Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27

TRCH LY ENZYME BROMELAIN T PH PHM KHM CU C-HU GIANG

Nguyn Vn Thnh1, Nguyn Minh Thy2, L H Ny3 v L Trung Hiu4

1
Vin Nghin cu v Pht trin Cng ngh Sinh hc, Trng i hc Cn Th
2
Khoa Nng nghip v Sinh hc ng dng, Trng i hc Cn Th
3
Sinh vin Sinh hc K35, Trng i hc Cn Th
4
Sinh vin Cng ngh Sinh hc K35, Trng i hc Cn Th

Thng tin chung:
Ngy nhn: 20/03/2013
Ngy chp nhn: 30/10/2013
Title:
Extraction of bromelain from
Cau Duc - Hau Giang
pineapple wastes
T kha:
Bromelain thn, kt ta
sulfate ammonium, ph phm
khm Cu c- Hu Giang,
sy chn khng
Keywords:
Ammonium sulfate
precipitation, Cau Duc - Hau
Giang pineapple wastes, stem
bromelain, vaccuum drying
ABSTRACT
Extraction of enzyme bromelain from non-used part of pineapple from
agriculture and food processing industries can give a profit for adding
value to these by products, on the other hand it can also minimize the
environmental pollution. In this study, the factors affecting on bromelain
extraction and preservation were explored. The results showed that among
different part of pineapple wastes (stem, leave, fruit) stem was the most
appropriate subject for enzyme production. The enzyme was precipitated
by 70% ammonium sulfate saturation at 28C with the protein yield
69.52%. Bromelain powder was obtained by vacuum drying for 24 hours
with the moisture content of 1.87% and specific activity of 12.29 TU/mg.
The enzymatic activity of the product was almost stabilised for twelve
weeks when it was stored in glass bottles at 4oC.
TM TT
Tch chit enzyme bromelain t ph phm khm trong cng nghip ch
bin thc phm chuyn chng t ph phm thnh sn phm c gi tr,
mt khc cng nhm lm gim thiu nhim mi trng. Vi mc tiu
trn, trong nghin cu ny cc tc nhn nh hng n trch ly v bo
qun enzyme bromelain c kho st. Kt qu nghin cu cho thy
trong s nhng phn ph phm (thn, l, tri) thn khm l ngun c cht
thch hp sn xut enzyme bromelain. Ch phm bromelain c kt
ta vi ammonium sulfate 70%, nhit 28oC, cho sn lng 69,52%
protein. Bt bromelain thu c bi sy chn khng trong 24 gi, m
t 1,87% v hot lc l 12,29 (TU/mg). Bt enzyme thu c nn bo
qun nhit lnh (4oC) trong chai thy tinh, c hot tnh n nh trong
12 tun.

1 GII THIU khm nh: li, v, thn, chi v l. Phn ph ph

Bromelain l tn gi chung cho nhm enzyme
thc vt cha nhm sulfuhydryl, c kh nng phn
gii protein c thu nhn t h Bromeliaceae, c
bit l cy khm (thn v tri). Bromelain c
nhiu tc dng trong y hc v trong ch bin thc
phm. Bromelain c mt trong ph ph phm ca
phm ny chim mt t l ln ca lng khm
nguyn liu a vo ch bin khong 70% (Li Th
Ngc H, 2009).
Theo thng k nm 2009, Hu Giang c 1.552
ha t chuyn canh khm, tp trung nhiu thnh
ph V Thanh v huyn Long M. Trong nhng

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27

nm gn y, khm Cu c c mt trong h chit v tinh sch enzyme bromelain t ph phm

thng siu th ng bng sng Cu Long v ton
quc. Khm Cu c thuc ging Queen, ngun
gc Thi Lan, thuc tiu nhm Queen c in.
Cy khm Cu c cao trn 1 mt, trng lng tri
khong 1,5-2 kg, nng sut trung bnh 20 tn/ha.
Cng vi s pht trin ca ngnh sn xut v ch
bin khm th lng ph ph phm cng tng theo.
cc nng trng trng khm, cy v l khm b
b kh trn t hoc c vi lm phn bn. cc
nh my ch bin rau qu, phn ln ph phm
khm c a ra bi rc gy nhim mi trng.
T y cho thy rng vic sn xut khm to ra
mt ngun ph ph phm rt ln c cha enzyme
bromelain. Do vy, mc tiu nghin cu nhm tch
ca khm nhm tn dng ph ph phm ca khm,
nng cao gi tr kinh t ca cy khm, trch enzyme
bromelain l cn thit nhm phc v hiu qu cho
qu trnh ch bin thc phm v dc phm v
ng thi lm gim c nhim mi trng.
2 PHNG PHP NGHIN CU
2.1 Phng tin nghin cu
Nghin cu c thc hin ti phng th
nghim Sinh Ha Thc phm, Vin Nghin cu v
Pht trin Cng ngh Sinh hc, Trng i hc
Cn Th. Ph ph phm thn, l v tri khm ph
phm (Hnh 1) c thu nhn t x Ha Tin,
thnh ph V Thanh, tnh Hu Giang.

A B C
Hnh 1: Nguyn liu th nghim: thn khm (A), l khm (B) v tri khm ph phm (C)

Ha cht th nghim Xc nh hot tnh v hm lng protein t

Aceton, (NH4)2SO4, Coomassie brilliant blue G
(n ), Tris buffer, Hydroxide natri (NaOH)
(Trung Quc), Acid chlohydric (HCl) (Trung
Quc), Casein (Trung Quc), L- Tyrosine
(C9H11O7.H2O) (n )
2.2 Ni dung v phng php nghin cu
2.2.1 Th nghim 1: Th nghim s b trch ly
enzyme bromelain t cc ngun ph ph phm
(thn, l v tri)
Th nghim c b tr hon ton ngu nhin
vi 1 nhn t gm ba ngun nguyn liu, c lp
li 3 ln v tin hnh nh sau:
Thu nhn 100 g thn khm, 100 g l khm
v 100 g tri khm ph phm ti cc vn khm
x Ha Tin V Thanh.
X l v thu dch: thn khm, l khm v
tri khm ph phm ra sch bi bn, ct thnh
ming v xay nhuyn, thu c dch. Dch thu
c tip tc ly tm lnh 8C, 7000 vng/pht
trong 15 pht). Loi b cc tp cht b ta bm
di y bnh ly tm v ch thu dch trong.
dch trong thu c bng phng php Kunitz
(1974) ci tin v phng php Bradford (1976).
Th nghim c tin hnh ring bit i
vi thn, l v tri khm ph phm v lp li 3 ln.
T chn ra ngun nguyn liu cho hiu sut
v hot tnh ti u s dng cho th nghim
tip theo.
Ch tiu theo di
Hot tnh tng ca enzyme bromelain (TU)
bng phng php Kunitz ci tin.
Hm lng protein tng s (mg) bng
phng php Bradford.
Hot tnh ring (TU/mg protein).
2.2.2 Th nghim 2: Kho st nh hng ca
nng ammonium sulfate (SA) v nhit kt ta
Th nghim c b tr hon ton ngu nhin
vi 2 nhn t v 3 ln lp li.
Nng ammonium sulfate (SA): 60-80%, cch
nhau 10%.
Nhit kt ta: nhit thng (28oC) v
nhit lnh (4oC).

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27

Thc hin th nghim: nguyn liu c nghin Nhit bo qun: nhit phng (25oC), 4oC
p, lc b b thu dch lc, ly tm lnh 15000 v -20oC.
vng/pht trong 15 pht nhit 8oC v thu dch Dng c bo qun: chai thy tinh, bao PE ghp
ly tm. Sau , cho tc nhn kt ta vo vi cc m thng.
nng nh b tr th nghim. Sau khi kt ta,
tin hnh ly tm lnh hn hp thu ta. Sy kh Cn mi mu khong 2 g bt cho vo ti PE v
ta v thu hi ch phm enzyme bromelain th. 2 g bt cho vo chai thy tinh xc nh hot
tnh trong 3 thng (12 tun). Bo qun trong iu
2.2.3 Th nghim 3: Kho st nh hng ca
kin trnh sng cc nhit v xc nh hot tnh
cc phng thc sy
enzyme sau hai tun bo qun.
Sau khi chn c nng tc nhn kt ta v
Cc ch tiu theo di cc th nghim:
nhit kt ta t th nghim 1, thc hin kho st
nh hng ca phng thc sy n hot tnh ring Lng enzyme bromelain th thu
v hiu sut thu hi hot tnh bromelain th. Th c (mg/ml) (s dng cn in t chnh xc
nghim c b tr theo th thc hon ton ngu 0,0001 g).
nhin vi 1 nhn t v 3 ln lp li. Hot tnh ring ca enzyme bng phng
Phng php sy: sy thng hoa (-20 C), sy o php Kunitz (1974) ci tin.
phun (100oC), sy chn khng (30oC) v sy bng Hiu sut thu hi enzyme bromelain (%).
kh lnh kh (4oC). Hiu sut thu hi hot tnh tng enzyme
T 2 kg thn khm ban u, ly tm ly dch v bromelain (%).
kt ta bng amonnium sulfate 70%. Sau khi kt X l kt qu
ta, ly tm lnh hn hp loi b dch v thu ta.
S dng chng trnh thng k Statgraphics.
Ta thu c a vo qu trnh sy theo cc
phng php c cp trn v thu hi 3 KT QU THO LUN
enzyme bromelain th. 3.1 Cht lng v khi lng enzyme
2.2.4 Th nghim 4: Kho st nh hng ca bromelain trch ly t cc ngun ph phm khm
iu kin bo qun ch phm enzyme bromelain th
S khc bit hot tnh v hm lng protein ca
Th nghim c b tr theo th thc hon ton enzyme bromelain t cc ngun ph phm khm
ngu nhin vi 2 nhn t v 3 ln lp li. c th hin Bng 1.
Bng 1: Kt qu phn tch khi lng v cht lng bromelain t cc ngun ph phm khm
Th tch dch Hm lng
Ngun ph phm Khi Hot tnh Hot tnh
chit enzyme th protein tng
khm lng (g) tng (TU) ring (TU/mg)
(ml) (mg)
Thn khm 100 55 4787,1c 360,43c 13,38c
Tri khm ph phm 100 65 1191,4b 119,80b 9,95b
L khm 100 40 444,5a 65,20a 6,82a
S liu trong bng l trung bnh ba ln lp li. Trong cng mt ct cc s liu i km ch ci ging nhau th khc bit
khng c ngha thng k tin cy 95%
Kt qu cho thy thn khm c hot tnh tng cc nghin cu trc, bromelain c nhiu trong
(4787,1 TU) v hot tnh ring (13,378 TU/mg) thn v tri hn so vi l. V th thn v tri c
ca enzyme bromelain l cao nht, tip l tri th s dng trch ly enzyme bromelain. Enzyme
(1191,4 TU; 9,948 TU/mg) v thp nht th hin bromelain thn v tri c im khc nhau, c
l (444,5 TU; 6,82 TU/mg). Kt qu nghin cu l do c cu thnh t hai nhm protein
ph hp vi Larrauri et al. (2004) v Nguyn c khc nhau (Ota et al., 1985). Theo Tran (2006),
Lng (2004) v hot tnh c hiu ca bromelain bromelain thn gm hn hp bn loi cystein-
tri v thn trn c cht casein, cho thy bromelain protease nn khc vi bromelain tri ch c mt
thn cao hn bromelain tri. Bn cnh , l khm loi duy nht. Nh vy thn khm l ngun thch
rt cng v c gai, thch hp cho qu trnh chua hp cho trch ly enzyme bromelain thng mi c
lm thc n cho gia sc hoc lm phn bn. Theo gi tr kinh t cao.

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27

3.2 nh hng ca nng tc nhn kt tng nhng cng ng thi tng s hin din mui
ta ammonium sulfate v nhit kt ta n trong ch phm. nghim thc 70% ammonium
cht lng v khi lng bt enzyme bromelain sulfate (SA) cho kt qu hot tnh ring v hiu
3.2.1 Nng tc nhn kt ta sut thu hi hot tnh tng l tt nht (12,36 TU/mg
v 72,59%, tng ng) v c s khc bit ngha
S kt ta bromelain di tc dng mui kim
( tin cy 95%) so vi nghim thc 60% SA (hot
cho thy nh hng ca cc nng ammonium
tnh ring v hiu sut thu hi hot tnh tng l
sulfate (t 6080%) n s kt ta enzyme
11,83 TU/mg v 72,14%, tng ng) v nghim
bromelain (Bng 2 v 3).
thc 80% SA (11,35 TU/mg v 78,01%) nhit
Khi nng mui s dng cng cao, lng kt 28oC.
ta cng tng v lng protein trong kt ta cng
Bng 2: nh hng ca nng mui ammonium sulfate nhit 28oC n s kt ta enzyme
bromelain
Nng HSTH hot tnh HSTH protein Hot tnh ring Khi lng bt enzyme
(NH4)2SO4 (%) tng (%) (%) (TU/mg) (g/0,5kg nguyn liu)
60 68,33a 54,89a 11,83ab 7,76a
70 72,59b 56,16b 12,36b 9,96b
a
80 66,96 56,02b 11,35a 11,74c
S liu trong bng l trung bnh ba ln lp li. Trong cng mt ct cc s liu c ch ci ging nhau th khc bit khng
c ngha thng k tin cy 95%. HSTH: Hiu sut thu hi
Bng 3: nh hng ca nng mui ammonium sulfate nhit 4oC n s kt ta enzyme
bromelain
Nng HSTH hot tnh HSTH protein Hot tnh ring Khi lng bt enzyme
(NH4)2SO4 (%) tng (%) (%) (TU/mg) (g/0,5 kg nguyn liu)
60 72,14a 45,37a 15,00b 8,29a
70 82,59b 59,53b 13,09a 10,07b
80 78,01ab 60,75b 12,40a 12,14c
S liu trong bng l trung bnh ba ln lp li. Trong cng mt ct cc s liu c ch ci ging nhau th khc bit khng
c ngha thng k tin cy 95% HSTH: Hiu sut thu hi
Khi thc hin kt ta 4oC th hot tnh ring nh Huyn et al. (1994), khi kt ta bromelain tri
ca nghim thc 70% ammonium sulfate (13,09 bng ammonium sulfate cc nng t 30 n
TU/mg) thp hn c ngha so vi nghim thc 100% cng kt lun nng 70% ammonium
60% SA (15,00 TU/mg) nhng khng khc bit c sulfate cho hiu sut thu hi hot tnh cao nht.
ngha so vi nghim thc 80% SA (12,40 Dng Th Hng Giang et al. (2002) nghin
TU/mg). Tuy nhin, hiu sut thu hi hot tnh cu nh gi cc phng php trch ly bromelin t
tng nghim thc 70% SA (82,59%) cao hn nc khm th bng aceton v ammonium sulfate
nghim thc 60% SA (68,33%) v nghim thc kt lun rng phng php trch ly bromelin
80% SA (78,01%). Kt qu t c nh vy c bng ammonium sulfate theo t l 70% bo ha thu
th do nng mui thp, cha kh nng kt nhn bromelain th cho hiu sut thu hi kh cao
ta nhiu protein. Khi tng lng mui ti nng 92% so vi hot tnh ban u. T kt qu th
thch hp lm kt ta hu ht cc protein v trong nghim v cng b ca cc tc gi trn cho thy kt
c s hin din ca bromelain. nng mui ta bromelin vi 70% SA l thch hp v c th
cao hn 70% bo ha, lc in tch tng ln, s c p dng cho hot ng kho st nghim
lng ion kt hp vi protein nhiu, lc tnh in thc tip theo.
c to thnh s lm cho phn t protein b xoy, 3.2.2 Nhit kt ta
vn v dn n s bin tnh (Nguyn nh Huyn
et al.,1994). Nh vy, nng mui ammonium Thc nghim cho thy nhit nh hng quan
sulfate 70% bo ha l kt ta ht bromelain trng n qu trnh trch ly bromelain (Bng 4).
c trong dch thn khm. Theo kt qu ca Nguyn

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27

Bng 4: nh hng ca nhit n s kt ta enzyme bromelain

Nhit HSTH hot tnh HSTH protein Hot tnh ring Hot tnh TU/g
(oC) tng (%) (%) (TU/mg) enzyme
4 82,59b 59,53b 13,09b 2751,65b
28 72,59a 56,16a 12,36a 2422,30a
S liu trong bng l trung bnh ba ln lp li. Trong cng mt ct cc s liu c ch ci ging nhau th khc bit khng
c ngha thng k tin cy 95%. HSTH: Hiu sut thu hi
Kt qu cho thy khi kt ta nhit 28oC th v 28oC thc hin kho st cho th nghim
hot tnh ring v hiu sut thu hi hot tnh tip theo.
bromelain l 12,36 TU/mg v 72,59%, tng ng, 3.3 nh hng ca phng thc sy n
thp hn c ngha ( tin cy 95%) so vi hot tnh ring, hiu sut thu hi hot tnh v
nghim thc kt ta nhit 4oC (hot tnh ring khi lng bt enzyme bromelain
v hiu sut thu hi hot tnh bromelain tng ng
l 13,09 TU/mg v 82,59%). Nh vy khi thc Nhit sy v thi gian sy l hai yu t quan
hin kt ta 4oC vi tc nhn ta ammonium trng nh hng n hot tnh ca ch phm
sulfate th hot tnh ca enzyme lun lun tt hn enzyme th. Sy nhit cao c th lm mt hot
nhit 28oC bt k nng mui no. nhit tnh ca enzyme do protein b bin tnh. Tuy nhin,
lnh (4oC), qu trnh kt ta protein v kh nng khi sy nhit thp th thi gian sy ko di v
hn ch s bin tnh din ra tt hn. Kt qu thu trong thi gian ny vi s hin din ca m cao
c cng tng t kt qu nghin cu c th s t phn ca cc protease s din ra lm
cng b ca Nguyn nh Huyn et al. (1994). enzyme b mt hot tnh. Kt qu cho thy khi sy
Nhm tc gi cng cho thy rng mui ammonium ng kh ch phm enzyme bromelain th trong 12
sulfate s dng cho kt ta bromelain cng cho gi cho hiu sut thu hi hot tnh v hot tnh
hot tnh ca ch phm trch ly nhit 4oC ring ca bt enzyme th cao nht (87,55% v
thng cao hn nhit 27oC. Tuy nhin, nhn 12,96 TU/mg, tng ng) v th hin s khc bit
chung khi ly trch nhit phng (28oC), hot ngha so vi 3 nghim thc sy cn li (Bng 5).
tnh tng ca enzyme gim 10% so ly trch nhit Tuy nhin, khng c s khc bit ngha v hiu
o
4 C v vn mc chp nhn c. Mt khc, t sut thu hi hot tnh v hot tnh ring ca bt
thc nghim cho thy qu trnh kt ta nhit enzyme th gia cc phng thc: sy chn khng
o
4 C din ra kh khn, i hi phi c thit b trong 24 gi (72,02% v 12,23 TU/mg), sy kh
lm lnh nn rt tn km v thao tc phc tp nn lnh kh trong 48 gi (69,52% v 12,29 TU/mg) v
chi ph sn xut cao. Nh vy, nhm hon thin phng thc sy phun vi cht bo v l mannitol
phng php s dng mui ammonium sulfate lm 5% (Nguyn Ph Th v Dng Th Hng Giang,
tc nhn kt ta bromelain t thn khm, c th 2011) trong 1 gi (40,82% v 11,51 TU/mg).
chn nng v nhit kt ta tng ng l 70%
Bng 5: nh hng phng thc sy n hot tnh enzyme bromelain
HSTH hot HSTH Hot tnh Khi lng bt
m
Phng thc sy tnh tng protein ring enzyme (g/0,5 kg
(%)
(%) (%) TU/mg nguyn liu)
Sy phun* (100oC) 40,82a 34,65a 11,51a 2,52a 4,96a
Sy chn khng (30oC ) 72,02b 57,50b 12,23b 1,87a 6,03b
Sy kh lnh kh (4oC) 69,52b 55,08b 12,29b 3,57b 6,28c
Sy ng kh (-20oC) 87,55c 66,67c 12,96c 1,82a 5,85b
S liu trong bng l trung bnh ba ln lp li. Trong cng mt ct cc s liu c ch ci ging nhau th khc bit khng
c ngha thng k tin cy 95%. HSTH: Hiu sut thu hi. * Mu enzyme kt ta thu c t 500 g mu thn khm
sau trch ly c x l bi ha tan vo trong 210 ml nc ct v thm vo mannitol 5%, tin hnh sy phun
Nh vy, hot tnh enzyme n nh nht khi khng lm hng enzyme. Nu nhit thng d
thc hin sy bng phng php ng kh. Vi lm enzyme bin tnh th phng php ng kh
phng php ny, nc tch ra khi nguyn liu gip enzyme gi c tnh cht ha l v c hot
bng cch chuyn t trng thi rn () sang trng tnh. Li Th Ngc H (2009) so snh hiu qu ca
thi kh m khng qua giai on lng (nc) phng php sy phun v sy ng kh bromelain
-40oC. Nh vy nc c a ra khi enzyme m

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27
cho kt qu hot tnh c gi li sau hai qu trnh
ny l 96% v 73%, tng ng.
Sy chn khng cng l mt bin php lm
kh ta bromelain n m an ton v c th
bo qun trong thi gian di. Tuy nhin so vi
sy ng kh, sy chn khng c tin hnh
nhit cao hn nhiu (30oC) nn hot tnh ca
bromelain sau khi sy thp hn. Tuy nhin khi so
snh gia sy chn khng vi phng php sy
bng kh lnh kh nhit 4oC th cc gi tr thu
nhn c khng th hin s khc bit ngha (
tin cy 95%). iu ny c th l do bromelain l
mt enzyme bn nhit trong khong nhit 50
60oC, nn nhit khong 30oC vn gi c
hot tnh tng i tt nh nhit lnh (4oC).
Khi s dng phng php sy phun, hot tnh
enzyme bin tnh kh nhiu do bn nhit ca
bromelain km hn papain. 70oC, bromelain b
bin tnh nhanh gp 20 ln papain 75oC, tu
thuc c cht m nhit ti u khc nhau
(Nguyn Trng Cn et al., 1998).
Kt qu th hin ng thi Bng 5 cho thy
khi lng bt enzyme thu c t phng php
sy bng khng kh lnh kh l cao nht (6,28g) v
th hin s khc bit ngha so vi ba nghim thc
sy cn li. Tuy nhin, khng c s khc bit
ngha gia nghim thc sy chn khng (6,03g) v
nghim thc sy ng kh (5,85g). Khi lng bt
enzyme thu c t phng php sy phun c khi
lng thp nht (4,96g). iu ny c lin quan n
m ca sn phm, sy bng khng kh lnh ch
phm t m 3,57%, cao hn m ch phm
thu c t phng php sy chn khng (1,78%)
v sn phm sy ng kh (1,82%). Ch phm
bromelain thu c t phng php sy phun c
khi lng thp nht, c l do trong qu trnh sy,
enzyme b tht thot theo khng kh nng.
Ngoi ra, thi gian sy t cc phng php sy
cng khc nhau. Vi cng lng mu nh nhau,
sy n m bo qun an ton ca ch phm
enzyme (<8%) th phng php sy chn khng
mt khong 24 gi, trong khi sy bng kh lnh kh
phi mt 48 gi (vi yu cu thit b gi lnh 4oC),
khng kh phi c m thp v vic lun chuyn
kh thc hin. Vi phng php sy ng kh th
ngoi thi gian sy tng i ngn (10 gi),
phng php ny cn i hi mu trc khi a
vo sy phi c lm lnh ng n -20oC, vn
hnh thit b sy ng kh cng kh phc tp v
t tin nn trong thc t cha c s dng
rng ri. Vi phng php sy phun th thi gian
sy ngn nht (1 gi), d sn xut quy m cng
nghip tuy nhin lng enzyme tht thot ln v d
mt hot tnh do sy nhit cao. Nh vy, c
th chn phng php chn khng cho qu trnh
sy bt enzyme bromelain th cho th nghim
tip theo.
3.4 nh hng ca iu kin bo qun n
s bin i cht lng enzyme bromelain th
Enzyme bromelain th c bo qun cc
nhit 25, 4, -20oC. Kt qu xc nh hot tnh
ring ca enzyme cho thy nhit 25oC, hot
tnh enzyme gim nhanh nht (19,9%) v gim t
hn i vi enzyme c bo qun 4oC (8,25%)
v gim t nht khi c bo qun -20oC (4,96%)
trong bn thng bo qun (Hnh 2) c s khc bit
ngha vi tin cy 95% gia cc nhit ny.
Enzyme bromelain c bo qun trong chai thy
tinh v bao PE. Kt qu xc nh hot tnh ring
cho thy enzyme bo qun trong chai thy tinh tt
hn (hot tnh 11,59 TU/mg) cao hn (c ngha)
so vi hot tnh enzyme cha ng trong bao PE
(11,34 TU/mg) (Bng 6).
Kho st hot tnh enzyme bromelain thnh
phm cho thy hot tnh tng v hot tnh ring
ca lai ny cao hn (2165,12 TU v 12,51
TU/mg) v th hin s khc bit ngha so vi ch
phm enzyme bromelain thng mi (1929,1 TU
v 7,01 TU/mg) (Bng 7).
14
Hot tnh ring (TU/mg)
12
10
8
6
4
2
0
0 15 30 45 60 75 90 105
Thi gian bo qun (ngy)
25oC 4oC -20oC
Hnh 2: Bin i tnh c hiu ca bromelain
theo nhit v thi gian bo qun
Bng 6: nh hng ca dng c cha n hot
tnh enzyme trong qu trnh bo qun
Dng c cha Hot tnh (TU/mg)
Bao PE 11,34a
Chai thy tinh 11,59b
S liu trong bng l trung bnh ba ln lp li. Trong
cng mt ct cc s liu c ch ci ging nhau th khc
bit khng c ngha thng k tin cy 95%
26
Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 28 (2013): 21-27
Bng 7: So snh hot tnh ca bt enzyme thnh
phm v bt enzyme thng mi
Hot Hot tnh
Loi bt
tnh tng
bromelain
(TU)
Bt enzyme thnh
2165,12b
phm
Bt enzyme thng
1929,10a
mi (Merck)
S liu trong bng l trung bnh ba ln lp li. Trong
cng mt ct cc s liu c ch ci ging nhau th khc
bit khng c ngha thng k tin cy 95%
4 KT LUN
Kt qu thu nhn c t nghin cu cho thy
thn khm l ngun nguyn liu thch hp trch
ly bt enzyme bromelain. C th thc hin tt qu
trnh kt ta enzyme bromelain bng ammonium
sulfate 70% 28oC. Hiu sut thu hi enzyme tt
nht vi phng thc sy chn khng trong 24 gi,
ch phm c mu trng ng, hiu sut thu hi hot
tnh tng l 69,52%, hot lc 12,29 (TU/mg) v
m 1,87%. Bt enzyme thnh phm cn bo qun
trong chai thy tinh nhit lnh (khong 4oC).
Quy trnh trch ly ch phm enzyme bromelain
theo phng php kt ta bng ammonium sulfate
c ngh sau:
Thn khm
Nghin, p
Lc B
Ly tm 1 (7000 v/pht) B
Kt ta bng ammonium sulfate 70%
(nhit phng 28oC)
Ly tm 2 (7000 v/pht) Dch
Ra ta bng acetone lnh
Sy chn khng trong 24 gi
Bromelain th
Bo qun trong chai thy tinh 4oC
Hnh 3: Quy trnh trch ly enzyme bromelain t
thn khm ph phm
TI LIU THAM KHO
1. Bradford M. M, 1976, A rapid and sensitive
mocrogam quanlities of protein utilizing the
ring principle of protein dye binding, Analytical
(TU/mg) Biochemistry, 72, 248-254.
2. Dng Th Hng Giang, L Thanh Hng,
12,51b
V Vn Song Ton, Sonia Beeckmans,
Edilbert Van Driessche v Trn Phc
7,01a
ng, 2002. nh gi cc phng php trch
ly bromelin t nc khm th, Tp ch Khoa
hc nm 2002, Trng i hc Cn Th.
3. Kunitz M, 1974. Determination of proteolytic
activity by the casein digestion method.
Journal of General Physiology, 30, 291.
4. Li Th Ngc H, 2009. Nghin cu tch v
to ch phm enzyme bromelain t ph ph
phm da, Tp ch Khoa hc v Pht trin
nm 2009 - tp 7, s 2:203-211, Trng i
hc Nng nghip H Ni.
5. Larrauri J.A., Ruperez P. and Calixto F. S.,
1997. Pineapple shell as a source of dietary
fiber with associated polyphenols. Journal
of Agricultural and Food Chemistry, 45:
4028-4031.
6. Nguyn nh Huyn, H i Quc, Lm Th
Kim Chu, L Th Thanh Mai, 1994. Nghin
cu v sn xut enzyme bromelain. ti
nghim thu cp B. M s B91-07-03.
7. Nguyn c Lng, 2004. Cng ngh enzyme,
NXB HQG Thnh ph H Ch Minh.
8. Nguyn Ph Th v Dng Th Hng
Giang, 2011. Nghin cu qui trnh iu ch
bt enzyme papain th t nha u . Tp
ch Khoa hc Trng i hc Cn Th.
9. Nguyn Trng Cn, Nguyn Th Hin,
Th Giang, Trn Th Luyn, 1998. Gio
trnh Cng ngh enzyme, NXB Nng
nghip, H Ni.
10. Ota S., Muta E., Katahira Y., Okamoto Y.,
1985. Reinvestigation of fractionation and
some properties of the proteolytically active
components of stem and fruit bromelains.
Journal of Biochemistry (Tokyo) 98: 219-228.
11. Tran A.V., 2006. Chemical analysis and
pulping study of pineapple crown leaves,
Industrial crops and products, 24: 66-74.
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