KHO ST KH NNG LM MM THT CA ENZYME BROMELIN

THU C T PHN PH LIU DA - CHI NGN

L Th Thanh Mai1, Nguyn Kiu Hng2

Da l loi thc vt rt c bit, va l mt loi cy n tri, va l ngun

nguyn liu to ra nhng sn phm c gi tr thng phm cao nh: enzyme
bromelin dng trong thc phm, y hc. Vit Nam c nhiu nghin cu
nhm tm ra quy trnh thu nhn enzyme bromelin t cc phn khc nhau ca
cy da c bit l t ph liu ca cc nh my da ng hp, nhng vic
dng enzyme bromelin lnh vc ch bin thc phm-lm mm tht cha c
quan tm. Trong bi bo ny chng ti trnh by kt qu nghin cu xc nh
iu kin ti u cho qu trnh lm mm tht ca enzyme bromelin - sn phm
thu c t phn ph liu da - chi ngn (hnh s 1), lm c s sn xut bt
lm mm tht.

Hnh s 1.Chi da sau khi lm sch l chi

I. NGUYN LIU V PHNG PHP

Ph liu da-phn chi ngn (Cng ty Thc phm Xut khu Tn Bnh).

Chun b enzyme bromelin t phn ph liu chi ngn da (Bro -C) theo
quy trnh tm tt c trnh by S s 1.

Enzyme bromelin ca hng Merck (Bro -M)

Tin s, Trng Khoa Cng ngh Sinh hc, H M TP.HCM

Cng ty Thanh Sn Ha nng

-1-

Tht b dai.

Choi

Lam sach la

Ra

Nghien, ep

Ba

Dch ep
Ly tam
Con (4:1)
Lu : Sau khi lm sch l, chi

Dch chiet
e 5oC, 2 gi

khng b chuyn sang mu nu

en; Cn c lm lnh trc
khi lm tc nhn gy ta; Dch
p chi khng b en.

Ly tam lanh
Tua t
Say kho, 45oC
Tua kho
(Bro-choi)

S s 1: S thu nhn bromelin t 5-10 kg chi lm sch

l

Xc nh hot tnh protease theo phng php Dole(1) (EDU) vi c cht l

casein, dung dch m phosphat 0,05M, pH 7. Nng casein trong hn
hp phn ng l 0,6%. Tng th tch hn hp phn ng l 11 ml. Hot
enzyme c xc nh 37oC trong 10 pht. Lm ngng phn ng bng
dung dch TCA 0,11M trong sodium acetat 0,22M v acetic acid 0,33M.
Mt n v EDU l lng enzyme lm tng s hp thu ca 1 g Tyrosine

trong 1 pht.

Xc nh hot tnh ng t sa (MCU)(2)

nh gi mc tc ng ca ch phm enzyme bromelin, ngi ta

thng th hin qua cc n v hot tnh nh GDU (gelatin dissolving units),

MCU (milk clotting units), FIP (Federation Internationale du Pharmaceutiques)

-2-

hoc nhng n v do chnh cc nh sn xut t ra cho sn phm ca h nh

Rorer units (Rhne Poulenc Rorer Pharmaceuticals, Inc.). Phn ln cc n v
ny thng khng th so snh hoc quy i c. Hin nay, nh gi
mnh ca enzyme bromelin, ngi ta thng s dng cc n v nh MCU,
GDU.

Cc phng php ho sinh thng dng trong phng th nghim.

Phng php xc nh nng enzyme thch hp lm mm tht:

Chn ty nng enzyme t thp n cao. Cn trng lng ca mu

tht v chn nng enzyme xc nh tnh ra trng lng ca enzyme tng

ng vi nng trn. Cho ht lng enzyme ny ha tan trong 20ml nc ct.
Ngm tng mu tht trong tng nng enzyme khc nhau trong thi gian 5
pht. Ly mu tht ra em ch bin trong 2 pht, th cm quan, chn nng
enzyme bromelin thch hp lm mm tht.

Xc nh thi gian ti u enzyme bromelin lm mm tht

Cn tht, ngm mu tht trong nng enzyme thch hp trong cc thi

gian khc nhau: 3, 5, 7, 9, 11 pht. Ly tht ra, em ch bin trong vng 2

pht, th cm quan xc nh thi gian thch hp tht mm mc va
phi.
II. KT QU V THO LUN
1. iu kin ti u lm mm tht b bng Bro-M
a. Nng Bro-M
Th nghim c tin hnh ln lt trn cc mu tht p Bro-M
(384,6 MCU/mg enzyme) cc nng khc nhau trong cng thi gian 5 pht.
Kt qu thc nghim c trnh by Bng s 1.

Bang so 1.nh hng ca nng Bro-M n mm ca tht

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Mu

Khi
lng tht

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

(g)
16,97
14,09
12,58
9,94
16,21
11,51
12,09
12,43
12,53
11,34
11,16
10,42
12,65
12,13
10,97
11,25
11,72
12,12
12,50
11,59
11,28
11,38

Enzym
Khi lng
% theo
(mg)

lng tht

8,48
14,5
18,9
19,9
40,5
34,5
42,3
49,7
56,4
56,7
61,4
62,5
82,2
84,9
82,3
90,0
99,5
109,0
118,8
115,9

0,05
0,10
0,15
0,20
0,25
0,30
0,35
0,40
0,45
0,50
0,55
0,60
0,65
0,70
0,75
0,80
0,85
0,90
0,95
1,00

124,1
136,5

1,10
1,20

Thi gian

Cm quan

thy phn
(pht)
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5

Tht rt dai
Tht rt dai
Tht rt dai
Dai
Dai
Dai
Dai
Dai
Dai
Dai
Dai
Dai
Hi dai
Hi dai
Hi dai
Hi mm
Hi mm
Hi mm
Mm, n hi tt
Mm, n hi hi

km
Hi r, v hi

ng
Hi r, c v
ng

Kt qu cho thy, nu dng nng enzyme thp (0,9% so vi khi

lng tht), tht vn cn cng. Ngc li, nu nng enzyme cao (1,1% so
vi khi lng tht), tht b r, v c v ng. Nng 0,95% (tng hot tnh l
365,37 MCU) l tt nht lm mm tht.
b. Thi gian
Th nghim c tin hnh ln lt trn cc mu tht p Bro-M
(0,95%) trong cc thi gian khc nhau. Kt qu c trnh by Bng s 2.

Bang so 2.nh hng ca thi gian p n mm ca tht (0,95%

enzyme bromelin/khi lng tht)

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Thi gian

Khi

Khi lng

p (pht)

lng tht

enzyme

(g)
12,41

(mg)
117,8

Tht hi cng, c n hi tt, tht c

12,44

118,2

v ngt
Tht mm, n hi tt, b mt tht

121,1

mn, tht c v ngt

Tht mm, n hi km, v chua

119,8

ng
Tht hi r, khng cn n hi, c

121,9

v ng
Tht r, c v ng

12,75

12,62

11

12,84

Cm quan

Thi gian thy phn tt nht l 5 pht. Nu thi gian ngn hn th khng
lm tht mm, cn nu lu qu th tht c v ng.
Nh vy, khi dng enzyme lm mm tht b dai th nng ca
enzyme v thi gian p enzyme nh hng nhiu n mm ca tht. Nu
nng enzyme cng cao, thi gian p enzyme cng lu tht cng mau mm.
Nng enzyme qu cao s lm tht c v ng nn nng enzyme thch hp
lm mm tht l 0,95% v thi gian p thch hp l 5 pht. Ngc li nng
enzyme v thi gian p enzyme cng ngn th tht lu mm hn.
c. Hm lng m amin, m amoniac c hnh thnh
trong cc mu th nghim
Mi mu c lm th nghim 10 ln ly gi tr trung bnh cng. Kt
qu thc nghim c trnh by Bng s 3.

Bang so 3.Hm lng nit formol

Mu
Khi lng tht (g)
Khi lng enzyme (mg)
Th tch NaOH 0,1N (ml)
Lng nit formol/mu tht (g)
Lng nit formol/kg tht (g)

1
12,490
0
2,05
32,5.10-4
0,229

Ghi ch:

Mu 1: mu tht c r ng
-5-

2
12,339
117,05
2,85
43,8.10-4
0,321

3
12,412
118,52
3,20
49,1.10-4
0,355

4
12,278
117,66
3,50
54,8.10-4
0,400

 Mu 2: mu tht c p enzyme trong thi gian 3 pht

Mu 3: mu tht c p enzyme trong thi gian 5 pht
Mu 4: mu tht c p enzyme trong thi gian 7 pht
Mu 1 c lng nit formol thp nht, iu ny chng t s t thy phn
ca tht chm, tht s lu mm. mu 2, 3, 4, lng nit formol to thnh cao
hn (cao nht mu 4), chng t enzyme thy phn nhanh cc protein tht v
vy tht mau mm. Thi gian v nng enzyme cng tng th tc thy
phn nhanh v lng nit formol to thnh cng cao.

Bang so 4.Hm lng amoniac to thnh trong qu trnh lm mm tht

Mu
Khi lng tht (g)
Khi lng enzyme (mg)
Th tch NaOH 0,1N (ml)
Lng NH3/mu tht (g)
Lung NH3/kg tht (g)

1
14,009
0
9,9
132,6.10-4
0,948

2
14,686
139,53
9,55
128.10-4
0,873

3
14,690
140,06
9,4
126.10-4
0,859

4
14,836
140,09
9,25
124,3.10-4
0,84

Ghi ch:
Mu 1: mu tht c r ng
Mu 2: mu tht c p enzyme trong thi gian 3 pht
Mu 3: mu tht c p enzyme trong thi gian 5 pht
Mu 4: mu tht c p enzyme trong thi gian 7 pht
Nu khng b sung enzyme th lng amoniac cao nht v lng
amoniac gim dn khi tng nng enzyme ln. Nh vy, dng enzyme lm
mm tht, rt ngn thi gian ch bin, s lm tng cht lng ca thc phm
ln.
2. Kho st kh nng lm mm tht ca Bro-C
T kt qu nghin cu II.1, c th ghi nhn iu kin cn thit khi s
dng Bro-M lm mm tht l:
Lng Bro-M chim 0,95% so vi khi lng tht (tng hot tnh a
vo khong 3.000 MCU/g tht);
Thi gian p tt nht l: 5 pht;

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Chng ti tin hnh so snh kh nng lm mm tht ca Bro-C (33,6

MCU/mg) vi Bro-M (320 MCU/mg), c cht s dng l tht b dai c cng khi
lng, kch thc (dy 1 cm, rng 3 cm, di 3,3 cm), lng enzyme s dng c
tng hot tnh ng t sa bng nhau, cng thi gian p, sau th cm
quan. Kt qu c trnh by Bng s 5.

Bang so 5.So snh kh nng lm mm tht ca Bro-C vi Bro-M

Enzym

Bro-M

Khi lng tht

(g)
Din tch mt
ngoi (cm2)
Khi
lng
enzyme (mg)
Thi gian p

Bro-C

16,98

16,4

16,68

16,6

17,08

16,49

16,43

15,4

32,40

31,50

32,40

31,50

32,40

31,50

31,50

30,00

160

155

155

155

1.524

1.471

1.466

1.000

5
5
5
5
5
5
5
enzyme (pht)
Tng hot tnh 51.200 49.600 49.600 49.600 51.206 49.426 49.258
Hot
tnh/g
3.015 3.024 2.974 2.988 2.998 2.997 2.998
tht
Hot
tnh/ml/cm2

79,01

78,73

76,54

78,73

79,02

78,45

78,19

5
33.600
2.049

56,00

tht
Mm
Cm quan

Mm va, n hi, khng c v

ng

Qu mm, c v ng

nhiu,
khng
ng

Kt qu trn cho thy, khc vi Bro-M, lng Bro-C s dng thch hp

nht l 56 MCU/ml/cm2 tht. Tuy nhin, do tht mm nhiu v lng enzyme
khng ha tan ht trong dung dch nn chng ti tin hnh kho st kh nng
lm mm tht ca Bro-C nhng nng thp hn. Mi mu tht th nghim c
khi lng l 20 gram (kch thc trung bnh ca ming tht 13-3,54, din
tch mt ngoi t 38-43 cm2), kt qu c minh ha Bng s 6.

-7-

Bang so 6.Kho st mm ca tht bng phng php cm quan

1

Mu

Khi lng enzyme (gram) 1,00 0,75 0,50 0,30 0,20 0,10
im t c (*) 3,4 3,5 4,2 3,3 1,4 0,6
(*) Ch tiu nh mm ca tht b sau khi p enzym bromelin bng
phng php cm quan c quy nh nh sau:
im
mm

5
Tht mm,

4
Tht mm,

3
Tht b

2
Tht hi

1
Tht

0
Tht qu

khng r

hi b r

dai

dai

dai

Kt qu cho thy mu 3 c mm tt nht, k l mu 4. Nh vy,

lng enzyme cn thit lm mm tht tnh theo din tch mt ngoi ca
ming tht trong khong 19,5-33,2 MCU/ml dung dch/cm2.
Nh vy, enzyme bromelin thu nhn t phn ph liu ca chi ngn tuy
hot tnh thp hn Bro-M nhng hon ton c th s dng trong cng ngh lm
mm tht. B sung enzyme s rt ngn thi gian lm tht mm (c bit c
ngha i vi loi tht b dai), lng m formol sinh ra cao hn, lng m
amoniac to thnh trong qu trnh thy phn protein gy thi gim xung, ng
thi cc thnh phn dng cht trong tht khng mt i so vi cch lm mm
tht thng thng l bo qun lnh ng.
Quy trnh lm mm tht b bng ch phm enzyme bromelin thu t phn
ph liu chn ngn c m t S s 2.

-8-

Tht bo
(loai dai)
Cat lat
(1cm3,5cm4cm)

Enzyme ngoai bao: 19,5-33,2

MCU/ml dung dch E.B/cm2
dien tch mat ngoai cua
mieng tht

p 5 phut
to

Tht bo mem
S s 2. Quy trnh lm mm tht b

Tht b (loi dai) c ct lt vi kch thc c tnh nh sau: dy 11,5cm, di 4,0 cm, rng 3,5 cm. Pha dung dch lm mm tht theo cch tnh nh
sau 19,5-33,2 MCU/ml dung dch p/cm2 din tch mt ngoi ca ming tht.
Sau tht trong dung dch p trong 5 pht, ri nhng nhanh vo du
nng (nh cch ch bin b beefsteak).
ti ch dng li xc nh t l enzyme bromelin cn thit c trong
dung dch p tht. to ra gi bt lm mm tht, cn phi c nhng nghin
cu tip theo v cc gia v c phi trn vo v a ra cch ch bin ngi
tiu dng d s dng hn.

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CH THCH
(1)

Enzyme Development Corporation

(2)

Nguyn Ln Dng v tp th. Mt s phng php nghin cu vi sinh

hc. Tp 3. NXB KHKT, 1978

TI LIU THAM KHO

1. Nguyn c Lng, L Th Thanh Mai v tp th (2004): Nghin cu x l

ph liu nh my hp da sn xut enzyme bromelin, phn sinh hc
v thc n gia sc. ti nghim thu cp Thnh ph. Nghim thu nm
2004.
2. B Nng nghip v Pht trin Nng thn (2002): Bo co tnh hnh sn
xut v tiu th da. Phng php pht trin v gii php thc hin. H
Ni.
3. L Th Thoa v tp th (1979) : Bo co tng hp nghin cu v bromelin.
4. Nguyn c Lng, Cao Cng (2003): Th nghim Ha Sinh hc. NXB
HQG TPHCM.
5. Phm Th Trn Chu, Phm Tin Ho, Nguyn Th Bo (1987) : Thnh
phn v mt s tnh cht ca ch phm bromelain chi ngn da ty
(Ananas Comosus L. Group Queen)
6. Nguyn nh Huyn v tp th (1994) : Nghin cu v sn xut enzyme
bromelin. ti nghim thu cp B. M s B91-07-03. Nghim thu ngy
09/9/1994.
7. Thnh, Nguyn Xun Thng (1985) : Phng php tch nhanh
bromelain t b da
8. L Th Thanh Mai (1997) : Lun n PTS khoa hc Sinh hc Nghin cu v
enzyme bromelin v con ng ng dng ca chng. Trng i hc Khoa
hc T nhin TPHCM
9. Phm Th Trn Chu, Phan Tin Ha, Nguyn Th Bo (1987): Thnh phn
v mt s tnh cht ca chi ngn da Ty (Ananas Comosus L.-group
Queen). Tp ch sinh hc, 9 (4): 3-9.

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10. Food Engineering Intl., June 1977 : Taiwan pineapples create

food

enzyme.
11. Helmut Uhlig (2000): Industrial enzymes and their applications. John Wiley
and Sons, Inc, Newyork.
12. Anthony J. Cichoke, D.C (1998): Bromelin: The active enzyme that helps us
make the most of what we eat. Keats Publishing, Inc. New Cacaac,
Connecticut.
13. Anthony J. Cichoke (1998): The complete book of enzyme therapy. Avery,
a member of Penguin Putnam Inc..

14. http://www.dole-platation.com: Dole Plantation Hawaii

TM TT
Phn trng ca chi ngn da - ph liu ca qu trnh sn xut da ng hp c th c dng nh l ngun cung cp enzyme bromelin (Bro-C). Chng c
th c dng lm mm tht. Kt qu nghin cu xc nh iu kin ti
u khi dng enzyme bromelin chi lm mm tht b: Kch thc ming tht b
(cao x di x rng) : 1-1,5 cm x 4,0 cm x 3,5 cm; Dung dch enzyme: 19,5-33,2
MCU/ml/cm2; Gi ming tht b trong dung dch enzyme trong 5 pht.
SUMMARY

The white upper parts of pineapples used to be a scrap in the process of

pineapple canning - can now be a source for bromelian (Bro-C) to tenderize
meat. The research results shows that the optimal conditions for ensuring beef
tenderness are:
- Beef size (H LW): 1-1,5 cm x 4,0 cm x 3,5 cm.
- Enzyme solution: 19,5-33,2 MCU/ml/cm2
- Meat is marinated in the enzyme solution for 5 minutes.

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- 12 -