmed, 2004). The condition and the constituent of the extraction buer depend on the nature of the
cell type, the target protein and its intended application.
Proteins/enzymes are extremely heterogeneous biological macromolecules. Their properties can be severely
aected by small changes in the extraction buering
system such as in hydrogen ion concentration, ionic
strength and acidbase. Chelating agents such as ethylenediaminetetraacetic acid (EDTA) and ethylene glycol
tetraacetic acid (EGTA) are normally contained in the
buer system, because they protect enzymes from
inactivation by heavy metals and proteolysis by metalloprotease (Ahmed, 2004).
Bromelain has been used widely in food, medical,
pharmaceutical and cosmetic industries and other
industries as well. In the food industry, it is used for
meat tenderization, grain protein solubilization, beer
clarication, baking cookies and protein hydrolysate
production (Walsh, 2002). For medical and pharmaceutical industries, bromelain is well known for clinical
and therapeutic applications, particularly for modulation of tumor growth, third-degree burns, improvement
of antibiotic action, and as a drug for the oral systemic
treatment of inammatory, blood coagulation related
and malignant diseases and digestion (Maurer, 2001).
In cosmetics, bromelain is used as an active ingredient
in western face and body care products to provide
gentle peeling eects (removing stratum corneum
cells; Aehle, 2007). In addition, bromelain is also used
for skin pre-tanning, softening and bating in leather
industries (Walsh, 2002), improving the dyeing properties of protein bers (Koh et al., 2006) and decomposing or partially solubilizing protein bers from silk and
wool (Koh et al., 2006; Singh, 2003).
Since bromelain oers multiple benets in various
industries, the extraction and characterization of the
enzyme from pineapple wastes are interesting for
its rich properties and also for waste utilization.
Therefore, the objective of this study was to investigate
the extraction of bromelain from pineapple peels using
dierent extractants. Characteristics and eects on
muscle food samples were also determined.

MATERIALS AND METHODS

Materials
Chemicals and raw materials. Sodium di-hydrogen
orthophosphate (NaH2PO4.2H2O) and di-sodium
hydrogen phosphate (Na2HPO4) were purchased from
Ajax nechem Chemical (Seven Hills, NSW, Australia).
Cysteine, tyrosine, EDTA, casein and bovine
serum albumin (BSA) were obtained from Fluka

(Buchs, Switzerland). Trichloroacetic acid (TCA) was

procured from Merck (Darmstadt, Germany). Betamercaptoethanol (bME), Coomassie Brilliant Blue
G-250, sodium dodecyl sulfate (SDS) and stem bromelain (EC. No. 253-387-5) were purchased from Sigma
Chemical Co. (St. Louis, MO, USA).
The pineapple peels (Nang Lae (NL) and Phu Lae
(PL) cultv.) were obtained from a shop in Nang Lae
subdistrict, Muang, Chiang Rai, Thailand. The peel
was washed, air-dried and then stored at 4  C for further experiments.
Methods
Crude bromelain extract preparation. Four extractants were used in this experiment: distilled water
(DI), distilled water with 15 mM cysteine and 2 mM
EDTA (DI-CE), 100 mM sodium phosphate buer
pH 7.0 (PB) and 100 mM sodium phosphate buer
pH 7.0 with 15 mM cysteine and 2 mM EDTA (PBCE). The pineapple peel was blended in cold extractants at a 1:1 ratio (w/v) for 3 min and then ltered
through a cheese cloth. The ltrate was centrifuged at
10 000 g for 20 min at 4  C. The obtained supernatant
was referred to as crude bromelain extract: BE and
was used for the experiments.
Characterization of bromelain
Determination of protease activity. The protease
activity of the bromelain extract was determined
according to the method in Murachi (1976) using tyrosine as a standard. The bromelain activity was determined using casein (1.5%, w/v) as a substrate in the
presence of cysteine and EDTA at 37  C and pH 7.0
for 10 min. After exactly 10 min, the reaction was
stopped by adding 3.0 mL of 5% (w/v) TCA.
Precipitated protein was removed by centrifugation at
10 000 rpm for 10 min. The absorbance of the clear
supernatant was measured at 275 nm. One unit of protease activity is dened as the amount of enzymereleasing product equivalent to 1 mmol of tyrosine
min/mL under the assay conditions.
Protein determination. The protein concentration
of the samples was determined using the Bradford
(1976) method and BSA was used as a standard.
Protein patterns. The molecular weight (MW) distribution was determined using SDSPAGE (PAGE,
polyacrylamide gel electrophoresis) according to the
Laemmli (1970) method. The sample was mixed with
the sample buer (0.5 M TrisHCl, pH 6.8, containing
4% (w/v) SDS and 20% (v/v) glycerol) with and
without bME. The proteins were loaded into the
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Ketnawa et al.
polyacrylamide gel (15% running and 4% stacking
gels) and subjected to electrophoresis at a constant
current of 15 mA/gel. After separation was achieved,
the protein was stained with 0.02% Coomassie
Brilliant Blue R-250 and destained with a mixture of
aceticmethanol solution.
Activity staining. The separated protein by electrophoresis was veried for bromelain by using activity
staining (casein-substrate gel electrophoresis) as
described by Garcia-Carreno et al. (1993). The gel
was immersed in 50 mL of 2% (w/v) casein 50 mM of
PB, pH 7.0 for 45 min with constant agitation at 4  C.
The gel was then incubated at 37  C for 30 min with
constant agitation. The development of a clear zone
on the blue background indicated bromelain activity.
Effect of crude bromelain
proteins degradation

extract

on

muscle

Marination of meat samples. Two grams of meat

samples (beef, chicken and squid) were marinated with
BE (1000 and 2000 mL, the activity of 6.36 and
12.72 units, respectively) at room temperature for 1 h.
The control samples were marinated using DI instead
of BE. The marinated samples were used to determine
the eect of BE on protein degradation using TCAsoluble peptide content and hydrolysis patterns by
SDSPAGE.
Determination of TCA-soluble peptide content.
TCA-soluble peptide content of treated samples incubated at room temperature for 1 h were measured by

the method used in Rawdkuen and Benjakul (2008) The

treated samples (2 g each) were homogenized in 18 mL
of 5% (w/v) TCA for 1 min. The homogenate was kept
at 4  C for 1 h and centrifuged at 8000  g for 5 min.
The soluble peptides in the supernatant were measured
according to the Lowry method (Lowry et al., 1951).
The TCA-soluble peptide content was calculated as millimoles of tyrosine per gram of sample.
Degradation of muscle proteins. The marinated samples (2 g) were solubilized with 18 mL of 5% (w/v) SDS
and then heated at 85  C for 1 h before being subjected
to electrophoresis as previously described.
Statistical analysis. An analysis of variance was used
to analyze the data from triplicate measurements.
Dierences between means were evaluated by
Duncans multiple range test using the SPSS statistic
program (version 11.5).

RESULTS AND DISCUSSION

Bromelain extraction from pineapple peels
The pH, protein content and protease activity of bromelain extract from pineapple peels are presented in
Table 1. The volume of BE ranged from 143 to
149 mL for NL. The highest volume amount was
observed when it was extracted by DI-CE
(148.67 mL). The volume of BE of PL ranged from
152 to 162 mL from 100 g of the peel. Both NL and
PL showed higher extraction volumes when using the
extractant containing cysteine and EDTA when compared to that without these compounds. For the pH

Table 1. Characteristics of crude bromelain extracts from 100g of pineapple Nang Lae (NL) and Phu Lae (PL) peels using
different extractants

Extractant Volume (mL)

NL peel
DI
DI-CE
PB
PB-CE
PL peel
DI
DI-CE
PB
PB-CE

143.67  1.00
148.67  0.50
143.67  2.78
146.00  0.87

Protein
content
(mg/mL)

pH

Total
protein (mg)

Activity
(unit/mL)

Specific
activity
(unit/mg
protein)

Total activity
(unit)

a
c
a
b

3.64  0.18
3.58  0.12
5.15  0.01
5.14  0.02

b
a
c
d

0.337  0.02
0.335  0.01
0.702  0.01
0.564  0.02

a
a
c
b

48.48  2.85
49.81  2.19
100.79  1.18
82.41  3.55

a
a
c
b

2.28  0.02
2.16  0.04
2.76  0.04
5.74  0.06

b
a
c
d

327.71  3.51
321.01  6.07
396.44  7.53
866.88  3.73

b 6.77  0.35
a
6.45  0.34
c
3.93  0.05
d 10.54  0.47

155.67  1.73 c
152.00  1.73 a
153.00  2.29 b
161.6  2.50 d

3.75  0.02
3.75  0.03
5.83  0.03
5.74  0.03

a
a
c
b

0.215  0.01
0.257  0.01
0.574  0.02
0.625  0.02

a
b
c
d

33.47  1.46
40.89  1.86
87.79  2.89
101.00  4.56

a
b
c
d

2.85  0.08
3.15  0.12
5.97  0.19
6.98  0.11

a
b
c
d

443.66  16.47
478.90  13.68
913.77  20.84
1031.94  21.51

a
b
c
d

b
b
a
c

13.26 0.34 d
11.79  0.89 c
10.41  0.27 a
10.23  0.26 b

Means  SD (n 3). Different letters in the same column indicate the significant differences (p < 0.05).
DI, distilled water; DI-CE, distilled water with 15 mM cysteine and 2 mM EDTA; PB, 100 mM phosphate buffer pH 7.0; and PB-CE, 100 mM
phosphate buffer pH 7.0 with 15 mM cysteine and 2 mM EDTA.

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Food Science and Technology International 17(4)

values of BE in both cultivars, PB and PB-CE seemed
to retain the most pH of the crude extract at around
5.1 in NL and 5.7 in PL, while DI and DI-CE exhibited
pH of 3.6 and 3.7 for NL and PL, respectively. The PB
has a buering capacity to retain the system pH solution by slightly changing the pH value (Bonner, 2007).
Therefore, it is benecial to extract enzymes by promoting the maximum activity. The use of buer solution is
sometimes very ecient and is acceptable for some proteins/enzymes without causing excessive denaturation
(Bonner, 2007). The best extractants used in the extraction process of any enzymes should maintain the system
pH close to the optimum pH of the target enzyme.
It should not alter the enzyme activity. As already
known, bromelain has an optimum pH at around
4.04.5 and 6.8 for stem and fruit bromelain, respectively (Harrach et al., 1995; Rowan et al., 1990) and the
extractant buer that can keeps the system pH close to
that value should be selected.
Extracting bromelain using PB allowed for higher
protein content and protease activity compared to
that using DI. High protein content and protease activity could be attributed to the eect of salts present in
the buer that could solubilize the protein out of the
compact structure in the sample. BE from NL showed
the highest protein content (100.79 mg) when PB was
used, while PB-CE provided the highest protease activity (866.88 units). The highest specic activity of bromelain was found in the extract using PB-CE (10.54)
and DI (13.26) for NL and PL, respectively. However,
the protease activity obtained from the DI of PL was
still 2.3 times lower than that of PB-CE. The use of
exogenous proteases with a view to apply their proteolytic potentials to tenderize muscle ranges from 6 to
600 units per 100 g of sample. However, the concentration used depends on the purity of the enzyme as well as
the type of muscle. High amount of exogenous proteases is needed for food application compared with
pharmaceutical or cosmetic products. In addition, the
purity of the enzyme used for medicinal products concerned more about the purity and specic function.
From the result obtained, high activity of crude
enzyme extract would be useful in food application
rather than in cosmetic or pharmaceutical products.
The dierent protein and enzyme recovery when
using dierent extractants was probably due to the ability and quantity of active compounds contained in the
samples. As already known, most proteases in pineapple
belong to cysteine proteases and contain cysteine residue
in their active sites; so, when the extractant is contained
with their activator, a higher enzyme recovery was
obtained. This might be because proteolytic activities
are activated by additions of small reducing agents
such as cysteine and chelating agents like EDTA
(Chaiwut et al., 2007). In the presence of cysteine, it

prevents the disulde bonds of the protein/enzyme cysteine molecules from reducing (Sluyterman, 1967). EDTA,
can trap heavy metals that enhance the oxidation of
thiols with molecular oxygen and they can form complexes with specic groups, which may cause problems
like lowering enzyme activity (Janson and Ryden, 1998).
Between NL and PL peel extracts, the latter provided
higher total activity than the former. This is because
PL peels gave a high volume of the extract when using
the same amount of the starting material. According to
the results, the highest total activity of the extract was
found when PB-CE was used for both NL and PL peels.
Characteristics of bromelain from
pineapple peels
Protein patterns. The protein patterns of BE from NL
and PL with dierent extractants are shown in Figure 1.
In the reducing condition, the migrations of proteins in
BE were quite similar in all the extractants used. BE
from both cultivars showed the protein composition
MW to be in the range 2428 kDa. The main protein
component found in BE of NL and PL has an MW of
around 28 kDa. However, the smear protein bands with
the MW lower than 18.3 kDa were also observed in all
samples. The reducing agent (bME) could split the protein molecule which stabilized with a disulde bond
into a dierent low MW protein (Walsh, 2002). The
commercial stem bromelain (lane ST) exhibited the protein band at an MW of 28 kDa. For the non-reducing
condition, the migrations of proteins in BE were quite
the same as that of the reducing condition. Dierent
protein patterns using DI (numbers 1 and 2) and PB
(numbers 3 and 4) were observed in the non-reducing
condition. The two major protein bands in the nonreducing condition of the samples extracted with DI
(numbers 1 and 2) were found at around 28 kDa and
below 18.3 kDa, while only a single protein band
(MW  28 kDa) was observed when using PB
(number 3 and 4) as an extractant. Nonetheless, the
BE of NL extracted by PB and PB-CE showed other
major protein bands at around 30 and also below
18.3 kDa in some samples. Dierences in protein components among those BEs are probably due to the differences in the amount and characteristics of interfering
proteins obtained from using dierent extractants.
Activity staining. To verify the band of bromelain,
activity staining was performed (data not showed).
No clear zones were observed for the protein band in the
reducing condition. The presence of a reducing agent,
such as bME or dithiothreitol, breaks the disulde
bonds in the protein structure, making the protein less
conformationally stable and forcing it to lose functional and/or structurally important elements of the
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Ketnawa et al.

Reducing

Non-reducing

Figure 1. SDSPAGE patterns of crude bromelain extract from Nang Lae (NL) and Phu Lae (PL) pineapple peels
extracted by (1) DI; (2) DI-CE; (3) PB; and (4) PB-CE in reducing and non-reducing conditions.
Protein (6 mg) was loaded into the gel for protein staining. ST: stem bromelain. M: MW markers.

domain tertiary structure (Ahmed, 2004; Walsh, 2002).

This reason can be used to explain why there was no
apparent clear zone in the substrate gel of the reducing
condition. The clear zone generated by the proteolytic
activity on the blue background was present in the protein band at around 28 kDa. Clear zones were observed
in all BEs and in the commercial stem bromelain, but
not in the BE of NL using PB as an extractant. This is
probably due to a rather low bromelain content and
purity. Comparing the pineapple peels with the stems,
the MW of bromelain extracted from the peels gave a
slightly lower MW than that from the stem bromelain.
Pineapple extract also contains peroxidase, acid phosphatase, several protease inhibitors and organically
bound calcium, which may play a role in bromelain
inhibition or in the cleavage protein structure, resulting
in less activity (Bitange et al., 2008; Umesh et al., 2008).
It is noticed that the major protein band with an MW
of around 28 kDa exhibited caseinolytic activity. This
implies that the pineapple peel possibly contains other
proteases apart from the bromelain, which is showing
activity at an MW of around 28 kDa.
Effect of BE on muscle proteins
TCA-soluble peptide content. Eect of the addition
of bromelain extracts to protein degradation was investigated in beef, chicken and squid muscles. The TCAsoluble peptide content of the treated samples are presented in Table 2. The highest TCA-soluble peptide

content in all the muscle samples was found in the

BE-treated samples extracted using PB-CE followed by
DI-CE, PB and DI, respectively. When the amount of BE
increased, the TCA-soluble peptide content increased in
every muscle sample. TCA-soluble peptide content in the
squid muscle was the highest when compared with other
muscle foods. High values of TCA-soluble peptides in
squid may be caused by the synergistic eects of endogenous proteases presented in this muscle (Cortes-Ruiz
et al., 2008). Ramirez-Olivas et al. (2004) have suggested
that jumbo squid mantle muscle possesses high proteolytic activity. Degradation of the myobrillar proteins
resulted in an increase in peptides and free amino acids
(Naveena et al., 2004). The tyrosine level indicated that
the endogenous oligopeptides and/or free amino acids, as
well as degradation products, accumulated after being
marinated with BE. From the result, high TCA-soluble
peptide content indicated greater hydrolysis of the muscle
proteins generated by proteolytic enzymes in BE. In addition, high hydrolytic activity was found in the samples
treated with BE using extractants containing an enzyme
activator.
Protein patterns of muscle samples. The hydrolytic
patterns of muscle proteins treated with BE are
shown in Figures 2 and 3 for BE from NL and PL,
respectively. The two main protein components in the
beef and chicken muscle were myosin heavy chain
(MHC) and actin (AC), while only AC was found as
the major one in the squid muscle. The decrease in

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Food Science and Technology International 17(4)

Table 2. TCA-soluble peptide contents in muscle samples treated with crude bromelain extract from Nang Lae (NL) and
Phu Lae (PL) pineapple peels
TCA-soluble peptide (millimoles of tyrosine per gram of sample)
Pineapple

Muscle

BE (mL)

DI

NL

Beef

1000
2000
1000
2000
1000
2000

1.11  0.01
1.50  0.02
1.27  0.02
1.52  0.01
1.64  0.01
1.97  0.01

b, A
b. A
b. B
c, A
a, C
a, B

1.37  0.01
1.79  0.01
1.38  0.01
1.60  0.01
2.04  0.00
2.21  0.01

c, A
c, B
c, A
d, A
b, B
b, C

1.09  0.01
1.32  0.01
1.10  0.01
1.32  0.01
2.55  0.01
2.86  0.01

a,
a,
a,
a,
c,
c,

A
A
B
A
C
B

1.86  0.01
2.02  0.01
1.37  0.01
1.61  0.01
2.68  0.01
2.77  0.07

d, B
d, B
c, A
d, A
d, C
d, C

1000
2000
1000
2000
1000
2000

0.97  0.00
1.30  0.00
1.12  0.01
1.32  0.01
1.88  0.00
1.92  0.01

a,
a,
a,
a,
a,
a,

1.21  0.01
1.58  0.02
1.32  0.01
1.65  0.15
2.03  0.02
2.39  0.01

b, A
b, A
c, B
c, B
b, C
b, C

1.76  0.01
2.07  0.01
1.55  0.01
1.77  0.02
2.42  0.11
2.50  0.01

d, B
d, B
d, A
d, A
c, C
c, C

1.55  0.01
1.78  0.01
1.17  0.01
1.59  0.01
2.45  0.01
2.87  0.01

c, B
c, B
b, A
b, A
d, C
d, C

Chicken
Squid

PL

Beef
Chicken
Squid

DI-CE

A
A
B
B
C
C

PB

PB-CE

Means  SD from triplicate determinations. Mean values followed by different small letters in the same row indicate significant differences
(p < 0.05). Mean values followed by capital letters in the same column indicate significant differences (p < 0.05).

DI

PB

DI-CE

PB-CE

Figure 2. SDSPAGE patterns of muscles treated with crude bromelain extract from NL pineapple peels extracted by
different extractants.
BC, CC and SC, controls without extract; B, beef; C, chicken; and S, squid. The numbers 1 and 2 indicate the amount of
extract, 1 and 2 mL, respectively. MHC, myosin heavy chain; AC, actin; TM, tropomyosin; TN-T, troponin-T; and MLC,
myosin light chain.

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Ketnawa et al.

DI

DI-CE

PB-CE

PB

Figure 3. SDSPAGE patterns of muscles treated with crude bromelain extract from PL pineapple peels extracted by
different extractants.
BC, CC, and SC, control without extract; B, beef; C, chicken; and S, squid. The numbers 1 and 2 indicate the amount of
extract, 1 and 2 mL, respectively. MHC, myosin heavy chain; AC, actin; TM, tropomyosin; TN-T, troponin-T; and MLC,
myosin light chain.

MHC and AC was observed in all the meat samples

when the amount of BE added increased (numbers 1
and 2). Small peptides or protein bands with an MW
lower than the myosin light chain were clearly observed
when BE was applied. Compared to the BE of NL
(Figure 2) and PL (Figure 3), the highest hydrolytic
activity was found in the samples treated with BE
from PL (Figure 3) as indicated by the low band intensity of the major protein components. There was
increased muscle protein proteolysis in all enzyme-treated samples as evidenced by the reduction in number
and intensity of protein bands when the BE concentration was increased. Squid muscle was more hydrolyzed
when compared with others. This result conformed to
the TCA-soluble peptide content as previously mentioned. In addition, beef muscle was more susceptible
to BE than that of the chicken muscle as indicated by
the low number and intensity of the protein bands,
especially in the degradation bands of proteins. From
the results, it is also evident that cleavage of high MW

proteins into lower ones was generated. In addition,

large amounts of high protein breakdowns were
observed when the sample was treated with BE containing an enzyme activator. Jorgova et al. (1989) reported
that the reduction of high MW fractions in muscle proteins treated with proteolytic enzymes was related to
increasing meat tenderness. Proteolytic degradation
of myobrillar proteins, especially myosin, resulted in
a reduction in MW and the loss of their functional
properties (Visessanguan et al., 2003).

CONCLUSION
The extractants played an important role in maintaining bromelain activity from the pineapple peel. PB-CE
showed to be the most ecient in bromelain extraction
provided by the BE with the highest activity for both
cultivars. TCA-soluble peptide contents and hydrolytic
patterns of muscle proteins conrmed that the extractant containing cysteine and EDTA could better

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Food Science and Technology International 17(4)

maintain the enzyme activity. As a consequence, the
most appropriate extraction buer for bromelain
extraction from pineapple peels would be PB-CE.
ACKNOWLEDGMENTS
The authors express their sincere thanks to Mae Fah Luang
University for partially nancing this study. They also thank
Prof. Matthew Robert Ferguson, Language Center, Naresuan
University for kindly providing suggestions and corrections
for the manuscript.

FUNDING
This work was supported by Mae Fah Luang University and
the National Research Council of Thailand [grant number:
PK/2553-40].

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