- 1 -

PHN M U

H tiu khng ch l loi ht gia v quen thuc trong gia nh m cn l loi
cy cng nghip mang li gi tr kinh t cao cho ngi nng dn. Hng nm em
li ngun thu ngoi t ln cho t nc. Nhng nm gn y, cy tiu tr thnh
mt trong nhng loi cy trng ch lc ca cy cng nghip, khng ch gip ngi
dn xa i gim ngho m cn l cy mang li cuc sng giu c cho nhiu h
nng dn. V th vic trng, ch bin v xut khu h tiu c nc ta u tin
ch trng u t v pht trin.
Mt khc yu cu ca ngi tiu dng ngy cng cao v cht lng sn
phm, do hin nay vic sn xut tiu s cn mang tnh th cng, c s dng ha
cht nh H
2
O
2
, CaO, HCl

nh hng n sc khe ngi tiu dng, nc thi

sau khi sn xut tiu s c cc cht mu, mi v tnh st khun mnh nn tiu dit
h vi sinh vt gy nhim mi trng, mt cn bng sinh thi.
Lu nay, ngi trng tiu thng c thi quen, sau khi thu hoch ch phi kh,
sau em vo my ch loi cung v l ri bn cho cc thng li, hoc sn
xut bng cch ngm nc, ch ra v s dng ha cht lm tiu s. Cha ai
thc s quan tm n khu phi, s ch ng tiu chun, nn ht tiu en thng b
nhim bi, vi khun, nm mc... gy tc hi cho ngi tiu dng, gi li thp, nht
l khng c gi tr xut khu. Trong khi tiu trng (tiu s) va m bo cc tiu
chun v sinh an ton thc phm, li c gi tr xut khu kh cao (gp 2-2,5 ln tiu
en)
V hin nay do vn v sinh an ton thc phm v nhim mi trng nn
vic hn ch s dng ha cht v s dng cc sn phm c sn xut t cc enzym
vi sinh vt c khuyn khch. Rt nhiu chng vi sinh vt c s dng nh
Aspergillus niger, Botrytis cinerea, Fusarium moniliforme, Rhizopus stolonifer,
Trichoderma viride, Neurospora crassa, Altenaria tenuis, Aspergillus fumigatus,
Aspergillus flavus, Celluvibrio gilvus, Fusarium culmorum, Trichoderma koningi
- 2 -
Hin nay trn th gii c ti nghin cu ca Vaidyanatha Lyer
Thankamani, Raghavan Nair Giridhar Khoa Cng ngh sinh hc, Trng i hc
Kariavattom ca Kerala ln men sn xut tiu trng s dng cc chng vi khun
Bacillus mycoides, Bacillus licheniformis, Bacillus brevis vi kt qu nh lng
piperine 4%.
Vit Nam gp phn nghin cu s dng ch phm sinh hc ln men sn
xut tiu trng vi nhng ti khoa hc ca PGS.TS Nguyn c Lng v cng
s Trng i Hc Bch Khoa thnh ph H Ch Minh nghin cu v cung cp
cc loi enzym hu c (Aspergillus niger) cng quy trnh ch bin tiu trng.
K s Bi Thanh Ha v k s V Th Ngc Thanh Khoa Cng Ngh Sinh
Hc Trng i hc Quc T - HQG p dng thnh cng ch phm sinh hc
Biocoffee-3 (A. phoenicis) t l bc v 93% v nh lng piperine 5%. Piperine
trch ly t ht tiu ln men bng ch phm Biocoffee-3 tng 21%. Ch phm sinh hc
Biopep-6 (T. piluliferum) kt qu hm lng cc cht ha tan tng 25% v lng
piperine cao 5.7% .
gp phn nghin cu lm a dng thm nhiu ch phm sinh hc v tng
hm lng piperine chng ti nghin cu ti S DNG CH PHM SINH
HC LN MEN SN XUT TIU TRNG t enzym ca vi sinh vt Aspergillus
niger, Aspergillus oryza v Bacillus polyfermenticus, Bacillus amyloliquefaciens.
Mc ch ti: S dng ch phm sinh hc (c cha vi sinh vt) tng hp
enzym pectinase v cellulase phn gii pectin, cellulose l thnh phn chnh cu
to nn v ht tiu, sn xut tiu trng (tiu s) vi hm lng piperine tng cao
cung cp cho th trng vi quy trnh sn xut thn thin vi mi trng v sn
phm vn gi c mu sc mi v t cht lng, an ton cho ngi s dng.
t c mc ch ca ti chng ti ra cc ni dung nghin cu sau:
1) Kho st m ht tiu cho qu trnh ln men sn xut tiu trng.
2) Kho st t l ch phm tt nht cho qu trnh ln men sn xut tiu trng.
3) Kho st thi gian ln men t t l bc v ht tiu cao nht.
- 3 -
4) Kim tra nh lng piperine trong ht tiu
5) Kim tra c tnh aflatoxin, ochratoxin
6) Kim tra hm lng vi sinh trong sn phm
Nu t c cc ni dung nghin cu trn th c th ng dng ti vo
quy m sn xut cng nghip cc cng ty sn xut tiu trng. ngha khoa hc
ti l s dng ch phm t vi sinh vt c ngun gc t nhin, khng s dng cht
ha hc, thi gian sn xut ngn, chi ph thp, an ton cho ngi s dng, sn phm
t cht lng cao, quy trnh sn xut thn thin vi mi trng.




















- 4 -
CHNG 1: TNG QUAN TI LIU

1.1 Gii thiu v cy h tiu
H tiu c ngun gc t n v c s dng lm gia v trong cc mn n
t nht l t 2000 nm trc cng nguyn [26]. Cy h tiu (Piper nigrum L.) thuc
h Piperaceae, b Piperales, c s nhim sc th 2n = 52 [12]. H tiu c danh php
ha hc l Piper nigrum l mt loi cy leo c hoa thuc h h tiu trng ch yu
ly qu v ht, thng dng lm gia v di dng kh hoc ti. [16]
1.1.1 Yu cu sinh thi v phn b a l ca cy h tiu.
H tiu l mt loi cy c trng ca vng nhit i m. Cc vng trng h
tiu chnh trn th gii ch yu nm vng Chu Thi Bnh Dng nh n ,
Indonesia, Malaysia, Sri Lanca, Thi Lan v Vit Nam. H tiu cng c trng
cc nc khc nh Brazil, Madagascar. Cy h tiu i hi lng ma cao, nhit
kh cao ng u v m khng kh cao, l kiu kh hu c trng ca vng
nhit i nng v m vi s thay i khng ng k v di ngy v m khng
kh trong sut nm. [12]
Cn c vo cc c tnh thc vt ca cy h tiu nh hnh dng l, phc hoa,
chm qu v.v... m ngi ta phn bit cc ging h tiu khc nhau. Trong qu trnh
canh tc lu i ti mi khu vc hnh thnh nn nhng ging h tiu a phng
nh tiu Tin Sn vng Pleiku, tiu Vnh Linh Qung Tr, tiu Lc Ninh Bnh
Phc (gm tiu s Lc Ninh v tiu trung Lc Ninh), tiu Ph Quc v.v... [12]
1.1.2 c im hnh thi ca cy h tiu
Thn: H tiu dng dy leo bng r ph. Thn trn nhn, ty ln nhng mu,
mc un o [16]. Loi thn tho mm do c phn thnh nhiu t, ti mi t c
1 l n. [12]
L: L c cung, mc so le. Hnh bu dc hoc hnh trng, di 11-15 cm,
rng 5-9 cm, gc trn, u nhn, c nhng chm trong rt m, mt trn lc sm
bng, mt di mu xm nht, gn l li r, cung l di 2-3 cm. [16]
- 5 -
Hoa: Cm hoa mc i din vi l thnh bng ngn hn l. L bc thun
di, cng ln ln vi trc bng, hoa khc gc, i khi tp tnh c bao bc bi l
bc, nh 2, bao phn hnh thn, bu thng hnh trn [16]. Hoa h tiu ra khng tp
trung m ra thnh nhiu la. Hoa t ca h tiu hnh gi, treo lng lng, di 7 -
12cm ty ging v ty iu kin chm sc. Trn hoa t c bnh qun 20-60 hoa xp
thnh hnh xon c, hoa lng tnh hay n tnh. Cc ging h tiu cho nng sut
cao thng c t l hoa lng tnh nhiu hn. [12]
Qu: Qu h tiu thuc loi qu hch, khng c cung, qu hnh cu [12].
Khi chn mu , em phi kh thnh en, ng knh 3-4 mm, ht trn. Mt ngoi
mu nu en, c nhiu vt nhn hnh vn li ni ln. nh u qu c vt ca vi
nhy nh hi ni ln, gc qu c vt so ca cung qu. Qu cng. V qu ngoi c
th bc ra c. V qu trong mu trng tro hoc mu vng nht, mt ct ngang
mu trng vng. Qu c cht bt, trong c l hng nh. Mi thm, v cay. [16]
Ma hoa qu: Thng 5-8 [16]
Thnh phn ha hc ca ht tiu: Tinh du 1,2-2%, Piperin 5-9%,
Chanvixin 2,2-6%, Cht bo 8% ,Tinh bt 36% , Tro 4% [23], [25].
Hnh 1.1: Cy tiu ang ra hoa [48] Hnh 1.2: Gi hoa v chm qu cy tiu[48]
http://www.lrctnu.edu.vn:8080/gsdl/collect/khuyenno/index/assoc/HASH72d9.dir/do
c.pdf [49]
- 6 -
Bng 1.1 Thnh phn dinh dng ht tiu [47]
1 tbsp tiu en
Nng lng 16
Nng lng t cht bo 1.89
Tng s %DV
Tng cht bo 0.21g 0%
Cht bo bo ha 0.063g 0%
Cht bo khng bo ha n 0.065g
Cht bo khng bo ha a 0.072g
ng 0.04g
Cholestreol 0mg 0%
Natri 3mg 0%
Kali 81mg 2%
Tng Carbohydrate 4.15g 1%
Cht x 1.7g 7%
Protein 0.7g 1%
Canxi 3mg 0%
St 10mg 56%
Vitamin C 2%
http://www.foodguts.com/?ingredient=black+pepper

1.1.3 Cc loi sn phm h tiu trn th trng th gii
Sn phm h tiu thng dng trn th trng th gii l tiu en v tiu trng.
Ngy nay nhiu loi sn phm c gi tr c pht trin thm t h tiu. iu
thun li l khng cn dng nguyn liu h tiu tt ch bin ra cc loi sn phm
ny. V d nh tiu lp, l loi tiu c phm cp rt km c dng sn xut ra
du tiu. n l mt nc c v tr quan trng trong vic sn xut v xut khu
cc sn phm h tiu c gi tr tng thm. [12]

- 7 -
1.1.3.1 Tiu en
Ton tri tiu bao gm v tri v ht c phi kh n m 13%. Tiu
en thnh phm c mu en vi lp v ht nhn bc bn ngoi. [12]
1.1.3.2 Du tiu
L tinh du bay hi, c chit xut t qu tiu bng phng php chng ct
hi nc. l mt hn hp lng t nhin, trong sut, c mu xanh vng n hi
xanh l cy. [12]
1.1.3.3 Oleoresin tiu
Cn gi l du nha tiu, l mt cht chit xut m c bi s chit xut
bng cc dung mi c truyn hoc chit xut nhit cao. Oleoresine l mt hn
hp tinh du, nha v cc hp cht nh piperine c bn cht l alkaloid to v cay.
Du nha tiu c y cc c trng v hng v thm cay ca tiu. [12]
1.1.3.4 Tiu bt
Ht tiu kh c nghin cc kch c khc nhau ty theo yu cu ca
ngi tiu th. Gn y cng ngh xay tiu bt nhit thp c gii thiu
trnh s mt mt cc cht thm bay hi khi nghin ht tiu. Nghin tiu nhit
thp cng loi b c vi khun v nm mc. [12]
1.1.3.5 Tiu xanh ngm nc mui
Tiu xanh ngm nc mui c ch bin t qu tiu cha chn. Sau khi
hi, tch cn thn qu tiu khi gi, trnh lm v, dp qu. Cc qu tiu (ht tiu
xanh) ny c ngm trong dung dch gim v mui gi c mu xanh t nhin
v th cht dn, xp ca ht tiu xanh. Tiu thnh phm c hng v thm ngon
c ngi tiu dng chp nhn. [12]
1.1.3.6 Tiu xanh kh nc
y l mt loi sn phm c ch bin t ht tiu xanh, bng cch x l ht
nhit cao lm v hiu s hot ng ca cc enzym lm ht tiu ha nu en.
Tiu xanh qua x l nhit sau c sy kh hay phi kh nhit c kim
sot, nh vy gi li c mu xanh t nhin nh khi thu hi. Sau khi ngm vo
nc, ht tiu s phc hi li hnh dng v mu sc gn ging nh ht tiu xanh khi
- 8 -
thu hi. Ma thu hoch tiu xanh ch din ra trong mt thi gian ngn trong nm,
trong khi nhu cu tiu dng tiu xanh sut nm. Tiu xanh kh nc c th tn
tr c trong mt nm, p ng vi nhu cu tiu dng. [12]
1.1.3.7 Tiu xanh ci tin
khc phc nhc im v th cht v hng v ca tiu xanh kh nc,
khc phc nhc im v chi ph bao b ng gi ca tiu xanh ngm mui, ngi
ta c nhng ci tin trong ch bin tiu xanh. sn xut ra loi tiu xanh ci
tin ny, ht tiu xanh c ra sch trong nc, bc tip theo l em ngm trong
nc mui 2-3 thng, x nc ri ng gi trong cc ti PE a ra th trng.
[12]
1.1.3.8 Tiu xanh ng kh
y l mt sn phm tiu xanh ho hng c ch bin bng cch lm kh
ht tiu xanh n m khong 4% nhit m 30- 40
0
C trong iu kin chn
khng. Mu sc, hng thm v th cht ca tiu xanh ng kh tt hn nhiu so
vi tiu xanh phi kh di nh sng mt tri hay c kh nc qua sy. Sn
phm ny c th gi nhit bnh thng trong phng. Khi uc lm m tr li,
sn phm ny ging nh sn phm tiu xanh mi thu hi qu trnh ch bin i hi
my mc phc tp nn sn phm tiu xanh ng kh c gi rt t. [12]
1.1.3.9 Tiu
Khi tiu chn hon ton, mu ca qu tiu chuyn t xanh sang . Mu
rt hp dn so vi mu en hay mu trng ng ca tiu trng. ch bin tiu ,
thu hi tiu khi nhiu qu tiu trn chm qu chn . Cc qu ny c tch cn
thn ra khi chm qu. Cc qu cn li c li 2-3 ngy cho ti khi chuyn sang
mu th c tip tc ch bin thnh tiu . Cc qu tiu sau khi c tch
ra khi chm tri phi ch bin trong ngy. Mu ca qu tiu c gi li bng
cch ngm qu tiu vo dung dch nc mui hay mui v gim cng vi cht bo
qun thc phm. Sau tiu c th c kh nc nh quy trnh kh nc ca tiu
xanh. Ngoi ra cn c cc sn phm khc c ch bin t tiu nh tr tiu, ko
tiu, du thm tiu, tiu dng cho hng liu m phm. [12]
- 9 -

Hnh 1.3: Tiu kh: Tiu xanh, tiu en, tiu , tiu trng [57]
http://www.uni-graz.at/~katzer/engl/Pipe_nig.html
1.1.3.10 Tiu trng
Tiu trng hay cn gi l tiu s. Qu tiu chn gi c tch lp v bn
ngoi ri phi kh. Tiu trng thnh phm ht trn nhn c mu trng ng.
a. Cch ch bin tiu trng th cng
L tiu chn gi, chm tri c hn 50% tri chn mi hi, em 2 - 3
ngy m sau tch ht, b vo bao em ngm dng nc chy, hay trong b
ngm c thay nc hng ngy. Ngm t 7-10 ngy cho n khi v nt ri, cho vo
r hay my xt k sau i ht v v phi 1-2 nng trn nong, nia n khi ht c
m 12 - 13% l c th em bo qun v tiu th. Ch bin th cng thng vi
s lng t v phi la tng chm tri rt mt thi gian v nhn cng khi thu hoch.
[12]
b. Ch bin bn cng nghip quy m nng h
Tiu trng cn c ch bin cng nghip t tiu en loi c dung trng
570g/lt. t c loi tiu en c dung trng cao dng lm nguyn liu ch bin
tiu trng th phi thu hoch lc chm tri c 2-3 qu chn . Tiu en c cht
lng cao nh ni trn em v qut k chn ht nng, em ngm nc trong
vng 7-8 ngy. Hai ba ngy thay nc mt ln, n khi v tri tiu nt ri th em
ra xay xt loi b v ht, i sch ri phi kh trn bt, sn phm c mu vng
- 10 -
ng [21]. Theo yu cu ca th trng ngi ta c th lm trng bng cch ngm
trong H
2
O
2
2% trong vng 30 pht oxy ha cht hu c v cht mu. Sau khi
lm trng tin hnh phi trn sn c lt bt, m hoc sy ht tiu nhit 50-
60
0
C trong nhiu gi lin tc ht t m 12%.-13%
Ch bin tiu en thnh tiu trng gip cc nng h tng thu nhp, tuy vy
do cha ch n vn mi trng, ti cc vng ch bin tiu trng thng xy ra
tnh trng nhim mi trng do mi hi thi bc ln t cc b ngm tiu, nc
ra ht tiu sau khi ngm trong qu trnh ch bin c cht mu, tnh st khun
mnh tiu dit h sinh thi gy nhim mi trng [12].

Hnh 1.4:B ngm tiu en [12] Hnh 1.5:My xt v ht tiu sau khi [12]
c. Sn xut tiu trng quy m cng nghip s dng ch phm sinh hc
Ch phm Biovina 9 do H Bch Khoa TP.HCM nghin cu, sn xut c
p dng sn xut tiu s ti Cng ty Olam Vit Nam, Cng ty Long Xng (Qun
5). Ch phm Biocoffee-3 (A. phoenicis) v Bipep-6 (T. piluliferum).[29], [34]
Tiu en c la chn loi b tiu khng tiu chun v tp cht, ngm
nc t m ti u sau tin hnh phi trn ch phm, thi gian ln men
khong 4 ngy sau tin hnh ch xt v ra bng nc sch nhiu ln, sy t
m 12-13% [12], [29], [34].


- 11 -
1.1.4 chn thu hoch ca ht tiu theo sn phm. [12]
Ty theo sn phm c ch bin, tiu c thu hoch cc chn khc
nhau.
Bng 1.2: chn thu hoch ca ht tiu theo sn phm.
Sn phm chn khi thu hoch
Tiu ngm nc mui/ng hp ang xanh v cn cha cng ht (vo
khong 4-5 thng sau khi ra hoa
Tiu xanh kh nc (vn gi mu
xanh)
10-15 ngy trc khi chn hon ton
Du nha tiu v Oleoresin 15-20 ngy trc khi chn
Tiu bt Chn hon ton vi ht tiu cng chc
Tiu en Chn hon ton vi ht tiu cng chc,
trn chm qu c 1-2 qu bt u chuyn
sang vng
Tiu trng Chn hon ton, trn chm qu c t nht
2-3 qu bt u chuyn sang chn
Tiu Chn hon ton, trn chm qu c nhiu
qu chn

1.1.5 Tnh hnh sn xut h tiu trn th gii v Vit Nam
1.1.5.1 Tnh hnh sn xut h tiu trn th gii
Theo c tnh Hip Hi tiu th gii, tnh hnh sn xut ht tiu nm 2009
tng 5% so vi nm trc l 305.070 tn. Vic sn xut tiu en t 246.600 tn
trong khi sn lng ht tiu trng c tnh t 58.470 tn chim 19.17% v d kin
tng lai sn lng tiu trng s tip tc tng cao. Tuy nhin, con s tiu th dao
ng xung quanh 315.000 tn v cn li tn tr trong kho l 68.700 tn. [32].

- 12 -

Biu 1.1: Tnh hnh sn xut tiu trn th gii (n v tn) (ngun IPC)[32]
Biu 1.1 m t r rng xu hng sn xut tiu ton cu t nm 1995. Sn
xut ht tiu ton cu ch khong 220.000 tn vo nm 1995 v tri qua mt thi
gian c mt s gia tng ng k t nm 1999 vi s ng gp to ln ca cc
quc gia c sn lng tiu ln nh Vit Nam, n , Malaysia, v Brazil. C c
iu ny l nh vo s ci tin trong cch qun l, ging cy trng cng vi vic
trng li tiu trong bn nm tr li y.
Nc sn xut h tiu ln trn ton th gii l Vit Nam, n , Brazil,
Indonesia, Malaysia, Trung Quc, Srilanka, Thi Lan v Singapore. Vit Nam ng
u danh sch vi th phn 34% trong sn xut ht tiu ton cu. n v tr th
hai (19%) c theo sau bi Brazil 13%, Indonesia 9%, Malaysia 8%, Trung Quc
7%, Srilanka 6 %...[32]




- 13 -

Biu 1.2 : Cc quc gia sn xut h tiu chnh trn th gii [32]
a. Vit Nam
Vit Nam quc gia sn xut v xut khu h tiu ln nht th gii. Vit Nam
bt u sn xut h tiu vo nm 1990 v bt ln l nc ng u trong mi nm
vi ng gp 95.000 tn vo sn lng tiu ton cu hin nay. nh cao l nm
2004 v nm 2006 sn lng tiu Vit Nam t trn 100.000 tn. Din tch sn xut
ht tiu ti Vit Nam dao ng khong 50.000 ha [32].


Biu 1.3: Khuynh hng sn xut tiu Vit Nam [32]

- 14 -
b. Barzil
Brazil l nc sn xut h tiu ln th 3 trn th gii. Vic sn xut h tiu
Brazil chu tht bi trong nm 2009 do thit hi v dch bnh. Sn lng thp hn
20% so vi nm 2008 v ch t 30.000 tn 2010. S lng xut khu ca Brazil
ng gn 22.000-25.000 tn.
Theo d liu ca hng nhp khu M n thng 7/2009, xut khu ca Brazil
sang M c 750 tn trong khi n l 366 tn. Brazil nm 2009 a ra
mt s cnh tranh mnh m cho n bng cch cung cp gi thp nht trong bn
nc ln xut khu Vit Nam, n , Indonesia v bao gm Brazil [32].

Biu 1.4: Khuynh hng sn xut tiu Barzil [32]
c. Indonesia
Indonesia ng gp c 23.000 tn trong nm 2009 cao hn 12 phn trm
so vi 2009. Tnh hnh sn xut tiu ca Indonexia khng n nh v c xu hng
gim dn qua cc nm. Nm 2000 sn lng tiu t hn 80.000 tn nhng nm
2006 ch t gn 30.000 tn. Nhng Indonesia l mt nc xut khu ht tiu hot
ng trn ton cu 2010 v gi lun thp hn n . Indonesia dn dn ni ln
l nc xut khu tiu ln nht cho th trng M. Theo d liu n thng 7 nm
2009 Indonesia xut khu s lng cao nht 2911 tn trong mt thng nhiu hn
cc con s Vit Nam 1.447 tn v n v 366 tn. [32]
- 15 -
Biu 1.5: Khuynh hng sn xut tiu Indonesia [32]
1.1.5.2 Tnh hnh sn xut tiu Vit Nam
Tiu Vit Nam hin nay c trng ch yu cc tnh ng Nam B, tp
trung nhiu nht Bnh Phc, B Ra Vng Tu, v ng Nai, ch yu trn nn
t . Vng trng tiu tp trung th hai l Ty Nguyn, phn b ch yu hai
tnh ak Lak v Gia Lai.Trong , tiu Ch s Gia Lai c nng sut rt cao, trn
di 4 tn/ha, Mc d vy, din tch trng tiu cc tnh Ty Nguyn cng kh
ln, chim n 13 ngn ha, tnh Bnh Phc vi din tch 12.300 ha, cao nht
nc.
cc tnh Bc Trung B, ni ting nht l vng tiu Qung Tr, c cht
lng tiu cao (thm, cay) v din tch kh tp trung khu vc t Cam L. Cc
tnh khc thuc cc vng trn c din tch trng tiu t hn, v khng mang tnh
cht sn xut hng ha ln, tp trung cung cp nguyn liu cho xut khu. Tiu Ph
Quc ni ting t lu i v cht lng tuyt ho. [16].




- 16 -









Tnh thnh Din tch (ha)
Bnh Phc 12,300
ng Nai 7,586
B Ra-Vng Tu 6,800
ak Nng 5,600
Gia Lai 5,500
Dak Lak 3,500
Bnh Thun 2,300
Qung Tr 2,150
Bnh Dng 1,300
Cc tnh khc 3,069
Tng 50,105
Bng 1.3: Din tch trng tiu
cc tnh thnh Vit Nam. [52]
Biu 1.6: Bn phn b cc tnh trng tiu Vit Nam [52]
http://peppervietnam.com/en/status_pages.php?id=108&id_cat=6
- 17 -
Bng 1.4 Sn lng Vit Nam, th gii giai on 1998- 2010 ( n v: tn) [53]
T trng ca Vit Nam
Nm Vit Nam Th gii
so vi th gii
1998 22.000 206.284 10,7 %
1999 30.000 218.340 13,7 %
2000 36.000 259.186 13,9 %
2001 56.000 308.195 18,2 %
2002 75.000 341.060 22%
2003 85.000 362.160 23,5 %
2004 100.000 323.480 31%
2005 95.000 314.270 30,2 %
2006 100.000 289.230 34,6 %
2007 90.000 271.000 33,2 %
2008 90.000 270.000 34,6 %
2009 100.000 282.900 31%
2010 * 90.000 291.2000 29%

Ghi ch: *: c tnh
http://peppervietnam.com/en/news_detail.php?id=1061&id_groupN=59&id_catN=6
1.1.6 Tnh hnh xut khu tiu ca Vit Nam
Lng ht tiu ca Vit Nam xut khu trong thng 5/2010 l hn 15 nghn
tn, gim 6,4% v kim ngch t 49 triu USD, gim 1,5% so vi thng 4/2010.
Vi kt qu xut khu ca thng 05/2010 nng tng lng ht tiu xut khu ca
nc ta trong 5 thng u nm 2010 ln 60 nghn tn vi kim ngch 184 triu USD.
Lng xut khu nhm hng ny ch tng 11% nhng do n gi xut khu bnh
qun tng 33,4% so vi n gi bnh qun ca 5 thng/2009, mc gn 3100
USD/tn. Do , tr gi xut khu tng ti 48,1% so vi cng k nm 2009. Nh
vy, xut khu ht tiu ca Vit Nam trong 5 thng u nm 2010 tng mnh c v
lng, tr gi v n gi so vi 5 thng/2009.

- 18 -

Biu 1.7: Lng, tr gi, n gi xut khu bnh qun ht tiu ca Vit Nam
nm 2009 v 5 thng u nm 2010
Nm thng u nm 2010, c hai th trng xut khu ht tiu t trn 25
triu USD l Hoa K v c. Trong 5 thng u nm 2010, Hoa K v c tip tc
l hai i tc nhp khu ht tiu ln nht ca Vit Nam. ng u v lng v kim
ngch nhp khu ht tiu ca Vit Nam l th trng Hoa K vi lng t 8,3
nghn tn v tr gi l gn 27 triu USD, tng 91,3% v lng v tng gn 2,1 ln v
tr gi so vi cng k nm trc. ng th 2 l th trng c vi lng t 8,2
nghn tn, tng 91,1% v tr gi l gn 26 triu USD, tng gn 2,5 ln so vi 5 thng
nm 2009.
c bit, lng xut khu ht tiu ca Vit Nam sang hai th trng ny
trong 5 thng u nm chim ti gn 28% trong tng lng xut khu nhm hng
ny ca c nc trong khi con s ny ch l 17% trong 5 thng 2008 v 16% trong
5 thng 2009.
Tnh n ht thng 5/2010, kim ngch xut khu ht tiu ca Vit Nam sang
hu ht cc th trng u t mc tng trng dng so vi cng thi gian mt
nm trc . Ba th trng c kim ngch nhp khu ht tiu ca Vit Nam ln tip
theo u t trn 12 triu USD l n vi 4,4 nghn tn, tng 71,8% v 2,2 ln v
- 19 -
tr gi; Tiu vng quc Rp thng nht vi lng 4,2 nghn tn, gim 12,5%
nhng tr gi li tng 27,4% v H Lan vi 3,7 nghn tn, tng 7,6% v lng v
40,1% v tr gi.
Bng 1.5 Lng, tr gi xut khu ht tiu ca Vit Nam sang mt s th
trng chnh trong 5 thng u nm 2010 so vi cng k 2009.
5 thng u nm 2010
Tc tng/ gim so
vi cng k 2009 (%) Th
hng
Th trng
Lng
(tn)
Tr gi (USD) Lng Tr gi
1 Hoa K 8.304 26.697.566 91.3 108.3
2 c 8.245 25.866.945 91.1 146.9
3 n 4.370 12.042.378 71.8 118.3
4 Rp Thng Nht 4.184 12.071.695 -12.5 27.4
5 H Lan 3.681 12.206.140 7.6 40.1
6 Indonesia 2.305 8.953.978 182.1 288.9
7 Pakistan 2.162 6.160.432 -25.1 8.7
8 Ai cp 1.909 5.267.744 -49.1 -29.5
9 Nga 1.833 5.140.978 -8.3 32.6
10 Ba Lan 1.469 4.086.889 -11.8 33.8
11 Cc th trng khc 21.044 65.808.262 -10.7 19.4
Tng cng 59.506 184.303.066 11.0 48.1

Ngun: Tng Cc Hi Quan
1.1.7 Ch tiu cht lng tiu trng Vit Nam [55]
Cht lng tiu trng Vit Nam c nh gi theo tiu chun Vit Nam
7037:2002. Tiu trng phi c mi thm c trng ca ht tiu trng, cay v khng
c mi v l. Khng c nm mc, cn trng v cc phn xc ca cn trng nhn thy
c bng mt thng (k c knh lp) v phi t cc ch tiu vt l, ha hc,vi
sinh theo tiu chun Vit Nam 7037:2002.
Mc yu cu: Ht tiu SP: Ht tiu s ch
Ht tiu P: Ht tiu sn xut

- 20 -
Bng 1.6 Cc ch tiu vt l ca ht tiu trng
Tn ch tiu Mc Yu Cu
Ht tiu SP Ht tiu P
1. Tp cht l, % khi lng, khng ln hn. 0,5 0,2
2. Ht lp, % khi lng, khng ln hn. 4,0 2,0
3. Ht u inh hoc ht v, % khi lng, khng ln hn. 15 10
4. Khi lng theo th tch, g/l, khng nh hn. 600 600

Bng 1 .7 Cc ch tiu ha hc ca ht tiu trng
Cc ch tiu Mc yu cu

Ht tiu SP
hoc P
Bt ht
tiu
1. m,% khi lng, khng ln hn. 13,0 12,5
2. Tro tng s, % khi lng theo cht kh, khng ln hn. 3,5 3,5
3. Cht chit ete khng bay hi, % khi lng tnh theo cht
kh, khng nh hn.
6,5 6,5
4. Du bay hi, % (ml/100g) tnh theo cht kh, khng nh
hn.
1,0 0,7*
5. Piperin, % khi lng tnh theo cht kh, khng nh hn. 4,0 4,0
6. Tro khng tan trong axit, % khi lng tnh theo cht kh,
khng ln hn.
- 0,3
7. X th, ch s khng ha tan, % khi lng tnh theo cht
kh, khng nh hn.
- 6,5

* Du bay hi cn c xc nh ngay sau khi nghin
- 21 -
Bng 1.8 Cc ch tiu vi sinh vt i vi ht tiu trng ch bin
Tn ch tiu Mc gii hn
1. Coliform, s vi khun trong 1g sn phm 10
2

2. E.coli, s vi khun trong 1g sn phm. 0
3. Salmonella, s khun lc trong 25g sn phm. 0
4. S. aureus, s vi khun trong 1g sn phm. 10
2



Hnh 1.6: Tiu trng [44]
http://binhminhfoods.com/chi-tiet-san-pham/ho-tieu-trang/526/573.html

1.2 Gii thiu v c cht v enzym
Pectin v cellulose l hai thnh phn chnh cu to nn lp v ca tiu. Lp
v ht tiu ny rt kh phn hy bng phng php vt l
1.2.1 Gii thiu chung v pectin:
a. Ngun gc [1], [18], [19]
Pectin l mt polymer ca cc acid polygalacturonic v cc ester methyl ca
chng. Pectin c nhiu qu, c hoc thn cy. Trong thc vt, pectin tn ti di
hai dng: Dng protopectin khng tan, tn ti ch yu thnh t bo di dng kt
hp vi polysaccharide araban, dng ha tan ca pectin tn ti ch yu dch t
bo. Pectin nh mt loi keo gn cht cc t bo thc vt vi nhau, v th ngi ta
gi chng l cht ciment trong cu trc t bo thc vt.
Khi qu cn xanh, ciment l protopectin, protopectin chim t l kh cao,
protopectin khng ha tan trong nc gip qu c cng.
Khi qu chn dn, di tc dng enzyme protopectinase, protopectin s
- 22 -
chuyn sang pectin ha tan (pectin) v araban, lm gim s lin kt gia cc t bo,
qu tr nn mm hn. Trong thc vt cc pectin thng lin kt vi cellulose
vch t bo di dng phc hp.
Hnh 1.7 : Pectin trong cu to ca thnh t bo thc vt [45]
http://www.ebook.edu.vn
Pectin tn ti vi nhng hm lng khc nhau trong qu, c hoc thn ca
mt s loi thc vt: To 10 15%, qu citrus 20 50%, c ci ng 10 20%,
trong cng mt loi qu nhng cc phn khc nhau th hm lng pectin cng
khc nhau.
b. Cu to phn t pectin [18], [19]
Cc cht pectin l cc polysaccharide, mch thng, cu to t s lin kt gia
cc mch ca phn t acid D-galacturonic C
6
H
10
O
7
, lin kt vi nhau bng lin kt
1,4-glucoside. Trong mt s gc acid c cha nhm th methoxyl (-OCH
3
).
Chiu di ca chui acid polygalacturonic c th bin i t vi n v ti hng
trm n v acid galacturonic.





Hnh 1.8 : Cu to ca mt n v pectin [48]
http://files.myopera.com/luutrutailieu/giangday/pectin.doc
- 23 -
Phn t lng ca cc loi pectin tch t cc ngun qu khc nhau thay i
trong gii hn rng ty theo s phn t acid galacturonic v thng thay i trong
phm vi t 10.000 100.000. Trong cc hp cht dng glucid so v chiu di phn
t th pectin cao hn tinh bt nhng thp hn cellulose. V d t ngun to, mn thu
c pectin c phn t lng t 25.000 35.000, trong khi pectin ly t cam li
c phn t lng t ti 50.000.
Tn gi pectin dng ch cc chui polygalacturonic methyl ha 100%.
Tn gi acid pectinic ch cht c methyl ha thp hn 100%. Cn tn gi acid
pectic ch acid polygalacturonic hon ton khng cha nhm methoxyl.
Trong thc tin th tn pectin dng ch c acid pectinic v pectin.
c. Tnh cht ca pectin: [1], [5],[19]
Pectin thuc nhm cc cht lm ng t. Pectin c xem l 1 trong nhng
ph gia thc phm an ton v c chp nhn nhiu nht. c tnh quan trng ca
pectin l khi c mt ca acid v ng n c kh nng to ng (to gel)
HMP (High Methoxyl Pectin): Nhm c ch s methoxyl cao (HMP): MI >
7%, trong phn t pectin c trn 50% cc nhm acid b ester ha (DE > 50%).

Hnh 1.9 : Cng thc HM pectin [45]
http://www.ebook.edu.vn
HMP : To gel bng lin kt hydro
iu kin to gel: [ng] > 50%, pH = 3 - 3,5; [pectin] = 0,5 - 1%.
Cu trc ca gel: ph thuc vo hm lng ng, hm lng acid, hm
lng pectin, loi pectin v nhit .
LMP (Low Methoxyl Pectin): Nhm c ch s methoxyl thp: MI< 7%, t 3
5%, trong phn t pectin c di 50% cc nhm acid b ester ha (DE 50%).
- 24 -


Hnh 1.10: Cng thc LM pectin [45]
http://www.ebook.edu.vn
LMP : To gel bng lin kt vi ion Ca
2+

iu kin to gel: Khi c mt Ca
2+
, ngay c nng < 0,1% min l chiu
di phn t pectin phi t mc nht nh, khi gel c to thnh ngay
c khi khng thm ng v acid.
Cu trc ca gel: Ph thuc vo nng Ca
2+
v ch s methoxyl (MI). Gel
pectin c ch s MI thp thng c tnh n hi ging nh gel agar agar.
d. ng dng
Pectin l cht to gel quan trng nht c s dng to ra cu trc gel
cho thc phm. Kh nng to gel ca n c s dng trong nhng thc phm cn
c s n nh ca nhiu pha. Tc dng to gel ca pectin c s dng ch yu
trong cc sn phm mt tri cy v mt ng, nc qu, ru vang, saPectin
cn c ng dng trong cng ngh dc phm dng ch thuc ung, thuc tim
cm mu trc v sau phu thut. [1], [18], [19]
1.2.2 Cellulose
a. Ngun gc [1], [19],
Cellulose l hp cht cao phn t c cu to t cc lin kt cc mt xch -
D-Glucose, c cng thc cu to l (C
6
H
10
O
5
)
n
hay [C
6
H
7
O
2
(OH)
3
]
n
trong n c
th nm trong khong 5000-14000, l thnh phn ch yu cu to nn vch t bo
thc vt. Trong g l kim, cellulose chim khong 41-49%, trong g l rng n
chim 43-52% th tch.
- 25 -
i vi ngi th cellulose khng c gi tr dinh dng v cellulose khng b
thy phn trong ng tiu ha. Nhng c vai tr trong iu ha tiu ho. ng vt n
c thy phn c cellulose nh enzyme cellulase.
b. Cu to phn t cellulose [1], [19], [35], [41], [49]
Phn t cellulose c cu trc dng mch thng. Cu trc ny to iu kin
hnh thnh cc lin kt hyro gia cc phn t cellulose nm song song vi nhau,
to nn cu trc mng cellulose v vi si (micro fibrin) trong cu trc mng
cellulose ca t bo thc vt.
Cc si ny khng tan trong nc, rt bn v c hc nn to nn lp mng
cellulose bn chc. Thnh phn ha hc ca mng bo v c phn tch, kh
phc tp. Nc chim 60% c cha trong cc khong t do ca mng, 30%
cellulose, cc si cellulose lin kt vi nhau to thnh cc mixen (khong 100 si
cellulose bn li vi nhau to nn mt mixen vi kch thc 5nm, c 20 mixen kt
vi nhau li to nn mt si b (microfibrin) vi kch thc khong 10- 20 nm, v
c 250 si b li to nn si ln (macrofibrin). Cc si an cho vi nhau theo
nhiu hng lm cho mng cellulose rt bn vng, nhng li c kh nng n hi.
gia cc si l khi khng gian cha cc cht v nh hnh gm hemicellulose,
pectin v nc.






Hnh 1.11 Cu to ca phn t cellulose [56]
http://www.scribd.com/doc/38460876/So-An pdf
Cellulose c dng hnh si di, nhiu si kt hp song song vi nhau thnh
chm nh cc lin kt hydro, mi chm (micelle) cha khong 60 phn t cellulose.
- 26 -
Gia cc chm c nhng khong trng, khi ho g khong trng ny cha y
lignin v ta xem lp lignin ny nh l mt lp cement.

Hnh 1.12 S cu to ca si cellulose [50]
http://my.opera.com/thuhien1st/blog/cau
c. Tnh cht celulose [4], [19]
Tnh cht vt l
L cht mu trng, khng mi, khng v. Cellulose khng tan trong nc v
cc dung mi hu c thng thng. Tan trong dung dch Schweitzer, acid v c
mnh nh: HCl, HNO
3
,... mt s dung dch mui: ZnCl
2
, PbCl
2
,Khi un nng vi
H
2
SO
4
, cellulose s b thy phn thnh cc phn t D-glucose cellulose l cht si
hu c hyrat-cacbon phc hp cu thnh nn v cng t bo ca thc vt.
Tnh cht ha hc
Cellulose do cc mt xch -D-Glucose lin kt vi nhau bng lin kt 1-4
glucocid do vy lin kt ny thng khng bn trong cc phn ng thy phn.
Tc dng vi dung dch base
Tc dng vi dung dch acid v c
Tc dng vi dung dch acid hu c
d. ng dng [1], [19]
Cellulose vi tinh th c s dng trong cc loi thc phm calori thp,
trong salad, cc mn trng ming v kem. Kh nng hydrat ha v phn tn ca
- 27 -
chng c ci thin r rt khi s dng chung vi mt lng nh carboxymethyl
cellulose.
1.2.3 Gii thiu v enzym
Enzyme l cht xc tc sinh hc c thnh phn c bn l protein. Trong cuc
sng sinh vt xy ra rt nhiu phn ng ha hc, vi mt hiu sut rt cao, mc d
iu kin bnh thng v nhit , p sut, pH. S d nh vy v n c s hin din
ca cht xc tc sinh hc c gi chung l enzyme. Nh vy, enzyme l cc
protein xc tc cc phn ng ha hc.
Trong cc phn ng ny, cc phn t lc bt u ca qu trnh c gi l c
cht (substrate), enzyme s bin i chng thnh cc phn t khc nhau. Tt c cc
qu trnh trong t bo u cn enzym. Enzym c tnh chn lc rt cao i vi c
cht ca n. Hu ht phn ng c xc tc bi enzym u c tc cao hn nhiu
so vi khi khng c xc tc. C trn 4.000 phn ng sinh ha c xc tc bi
enzym.
Hot tnh ca enzym chu tc ng bi nhiu yu t. Cht c ch l cc phn
t lm gim hot tnh ca enzym, trong khi yu t hot ha l nhng phn t lm
tng hot tnh ca enzym [4], [6], [19], [43].
1.2.3.1. Tnh cht ca enzym
Enzym c bn cht l protein nn c tt c thuc tnh l ha ca protein. a
s enzym c dng hnh cu v khng i qua mng bn thm do c kch thc
ln.
Tan trong nc v cc dung mi hu c phn cc khc, khng tan trong ete
v cc dung mi khng phn cc.
Khng bn di tc dng ca nhit , nhit cao th enzym b bin tnh.
Mt trng axt hay baz cng lm enzym mt kh nng hot ng.
Enzym c tnh lng tnh: Ty pH ca mi trng m tn ti cc dng:
cation, anion hay trung ha in.
- 28 -
Enzym chia lm hai nhm: Enzym mt cu t (ch cha protein) nh pepsin,
amylase... v cc enzym hai cu t (trong phn t cn c nhm khng phi protein)
[5], [19]
1.2.3.2. Cc yu t nh hng n kh nng hot ng ca enzym
Cc hot ng enzyme b nh hng bi nhiu yu t, nh nhit , pH,
nng enzyme, nng ca c cht v cht c ch. L yu t trc tip tc ng
nhiu n hot ng ca enzym.
a. nh hng ca nhit
Vn tc phn ng do enzym xc tc ch tng theo nhit trong mt gii hn
xc nh m phn t enzym cha b bin tnh. Nhit ng vi hot enzym
cao nht gi l nhit ti u ca enzym. a s enzym c nhit ti u khong
40-50
o
C. Tuy nhin nhit ti u ca mt enzym khng c nh m c th thay
i ty c cht, pH mi trng, thi gian phn ng.Nhit m enzym b mt
hon ton hot tnh xc tc gi l nhit ti hn vo khong trn 70
0
C. nhit
ti hn enzym b bin tnh, t khi c kh nng phc hi li c hot . Ngc li
nhit di 0
0
C hot enzym tuy b gim nhng li c th tng ln khi a v
nhit bnh thng [19].
Hnh 1.13 nh hng nhit n tc phn ng [43]
b. pH mi trng
pH mi trng nh hng r rt n phn ng enzym v n nh hng n
mc ion ha c cht, enzym v nh hng n bn protein enzym. a s
- 29 -
enzym bn trong gii hn pH gia 5-9, bn ca enzym c th tng ln khi c c
cht, coenzym, Ca
2+

pH lm thay i trng thi ion ha cc nhm nh chc trung tm hot
ng enzyme lm thay i kh nng phn ng ca cc nhm ny trong phn ng
xc tc v c th lm thay i cu trc trung tm hot ng ca enzyme.
pH cng lm thay i trng thi ion ha ca c cht, ti pH ti thch phn t
c cht c ion ha ti trng thi thch hp nht cho s kt hp vi enzyme [19].


Hnh 1.14: nh hng pH n tc phn ng [43]
c. nh hng ca nng enzyme v c cht
T l phn ng ca enzym xc tc ph thuc vo nng ca enzym v c
cht. Nh vy t l phn ng enzym tng khi tng nng enzym v cht nn.







Hnh 1.15: nh hng ca nng enzym Hnh 1.16: nh hng ca c cht
ln tc phn ng [43] ln tc phn ng [43]
T

p
h

n
g

Nng enzym
:im bo ha
T

p
h

n
g

Nng enzym
- 30 -
Gi nguyn nng enzym, tng tc phn ng bng cch tng nng
c cht ln n mt im no , tc phn ng ca enzym tng nhanh sau
chm li v khng tng na. iu ny l do hot ng ca cc phn t enzym ti
thi im gn nh bo ha vi c cht. Nh vy nng cht nn l cao, nhit
v pH gi khng i, t l phn ng t l thun vi nng enzym [43].
d. nh hng ca cc cht km hm.
Hot ca enzym c th b thay i di tc dng ca mt s cht c bn
cht ha hc khc nhau. Cc cht lm gim hot ca enzym nhng khng b
chuyn ha bi enzym c gi l cht km hm hoc cht c ch [19].
Cht km hm c c cu trc tng t vi c cht, do c kh nng kt hp
vo trung tm hot ng ca enzym chim ch kt hp vi c cht gi l cht km
hm cnh tranh.
Cht km hm c kh nng kt hp vi enzym ch khc vi trung tm hot
ng lm thay i cu trc khng gian ca phn t enzym theo hng khng c li
cho hot xc tc ca n lm gim tc phn ng xc tc gi l cht km hm
khng cnh tranh. [19]
1.2.4 Gii thiu enzyme pectinase
Enzyme pectinase l enzyme xc tc s phn hy ca cc polymer pectin. S
phn hy pectin trong t nhin thng xy ra khi tri cy chn. Nhng enzyme ny
v vy c mt vai tr ht sc quan trng trong qu trnh bo qun tri cy v rau
qu. [5]
Enzyme pectinase cng c ng dng nhiu trong qu trnh ch bin thc
phm, c bit l kh nng lm trong nc qu qua kh nng kim sot c
nht ca sn phm [1].
1.2.4.1 Phn loi enzyme pectinase [1],[4], [5], [6], [7]
Enzyme pectinase c th c phn loi theo c ch tc dng ca chng
Pectinesterase (PE): Xc tc s thy phn ca cc nhm methyl ester. To
thnh acid pectinic hoc acid pectic v methanol.
- 31 -
Pectinesterase thu c t cc ngun khc nhau c gi tr pH ti u khc
nhau. Nu thu t ngun vi sinh vt (nm mc, x khun, vi khun) th pH
opt
=
4,5-5,5, ngun thc vt th c pH
opt
= 5,0-8,5. Pectinesterase t nm mc c T
opt
=
30-40
o
C v b v hot 55-62
o
C. Pectinesterase thng c hot ho bi cc ion
Ca
2+
v Mg
2+
.
Polygalacturonase cn c tn gi l poly 1,4-galacturoniglucanohydrolase,
xc tc s phn ct cc mi lin kt 1,4-glycosid. Polygalacturonase t gp trong
thc vt nhng c ch yu mt s nm mc v vi khun. Polygalacturonase l
mt phc h enzyme gm c nhiu cu t v thng c tnh c hiu cao i vi c
cht. Trn c s tnh c hiu v c ch tc dng vi c cht, enzym
polygalacturonase c chia thnh:
Polymethylgalacturonasec (1,4-galacturonite-methylesglucanohydrolase)
tc dng trn polygalactorunic acid c methoxyl ho (tc l pectin). Enzyme
ny li c phn thnh hai nhm nh l endo-glucosidase-polymethyl
galacturonase kiu I v exo-glucosidase-polymethylgalaturonase kiu III.
Polygalacturonase enzyme tc dng trn pectic acid hoc pectinic, cng
c chia thnh hai nhm nh l endo-glucosidase-polygalacturonase kiu II v
exo-glucosidase-polygalacturonase kiu IV. Enzyme endo-glucosidase-Polymethyl-
galacturonase kiu I l enzyme polymethylgalacturonase dch ho pectin c mc
methyl ho cng cao th b thy phn bi enzyme ny cng nhanh v cng c hiu
qu. Trong dung dch, khi c mt ca enzyme pectinesterase th hot ca enzyme
ny thng b gim. Enzyme ny rt ph bin trong cc vi sinh vt, c bit l nm
mc Aspergillus niger, Aapergillus awamori.
Pectate lyase (PEL): Xc tc s phn ct cc n v galacturonate khng b
ester ha. C hai enzyme exo-PEL (exo-Poly (1,4-D-galacturonide) lyase) v endo-
PEL (endo-poly(1,4-D-galacturonic) lyase) u tn ti. Pectate v pectin c lng
methoxyl thp l cc c cht thch hp hn c cho cc enzyme ny.
- 32 -
Ni chung, c hai enzyme ny u c khong pH ti a nm trong khong t
8,0-11, u cn ion Ca
2+
hot ng. Pectate lyase khng c tm thy trong cy
xanh, nhng c vi khun v nm.
Cc enzyme vi sinh vt ngoi bo ny ng mt vai tr rt quan trng trong
qu trnh gy bnh thc vt, gy ra s phn hy m ca thnh t bo, lm mm v
lm mc m thc vt.
Ngoi ra cn c:
Pectin-transeliminase hay cn c gi l poly1,4-galaturonite
methylesteglucanoliase, l enyme tc dng trn pectin v pectinic acid.
Polygalactorunate-transeliminase cn c gi l poly -1,4 D-galaturonite
glucanoliase, l enzyme tc dng trn pectic acid v pectinic acid.
Pectin lyase (PNL): Xc tc s phn ct cc n v galacturonate b ester
ho. Tt c cc PNL u l endo-enzyme.
1.2.4.2 Vi sinh vt sn xut enzym pectinase
Pectinase c sn xut ch yu bi nm mc Aspergillus sp., Botrytis
cinerea, Fusarium moniliforme, Rhizopus stolonifer, Trichoderma sp., Neurospora
crassa [4]
Nm men: Saccharomyces fragillis
Vi khun: Bacillus polymyxa, Flavobacterium pentinivorum, Klebsiella
aerogenes [5], [20]
1.2.4.3 ng dng pectinase
Pectinase thng c s dng trong cc ngnh cng nghip thc phm nh:
Sn xut ru vang, nc qu v nc ung khng c cn, cc mt hng t qu:
nc qu c c, mt nh, mt ng, sn xut nc gii kht, sn xut c ph v c
ph ha tan [4], [19].
S dng ch phm pectinase trong trch ly cc dc liu ng y. ng dng
pectinaza trong chn nui b sung vo khu phn n ca tru b, ngng g


- 33 -
1.2.5 Gii thiu v enzym cellulase
Enzym cellulase l mt phc h enzym c tc dng thu phn cellulose
thng qua vic thu phn lin kt 1,4--glucosid trong cellulose to ra sn phm
glucose cung cp cho cng nghip ln men. [2]
Cellulase dng ch mt loi enzyme c sn xut ch yu l do nm , vi
khun , v cc vi sinh vt n bo, thc vt v ng vt. [35]
1.2.5.1 Phn loi enzyme cellulase
Cellulase c phn thnh 5 loi da vo phn ng xc tc. l Endo-
cellulase, Exo-cellulase 2-4, Cellobiase, Cellulose phosphorylases, oxidative
cellulase.[35]
Endo-cellulase b gy cc lin kt bn trong lm ph v cu trc mng tinh
th ca cellulose v to ra cc si polysaccharit ring l.
Exo-cellulase ct 2 4 n v t u cc chui m c ct bi endo-
cellulase, kt qu to thnh cc tetrasaccharides hoc disaccharide.
Cellobiase hay beta-glucosidase thy phn exo-cellulase to thnh cc
monosaccharit ring r.
Oxidative cellulose thc hin phn ng trng hp cellulose bng cch thc
y cc phn ng..
Cellulose phosphorylase thc hin phn ng trng hp cellulose bng cch
dng cc phosphate thay v s dng nc.
S minh ha c ch tc dng ca enzym cellulase [1].
Cellulose Cellulose Cellobiose Glucose


Cellulase C
1
Cellulase C
x
-glucozidase

1.2.5.2 Vi sinh vt tng hp enzym cellulase.
T nm si: mt s loi nm si c kh nng sinh ra mt lng ln cellulase,
Altenaria tenuis, Aspergillus fumigatus, Aspergillus flavus, Celluvibrio gilvus,
Fusarium culmorum, Trichoderma koningi...Trong Trichoderma v Aspergillus
- 34 -
c nhiu nh khoa hc nghin cu sn xut cellulase. [2], [19], [24], [35],
[37]
T x khun: Actinomyces celluloseac, Actinomyces diastatitus, Actinomyces
roseus, Actinomyces griseus, Actinomyces melamocylas...[1], [7]
T vi khun: Bacillus megaterium, Mensenteroides, Pseudomonas
fluorenscons, Clostridium sp, Acetobacter xylinum...[7]
1.2.5.3 ng dng cellulase
ng dng trc tin ca cellulase i vi ch bin thc phm l dng n
tng hp thu, nng cao phm cht v v v lm mm nhiu loi thc phm thc
vt. c bit l i vi thc n cho tr con v ni chung cht lng thc phm c
tng ln.
Mt s nc dng cellulase x l cc loi rau qu nh bp ci, hnh,
c rt, khoai ty, to v lng thc nh go. Ngi ta cn x l c ch, cc loi to
bin
Ngoi ra cn s dng cellulase trong sn xut agar-agar, hay phc h citase
trong c cellulase trong sn xut bia. t bit l vic s dng ch phm cellulase
tn thu cc ph liu thc vt em thy phn, dng lm thc n gia sc v cng
ngh ln men [1].
Cellulase c ng dng ci thin gi tr dinh dng ca thc n gia sc,
gia cm, trch ly cc cht t thc vt, t cy thuc, ng ha cc ph liu giu
cellulose sn xut ethanol [5]. [6]
1.3 Gii thiu ch phm Biopep-21, Biopep-22, Biopep-31, Biopep-32.
Mc ch ca nghin cu ti ny l bc v ht tiu en thnh tiu trng.
Hai thnh phn chnh ca v ht tiu l pectin v cellulose. Chng rt kh tch ra
bng phng php vt l. V d tiu c ngm 10-15 ngy v mm v c ch
xt bng my v c km theo s dng ha cht. Vic s dng ch phm sinh hc
nhm mc ch phn gii pectin v cellulose bc v ht tiu trong thi gian ngn,
nng cao cht lng ht tiu. Ch phm Biopep-21 c sn xut t Aspergillus
- 35 -
niger , Biopep-22 t Aspergillus oryzae, Biopep-31 t Bacillus polyfermenticus,
Biopep-32 t Bacillus amyloliquefaciens
1.3.1 Ch phm Biopep-21
c sn xut t chng Aspergillus niger. Chng vi sinh vt ny c cung
cp t phng th nghim Khoa Cng ngh Sinh hc, trng i hc Quc t.
Apergillus niger thng thy trn lng thc bo qun, lc va mi phn lp
thng c u nht nhy. Tuy nhin, tip tc nui cy, tnh cht ny bin mt
Apergillus niger trn thch Czapek 25
0
C, ng knh t 4-5 cm trong 7 ngy,
dng thm th hoc dng si, mu en, mu nu en hoc nu tm, pht trin rt
nhanh, thi k u si mc dng lng d trng.
u ht nh: To, hnh cu, ng knh 300-500 m, thm ch ti 1000 m,
mu ging nh mu ca khm mc, khi mi th khng mu.
Cung ht nh rt di t 200 m n vi nm, mt bng nhn, khng mu
hoc pha trn cung mu vng nht.
Bng: Hnh cu, khng mu hoc vy mu nu vng, ng knh 50-100m.
Cung: Hai hng st bng l cung th bnh, trong ti nu thng c vch
ngn 15-25 x 4,5-6,0 m, th bnh 7,0-9,5x3-4m.
Ht nh: Hnh cu ti tn cu 3,5x5m, mt x x, mu nu en, khng u,
mn cm, gai v ln gn.
Hch nm: Thng thng xut hin, hnh thi t khng nh n hnh cu,
mu xm hoc mu en. [17]

Hnh 1.17 : Aspergillus-niger [54]
http://permimalang.wordpress.com/2007/12/12/aspergillus-niger/
- 36 -
C nhiu loi enzym c sc phn gii mnh nh amylaza, lypaza, proteaza,
pectinnaza, cellulazec th phn gii cht hu c sinh ra nhiu axit hu c. [4],
[5], [17], [20], [36]
Nh vy trong qu trnh ln men s dng ch phm Biopep-21, pectin v
cellulose trong v ht tiu s b phn gii lm bc c v ht tiu.
1.3.2 Ch phm Biopep-22
c sn xut t chng Aspergillus oryzae . Chng vi sinh vt ny c
cung cp t phng th nghim Cng ngh Sinh hc, trng i hc Quc t.
Aspergyllus oryzae l mt loi vi nm thuc b Plectascales, lp
Ascomycetes. C th sinh trng ca n l mt h si bao gm nhng si rt mnh,
chiu ngang 5-7 m, phn nhnh rt nhiu v c vch ngn, chia si thnh nhiu
bao t bo (nm a bo).
T nhng si nm ngang ny hnh thnh nhng si ng thng gi l cung
nh bo t, c c quan sinh sn v tnh.
Cung nh bo t ca Aspergyllus oryzae thng di 1-2 mm nn c th
nhn thy bng mt thng. Pha u cung nh bo t phng ln gi l
bng.
T bng ny phn chia thnh nhng t bo nh, thun, di, gi l nhng t
bo hnh chai.
u cc t bo hnh chai phn chia thnh nhng bo t nh vo nhau, nn
gi l nh bo t.
nh bo t ca Aspergillus oryzae c mu vng lc hay mu vng hoa
cau
c im ca ging Aspergyllus oryzae l giu cc enzyme thy phn ni
bo v ngoi bo (amylase, protease, pectinase, cellulase), ta rt hay gp chng
cc kho nguyn liu, trong cc thng cha ng bt, go ht nhng khng
c ra sch, cn b bia, b ru, li ng, b sn Chng mc v pht trin
c khi thnh lp mc, c mu sc en ,vng Mu do cc bo t gi c mu sc.
- 37 -
Cc bo t ny, d b gi cun b ay xa v ri vo u khi gp iu kin thun li s
mc thnh mc mi.


Hnh 1.18: Aspergillus oryzae [51]
http://www.nrib.go.jp/English/annai/introduce/microorganism.htm
Nh vy trong qu trnh ln men s dng ch phm Biopep-22, pectin v
cellulose trong v ht tiu s b phn gii lm bc c v ht tiu [4], [17].
1.3.3 Ch phm Biopep-31
c sn xut t chng Bacillus polyfermenticus l vi khun c li, c c
lp ln u tin t khng kh bi Tin s Terakado nm 1993.
Bacillus polyfermenticus c ng dng lm ch phm sinh hc (v d Biotech-
Shrimp) phn hy b mn hu c v cc cht thi [1], [20], [22]
1.3.4 Ch phm Biopep-32
c sn xut t chng Bacillus amyloliquefaciens l mt loi vi khun hiu
kh, gram dng c pht hin trong t nm 1943 bi nh khoa hc Nht Bn l
Fukumoto. Bacillus amyloliquefaciens giu enzym thy phn ngoi bo (amylase,
protease). Enzym t Bacillus amyloliquefaciens s dng cho nhiu mc ch
cng nghip nh sn xut giy, vi, bt git sinh hc, v t bit lm ch phm sinh
hc (v d BIOnAQUA1-MV) x l nc thi, b mn hu c. [1], [20], [28], [31],
[35], [38].


- 38 -
1.4 Gii thiu piperine [10], [27], [33]
Piperine l mt ankaloid c v rt cay l thnh phn chnh to nn v cay ca
cy h tiu (pha long 200.000 ln vn cn cay). Piperine c c lp v xc nh
cu trc t ht tiu en t nm 1820. Ngy nay ngi ta tng hp c piperine
t allylic. Piperine tng hp c mu xanh l cy trong khi piperine c lp t thin
nhin c mu vng nht.
Tn IUPAC ca piperine :
1-[(2E,4E)-5-(1,3-benzodioxol-5-yl)penta-2,4-dienoyl]piperidine
Cn c tn khc l :
5-(3,4-methylenedioxyphenyl)-2,4-pentadienoyl-2-piperidine
piperoylpiperidine
Cng thc phn t ca piperine: C
17
H
19
NO
3


Hnh 1.19. Cng thc phn t ca piperine: C
17
H
19
NO
3
Khi lng phn t: 285,338 g/mol
T trng: 1,193 g/cm
3

Nhit nng chy: 128-130
o
C, 403
o
K, 266
o
F
Piperine c tc dng chng trm cm, chng oxy ho, tc dng lm gim
huyt p [40]. Piperine cn c tc dng lm tng hiu lc sinh hc ca thuc, kch
thch h thng tun hon v tiu ha.[16]


- 39 -
1.5 Gii thiu aflatoxin v ochratoxin A
1.5.1 Aflatoxin
Trong qu trnh sinh trng v pht trin, vi sinh vt c kh nng to ra
nhng cht ha hc c kh nng gy c cho sinh vt khc. Cc cht ny c gi
l c t.
Nm mc thuc nhm vi sinh vt d dng. Chng ch c kh nng nhn cht
dinh dng dng ha tan. Trong qu trnh trao i cht xy ra t bo chng li
c kh nng chuyn ha cc cht ha tan thnh cc cht khng ha tan nh
lignocelluloseNgoi ra, chng cn c th to ra cc cht c. Cc cht c ca
nm mc c gi chung l c t vi nm (mycotoxins). Cc c t ny thng gy
ng c mnh v c kh nng gy ung th cho ngi v ng vt. [8]
Aflatoxin (vit tt ca Aspergillusflavus toxins ) l mt mycotoxin c tit
ra t nm Aspergillus v Aspergillus parasiyicus cc chng nm ny pht trin
mnh cc loi thc phm nng sn, thc n gia sc, gia cm nh ng, lc, u...
trong cc iu kin thun li nng v m, nht l iu kin thi tit Vit Nam.
i vi ngi, aflatoxin gy ung th gan v gy cht ngi.[30]
Aflatoxin c chia ra lm nhiu loi khc nhau. Trong aflatoxin B
1
l
loi c t c c tnh mnh nht.[8]

Hnh 1.20 Cu trc ha hc ca aflatoxin B
1
[55]
http://www.probioticonline.net/


- 40 -
1.5.2 Ochratoxin A
Ochratoxin A l mt nephrotoxin, ln u tin tm thy nm mc
Aspergillus ochraceus vng Nam Phi [8]. Ochratoxin c kh nng gy ung th cho
con ngi, gy t bin v tn thng DNA do phn ng oxi ha. [39]

Hnh 1.21: Cu trc ha hc ca ochratoxin A [46]
http://www.fao.org/docrep/t1838e/t1838e0a.htm




- 41 -
CHNG 2: NGUYN LIU V PHNG PHP NGHIN CU
2.1 Nguyn liu v phng php
2.1.1 Nguyn liu
Aspergillus niger , Aspergillus oryzae, Bacillus polyfermenticus, Bacillus
amyloliquefaciens c cung cp bi phng th nghim Khoa Cng Ngh
Sinh Hc, Trng i Hc Quc T.
Tiu en mua ti nh vn. Khu 10, Th Trn Tn Ph, Huyn Tn Ph,
Tnh ng Nai.
Malt
Pectin
Tru
Cm go
Cn 70%, 96%
Ammonium sunlfate
Dung dch NaOH, HCl iu chnh pH
Nc ct
Dung dch ZnSO
4
5%
Dung dch DNS (3,5- Dinitro Salicylic)
Antron tinh khit: Ha tan 2g Anthron trong 1 lt axt sulfuric m c v gi
trong t lnh. Thuc th ny khng th c s dng sau hai ngy.
CMC (Carboxymethyl cellulose )
Glucose 0,01% (0,01g glucose ha tan 100ml nc)
2.1.2 Thit b
Giy lc
B iu nhit
a thy tinh
ng nghim
R nha
- 42 -
Khn
Vi lc
My o pH
My o Brix
My xay sinh t
Bp in
Cc thy tinh
Erlen
Nhit k
Thit b ct nc.
Cn
My quang ph
2.2 Phng php nghin cu
Tt c cc th nghim c thc hin ti phng th nghim Khoa Cng Ngh
Ha Thc Phm, Trng i Hc Lc Hng.
2.2.1 Phng php vt l
2.2.1.1 Kim tra m ht tiu en
Cho 10g ht tiu en c cho vo chn v c t vo t sy, sy nhit
105
0
C cho n khi trng lng ht tiu khng i.

Cho 10g ht tiu en c t vo cc sau cho thm nc vo mt lng
nh nhau. Tiu c ngm vi nhiu thi lng nh nhau 45 pht, 90, 135, 180
ng vi mi thi gian ngm, ht tiu loi b nc v c cn, ghi nhn gi tr
trng lng.
Ngm ht tiu en cho n khi ht tiu bo ha.
m ht tiu en c tnh theo cng thc sau:
%W= (m
2
m
1
)/ m
1
x 100
Trong : %W m ht tiu (%)
m
1
: Trng lng ban u ca ht tiu (g)
m
2
: Trng lng ht tiu sau khi ngm (g)
- 43 -
2.2.1.2 Loi b v ht tiu
Ht tiu sau khi ln men c t vo thau nc ngm trong 5 pht sau
ch xt bc ht lp v ca ht tiu cho n khi khng th bc v hn c na.
Ra ht tiu bng nc sch nhiu ln ht tiu sch. Sau ht tiu c t vo
t sy nhit 55-60
0
C ht tiu t m 12-13%.
2.2.1.3. Phn loi ht tiu sau ln men
Ht tiu sau khi ln men c ra sch v sy t m 12-13%, ht tiu
c phn thnh hai cp tiu bc v v cha bc v. Sau , ht tiu mi cp
c tnh v tnh theo cng thc sau
% P = LP / TP x 100
Trong : % P: T l phn trm ht tiu mi cp (%)
LP: S lng ht tiu mi cp
TP: Tng s ht tiu trong hai cp [29]
2.2.2 Phng php c hc
2.2.2.1. Kim tra cht rn ha tan chit xut t ht tiu
Ht tiu c nghin trong 5 pht bng my xay sinh t, v sau cn
10g ht tiu bt cho vo Erlen 250ml. Thm 110ml nc ct vo Erlen v lc trong
1 pht trch xut cc cht rn ha tan t tiu bt. Ly dung dch em lc qua giy
lc v cho 1 git dung dch lc ca ht tiu c lc a vo l trn khc x k
kim tra cht rn ha tan t ht tiu. [29]
2.2.2.2. Kim tra pH ca dung dch ht tiu
Ht tiu c nghin trong 5 pht bng my xay sinh t, v sau cn
10g ht tiu bt cho vo Erlen 250ml. Thm 110ml nc ct vo Erlen v lc trong
1 pht trch xut cc cht rn ha tan t tiu bt. Ly dung dch em lc qua giy
lc. pH ca dung dch lc ca ht tiu c o bng my o pH. [29]
2.2.2.3 Kim tra nhit ca ht tiu trong qu trnh ln men.
Cho 50g ht tiu ht tiu c ngm vi thi gian ph hp c c
m ti u. Khi ht tiu t m ti u, c chuyn giao cho vi t v ch cho
n khi cc nc d tha c loi b, v sau tt c chng c t vo gi
- 44 -
nha. B sung t l ch phm thch hp v trn u hn hp vi ht tiu. Nhit k
c t vo khi ln men, hng ngy nhit ln men khi c kim tra.
2.2.3 Phng php ha sinh
2.2.3.1 Chun b mi trng malt cho vi sinh vt [9]
Cn 200g malt, xay nhuyn malt bng my sinh t. Cho malt xay vo
erlen cha 1 lt nc. Dng a thy tinh khuy u v t vo b iu nhit gi
52
0
C trong 20 pht, sau 63
0
C trong 30 pht, 73
0
C trong 30 pht, v cui cng
100
0
C trong 15 pht [9]. Tinh bt c thy phn bi enzym amylase to mi
trng thch hp cho s pht trin vi sinh vt. Dung dch malt c lc bng giy
lc.
Kim tra nng ng sucrose, nu gi tr ny cao hn 10
0
Brix th tin
hnh pha long n gi tr 10
0
Brix. Nng ng sucrose thp hn 10
0
Brix, tin
hnh b sung ng glucose t n 10
0
Brix.
Kim tra pH ca dung dch malt phi cao hn 4,5. Nu gi tr pH thp hn
phi iu chnh pH ca dch malt bng dung dch NaOH v ngc li.
Chun b mi trng thch nui cy, b sung agar vo dch malt 16-18g/l.
Sau un si dch malt trong 5 pht ha tan hon ton agar. Tin hnh ng
nghim (1/4 ng nghim) v a petri. ng nghim c nt cht bng bng khng
thm nc v qun giy nhm, a petri c gi giy kn. Chng c t vo ti
nha chu nhit v t vo thit b tit trng 121
0
C trong 15 pht. Sau ly ng
nghim t nghing v i mi trng ng li. Sau hai ngy nu mi trng khng
b nhim tin hnh nui cy vi sinh vt: Aspergillus niger , Aspergillus oryzae ,
Bacillus polyfermenticus, Bacillus amyloliquefaciens.
2.2.3.2 Phng php xc nh hot tnh ca enzym [11]
1. Xc nh hot tnh ca enzym cellulase bng phng php so mu.
a) Nguyn tc:
Phng php ny da trn s thy gii CMC (sodium carboxymetyl cellulose)
bng enzym pH = 5,0 v 40
0
C. Sau phn ng thy phn s to ra mt lng
- 45 -
ng kh, ng kh s phn ng vi axit 3,5- dinitrosalicylic v c xc nh
bng my o mt quang bc sng 540nm
b) Cch thc hin:
Phn ng enzym:
Ht 1ml dung dch mu vo ng nghim (18x130mm) v n nh
nhit 40 0.1
0
C trong 5 pht. Cng dung dch c cht n nh nhit
ny trong 5 pht.
Thm 1ml dung dch c cht vo ng nghim ng dung dch mu v lc
u.
phn ng xy ra trong khong chnh xc 40 0,1
0
C, 10 pht.
Thm vo 4ml dung dch DNS, lc u tht k ngng phn ng enzym .
trnh s bc hi nc, dng mt ming nilon hay parafilm dn ly
ming ng nghim v t ng nghim vo nc si trong 15 pht.
a ng nghim v nhit phng bng cch t ng nghim vo mt ni
cha nc lnh.
o hp thu OD 540 nm. Dng nc ct lm i chng. (A
T
)
Phn ng th khng:
Ht 1ml dung dch mu vo ng nghim (18x130mm) v n nh nhit
nhit 40 0.1
0
C. Cng dung dch c cht n nh nhit ny
trong 5 pht.
Thm vo 4ml dung dch DNS. Lc u.
Thm 1ml dung dch c cht vo ng nghim ng dung dch mu v lc
u.
trnh s bc hi nc, dng mt ming nilon hay parafilm dn ly
ming ng nghim v t ng nghim vo nc si trong 15 pht.
a ng nghim v nhit phng bng cch t ng nghim vo mt ni
cha nc lnh.
o hp thu OD 540 nm. Dng nc ct lm i chng. (A
B
)
Phn ng ca cc dung dch ng glucose chun v nc ct:
- 46 -
Ht ln lt 1ml cc dung dch ng glucose chun vo cc ng nghim
(18 x 130mm).
Thm vo 1ml dung dch c cht v lc u.
Thm vo 4ml dung dch DNS. Lc u.
Dn nilon hay parafilm ln ming ng nghim, t vo ni nc si trong
15 pht.
a v nhit phng bng ni nc lnh .
o hp th OD 540 nm. S dng nc ct lm i chng (A
G
)
i vi nc ct: Lm tng t nh trn. Thay dung dch glucose chun
bng nc ct. o OD 540 nm. (A
W
)
nh ngha:
Mt n v hot tnh CMCase c nh ngha l s lng enzym cn thit
gii phng ra ng kh (glucose) tc mol/pht di cc iu kin thc
nghim.
c) Tnh ton:
Tnh h s glucose:
Hiu chun hp thu OD ca dung dch ng glucose chun bng cch:
A
G
-A
W
.
i vi mi nng pha long dng ng biu din s bin thin gia
OD v nng glucose (mg/ml). ng ny phi l mt ng thng i qua
gc ta .
Chia nng (mg/ml) cho hp thu OD ca mi dung dch ng chun
thu c gi tr h s glucose (mg/ml), tnh gi tr h s trung bnh.
H s glucose c tnh bng cng thc:



Trong :
F: H s glucose.
F=
A
G
- A
W
C
G
(mg/ml)
- 47 -
C
G
: Nng ca dung dch glucose chun (mg/ml).
A
G
: hp thu OD ca dung dch glucose chun.
A
W
: hp thu OD ca phn ng vi nc ct.
Xc nh hot tnh enzym:
S dng cng thc sau:
CMCase(u/g)=(A
T
-A
B
)*F* * * *

Trong :
A
T
: hp thu OD ca dung dch phn ng enzym.
A
B
: hp thu OD ca phn ng th khng.
F: H s glucose (mg/ml).
1000: Chuyn t mg sang g.
180 : Trng lng phn t ca glucose i t g sang mol.
10 pht: thi gian phn ng.
C: Nng dung dch mu (g/ml).
2. Xc nh hot tnh ca enzym pectinase bng phng php so mu
L phng php phn tch da trn vic so snh cng mu ca dung
dch nghin cu vi cng mu ca dung dch tiu chun c nng xc nh.
Dng phng php so mu ch yu xc nh lng nh cc cht, phn tch bng
phng php ny mt t thi gian hn so vi cc phng php ha hc khc.
a) Nguyn tc:
Xc nh lng galacturonic axit l sn phm ca qu trnh thy phn pectin
di tc dng ca pectinase khng kt ta bng ZnSO
4
.
b) n v hot tnh:
Mt n v hot tnh pectinase l lng enzym trong iu kin tiu chun (nhit
30 pht, pH= 3,9 4,1, thi gian phn ng l 60 pht vi nng pectin trong
mi trng phn ng l 0,66%, mc thy phn pectin l 30%) c kh nng xc
tc, lm bin 1g pectin sau 60 pht thnh axit galacturonic.
c) Ch khi dng phng php so mu:
- 48 -
Mu ca cht nghin cu phi chn lc, mi cht cho mu tng t hoc lm
gim cng mu ca cht nghin cu, cn phi c tch ra hoc lin kt vo
nhng phc khng mu.
Mu khng c thay i trong sut qu trnh o mu.
Mu khng c thay i r rt theo axit ca dung dch v lng thuc th
thm vo.
Mu khng c thay i theo nhit , nu c s thay i th cn phi o
nhit xc nh.
Cng mu phi t l vi nng ca cht nghin cu.
Nng ca dung dch em so mu khng c qu thp hay qu cao v mt ta
kh phn bit s thay i cng mu ca nhng dung dch nh th.
d) Tin hnh:
Cho 20ml dung dch pectin 1% vo ng nghim ri tip theo cho 10ml dung
dch emzym 1%, lc u cho vo t m 30
0
C (dung dch pectin c pH 3,9-4,1).
Sau 60 pht ly ra thm vo 2ml ZnSO
4
15%, em lc qua giy lc, dung dch lc
c em pha long gp 5 ln dng cho phn ng vi Antron.
Ly 5ml Antron cho vo ng nghim, ri cho 2,5 ml dung dch lc pha long,
lc mnh trong thi gian 10 pht. Sau t trong ni cch thy 70
0
C trong thi
gian 12 pht.
Sau ly ra lm ngui n nhit phng v tin hnh o gi tr OD bc
sng 584 nm.
e) Tnh kt qu:
Hot tnh pectinase c tnh bng cng thc:

vht/g MT=

Trong :
OD: Mt quang ca dung dch sau khi phn ng vi antron.
- 49 -
m: Lng enzym ly tin hnh phn ng (g hoc ml), phng php so mu cho
php sai s 5%.
2.2.4 Phng php vi sinh
2.2.4.1 Phng php cy chuyn v nhn ging vi sinh vt. [9]
c th nui cy vi sinh vt cn phi thng xuyn cy vi sinh vt t mi
trng ny sang mi trng khc v phi m bo trong khi cy chuyn khng lm
nhim vi sinh vt ang s dng vi nhng vi sinh vt khng mong mun t mi
trng xung quanh.
Cy chuyn l qu trnh a vt liu nghin cu hoc canh trng vi sinh
vt vo mi trng thc n mi. Cy chuyn thng nhm mt trong cc mc ch
sau:
- Pht hin s c mt ca vi sinh vt trong vt liu nghin cu
- tng lng vi sinh vt cn thit mt cch nhanh chng.
- Cy chuyn t canh trng c sang canh trng mi gi ging.
2.2.4.2 Phng php m s khun lc mc trn mi trng c [9]
a. Nguyn tc:
Cy mt th tch xc nh mu huyn ph vi sinh vt ln mi trng thch
trong hp petri sau sau mt khong thi gian nui cy tin hnh m khun lc.
b. Tin hnh:
- Pha long ch phm nghin cu
- Cy chuyn ln mi trng trong hp petri
- Da vo s khun lc, th tch mu cy v h s pha long c th
suy ra s khun lc c trong mt ml mu kho st ban u.
Ly 1g ch phm rn v 1ml ch phm lng tin hnh pha long cy trn mi
trng thch trn a petri, bao gi hp petri v lt p li ri nui cy 30
0
C.
i vi nm si sau 24 gi tin hnh m v tin hnh m li trn 48 gi v
nu thi gian nui cy qu di cc khun ty th s pht trin trn khp b mt mi
trng thch .

- 50 -
2.2.4.3 Sn xut ch phm sinh hc
a. Nm mc
Nguyn liu cn sn xut ch phm l pectin, cm go, v tru, amoni
sunfat. m ti u, tit trng 121
0
C trong 15 pht sau ly ra ngui
nhit phng. Nm c cy vo mi trng v sau nui cy nhit
phng.
Bng 2.1: Qu trnh sn xut ch phm Biopep-21, Biopep-22
Tn ch
phm
Thnh phn ha hc (%)
m
(%W)
Thi gian
(gi)
Nhit
(
0
C)
Pectin 9
Tru 15
Cm go 75
Biopep-21
(Aspergillus
niger )
Amoni sunfat 1
56-64 40-44 RT
Pectin 9
Tru 15
Cm go 75
Biopep-22
(Aspergillus
oryzae)
Amoni sunfat 1
56-64 40-44 RT
b. Vi khun
Nguyn liu cn sn xut ch phm l l dch malt, CMC, pectin, amoni
sunfat. iu chnh Brix, pH. Tit trng 121
0
C trong 15 pht sau ly ra
ngui. Vi khun c cy vo mi trng v sau nui cy nhit phng.
Bng 2.2 Qu trnh sn xut ch phm Biopep-31, Biopep-32.
Vi khun
CMC
(%)
Pectin
(%)
Brix pH
Amoni
sunfat
(%)
Thi
gian
(gi)
Nhit

(
0
C)

m
(%W)
Biopep-31
Bacillus polyfermenticus
Biopep-32
Bacillus amyloliquefaciens
0,7 0,7 7-8 5,5 0,09 36 RT 100
- 51 -
2.2.5 Phng php b tr th nghim
Biopep-21,
Biopep-22,
Biopep-31
Biopep-32
Kho st t l bc v ca tiu sau
ln men theo t l ch phm
To ch phm vi sinh vt
Kho st m ca tiu theo
thi gian ngm

Kho st t l bc v ca tiu
sau ln men theo m ht
tiu
Kho st t l bc v ca tiu sau
ln men theo thi gian ln men
Th nghim ln men lp li
i vi Biopep-21, Biopep-22
Kim tra c t aflatoxin,
ochratoxin
Kim tra hm lng vi sinh
TN 1
TN 2
TN 3
TN 4
TN 5
TN 6
TN 7
TN 9
TN 8
W
lm
,TLBV
T l ch phm,
TLBV

Thi gian,
TLBV

Nhit
Cht rn ha
tan, pH
Tiu trng
Aflatoxin,
ochratoxin

W
p
Kho st nhit ln men
Kim tra cht rn ha tan v
pH ca tiu thnh phm
TN 10


Ch thch: TLBV T l bc v
Hnh 2.1 Quy trnh b tr th nghim
- 52 -
2.2.5.1 Kho st m cho qu trnh ln men
Trc khi ln men ngm ht tiu vi thi gian khc nhau v kim tra m.
Cc ht tiu c chuyn sang r nha ro nc. Chuyn tt c tiu vi nhng
m khc nhau vo cc khn ln men v trn u vi mi mu 28% ch phm
sinh hc v ln men trong 48 gi. Sau 48 gi, ly cc mu tiu ln men ra sch
(xem 2.2.1.2) kim tra kh nng bc v v tin hnh phn cp (xem 2.2.1.3). Tnh
s lng ht tiu mi cp v tin hnh so snh gia cc mu c m khc nhau.
(lm 3 ln ly trung bnh). Tin hnh song song lm mu i chng.
2.2.5.2 Kho st t l ch phm sinh hc cho qu trnh ln men
Ht tiu c ngm t c m tt nht cho qu trnh ln men.
Sau chng c chuyn n cc r nha ro nc khong 5 pht. Tin hnh
ln men vi cc t l ch phm khc nhau, trn u ch phm vi ht tiu. Tin hnh
ln men trong 48 gi. Sau 48 gi ln men ly mu ra loi b v ht tiu (xem
2.2.1.2) v phn cp (xem 2.2.1.3). Tnh s lng ht tiu mi cp v tin hnh so
snh gia cc mu c t l ch phm khc nhau.
2.2.5.3 Kho st thi gian cho qu trnh ln men
Ht tiu c ngm t c m tt nht cho qu trnh ln men.
Sau chng c chuyn n cc r nha ro nc. Tin hnh ln men vi t l
ch phm ln men bc v ht tiu tt nht, trn u ch phm vi ht tiu. Tin
hnh ln men nhiu thi gian khc nhau. Sau cc khong thi gian ln men c
nh, ly mu ra loi b v ht tiu v tin hnh phn cp ( xem 2.2.1.3). Tnh s
lng ht tiu mi cp v tin hnh so snh gia cc mu c thi gian ln men
khc nhau. Tin hnh song song lm mu i chng.
2.2.6 Phng php x l s liu:
X l bng excel
-Tm lch chun.
X l bng SG-plus for win
- X l Anova:
- 53 -
Nhp s liu chn compare Analysis of variance One Way Anova
OK
Bng Anova Analysis s xut hin: So snh P-Value vi 0,05. Nu < 0,05 s
tc ng c ngha, >0,05 s tc ng khng c ngha.
- X l multiples Ranges Tests:
Sau khi c kt qu Anova, chn multiples Ranges Tests xut hin bng
cho bit cc gi tr no khc nhau c ngha v gi tr no khc nhau khng c ngha
th hin: cc ch X ng hng khng khc nhau c ngha. Cc ch X khc
hng khc nhau c ngha.











- 54 -
CHNG 3: KT QU V THO LUN
3.1 Kho st m ht tiu theo thi gian ngm.
Kho st m ht tiu bng cch gi nguyn lng nc v trng lng ht
tiu. Thay i thi gian ngm n khi ht tiu t bo ha.
Mi loi tiu c kh nng ht nc khc nhau v vy cn kho st m
bit c thi gian ngm tiu t c m cn thit cho qu trnh ln men.
M TNG THCH VI THI GIAN NGM
0
10
20
30
40
50
60
70
0 100 200 300 400 500 600 700 800
THI GIAN (PHT)

M

(
%
)

th 3.1: m ht tiu tng thch vi thi gian ngm.
Tiu en ht nc rt chm, sau 765 pht (12 gi 75 pht) tiu t m
bo ha 64,40 %. Ban u tiu ht nc rt nhanh sau 45 pht tiu t m ban
u l 10,7% tiu t m 38,50% v tng m u dn dn sau 225 pht. Sau
m tng chm dn t n bo ha (12 gi).
ht nc ca tiu l kh nng ht v gi nc ca ht tiu, iu kin
thng v c xc nh bng cch ngm mu vo trong nc c nhit t 20-
30
0
C. Trong iu kin nc ch c th chui vo nhng l rng h ca ht tiu.
bo ho nc ca ht tiu l ht nc cc i ca tiu khi p sut trong ht
tiu bng p lc xung quanh, ht tiu khng th hp thu nc c na, im ny
gi l bo ha m ca ht tiu.
- 55 -
3.2 m ht tiu trong qu trnh ln men cho t l bc v cao nht
Sau khi ln men tin hnh phn cp v kim tra, tnh c t l bc v ht
tiu (xem ph lc D.1). Ch phm sinh hc ln men b nh hng bi nhiu yu t,
trong yu t m l quan trng nh hng n s pht trin ca nm, mi loi
nm thch hp vi mt ngng m khc nhau. m qu thp hay qu cao u
c ch s sinh trng v pht trin ca vi sinh vt. Ngoi ra cc phn ng thy phn
gia pectin v celullose vi petinase v cellulase cn s hin din ca nc. C
nh thi gian v t l ch phm, thay i m, kim tra kh nng ln men bc v
ht tiu bit c m thch hp cho s pht trin ca vi sinh vt.
th 3.2 T l bc v tng thch m ln men (Biopep-21, Biopep-22)
Qua biu 3.2 thy t l bc v thay i khc nhau ty theo m. V mi
loi nm pht trin mt ngng nhit khc nhau. m m nm pht trin tt
nht gi l m ti u. Khi ch phm c trn vi cc mu tiu c m khc
nhau, da vo kh nng ln men bc v ht tiu ta tm c m tt nht cho s
pht trin ca nm. Khi nm pht trin pht trin trn b mt ht tiu chng tit ra
T L BC V TNG THCH M LN MEN
20
30
40
50
60
70
80
90
30 35 40 45 50 55 60
M (%)
T

C

V

(
%
)
Biopep 22 Biopep 21 i chng
- 56 -
enzym pectinase v cellulase c kh nng phn gii pectin, cellulose thnh cc dng
carbohydrate n gin to cht dinh dng hay ngun thc n cho s pht trin ca
nm mc. Khi pectin v cellulose b phn gii s lm v ht tiu c bc ra d
dng hn.
ng vi m 35% Biopep-21 bc v c 53,30%, tip tc tng m
43%, 45%, 46%, 49% n 55% t l bc v tng ng l 61,17%, 61,91% v t
phn trm t l bc v cao nht m 46% l 76,96% .
So snh Biopep-22 vi m 35% t l bc v 73,37%, tng m dn ln
43%, 45%, 46%, 49% n 55% t l bc v tng ng l 74,46%, 75,51%, 78,10%.
V m 49 % mu tiu t t l bc v cao nht 82,61%.
Cc mu ln men th nghim c chia thnh cc mu c m 35%, 43%,
45%, 46%, 49%, 51%, 53%, 54%, 55% tm m ti u cho nm pht trin.
i vi cc mu i chng khng c ch phm ht tiu vn c bc v nhng vic
bc v rt kh khn. Ht tiu c bc v nh vo hp thu nc lm cho v ht
tiu mm v da vo lc ch xt bng tay. Mc d vn c c tiu s nhng mu
sc ht tiu khng trng v cn rt nhiu ht bc v mc 25%, 50% hay 75%.
Mu i chng c t l bc v cao khi tng m n 55% t l bc v 44,26%.
(Xem ph lc D, bng D.1. D.2, D.3)
Ht tiu c ln men bng Biopep-21 t c t l bc v cao nht ti
m 46% vi t l tiu trng l 76,96% so vi mu i chng khng c ch phm
vic s dng Biopep-21 sn xut ra enzym pectinase v cellulase phn gii pectin
v cellulose lm v ht tiu c bc ra d dng v gn nh hon ton. V vy
m ca ht tiu ln men bng Biopep-21 c chn cho th nghim sau l 46%.
Ln men bng ch phm Biopep-22 t l bc v cao hn v tt hn c v
phn trm bc v v mu sc tiu trng. Ti m 49 % t l bc v t cao nht
82,61%. Ti m ny Biopep-22 ln men rt tt, iu ny chng t trong qu trnh
pht trin chng tit ra nhiu pectinase v cellulase phn gii pectin v cellulose
nng cao t l sn xut tiu trng. m 49% c chn l m tt nht cho qu
trnh ln men ca Biopep-22.
- 57 -
3.3 T l ch phm cho qu trnh ln men ht tiu t t l bc v cao nht.
3.3.1 Ch phm Biopep-21, Biopep-22
Mc ch: Chn c t l ch phm thch hp nht cho qu trnh ln men (xem
ph lc E, F).
Yu t c nh: Biopep-21 tiu c m 46%
Biopep-22 tiu c m 49%
Thi gian ln men : 48 gi
Yu t kho st: T l ch phm 20%, 24%, 28%, 32%, 36%, 40%
T L BC V TNG THCH T L CH PHM
40
50
60
70
80
90
100
16 20 24 28 32 36 40 44
T L CH PHM (%)
T

C

V

(
%
)
Biopep-21 Biopep-22

th 3.3: T l bc v tng thch vi t l ch phm Biopep-21, Biopep-22
T l bc v tng dn khi tng t l ch phm, ch phm Biopep-22, Biopep-
21 ban u t l ch phm 20% t l bc v tng ng l 73,17% v 53,30%.
Biopep-22 t cao nht khi t l ch phm 32% t l bc v l 87,64 %. Biopep-21 l
34% v t l bc v cao nht 83,91%. Khi tip tc tng t l ch phm th t l bc
v ht tiu li gim khng th tng hn na. m tt nht cho qu trnh ln men
c chn, v vy cn chn t l ch phm ph hp bc v ht tiu c ti
a. (Xem ph lc D, Bng D.4, D.5)
- 58 -
Khi b sung ch phm t, nm tng trng v pht trin tt nhng lng
enzym pectinase v cellulase khng bc v ht tiu hon ton v tt nht. Khi
t l ch phm c b sung qu nhiu, nm pht trin nhiu trn b mt ht tiu ly
cht dinh dng cho s sinh trng v pht trin ca nm, s khng cung cp
cho nm pht trin. Mt khc, m thch hp ln men b gim nhiu do nm s
dng. Nh vy nm khng m tt nht cho s pht trin dn n nm b c
ch s sinh trng. Kh nng tit enzym pectinase v cellulase thp khng phn
gii pectin v cellulosec ca v ht tiu.
V vy vi cc th nghim v kt qu trn, m 49% t l ch phm l 32%
Biopep-22 cho kh nng ln men tt nht vi t l bc v 87,64%. Biopep-21 vi
m 46% , t l ch phm 36% t l bc v cao nht l 83,94% c chn cho th
nghim sau.
3.3.2 Ch phm Biopep-31 v Biopep-32
Tin hnh th nghim
Mc ch: Kho st tc ng ca t l ch phm Biopep-31 v Biopep-32
n t l bc v ht tiu.
Yu t c nh:
Khi lng tiu: 50 g
Thi gian ln men: 72 gi
Yu t kho st :
T l ch phm: 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%
- 59 -
Bng 3.1: Kho st t l ch phm Biopep-31, Biopep-32 n t l bc v ht
tiu.











T L BC V TNG THCH T L CH PHM (Biopep-31, Biopep-32)
45
48
51
54
57
60
63
0 10 20 30 40 50 60
T L CH PHM (%)
T

C

V

(
%
)
Biopep-31 Biopep-32

th 3.4: T l bc v tng thch vi t l ch phm Biopep-31, Biopep-32
Da vo kt qu phn tch anova
S tc ng ca t l ch phm ln t v bc v ht tiu c tc ng v s tc
ng ny khc bit c ngha thng k mc tin cy 95% (xem ph lc E, F)
Biopep-31

Biopep-32

T l ch phm (%)
T l bc v T l bc v
10 48.341,69 47.460,69
15 49.580,94 49.620,48
20 50.710,58 50.720,57
25 51.521,26 53.690,86
30 56.290,55 56.020,06
35 54.321,10 57.010,41
40 52.641,65 60.260,16
45 50.251,52 56.710,74
50.00 49.521,96 53.280,18
- 60 -
V y l ch phm lng nn khng cn kho st yu t m. Xc nh t l
ch phm ln men thy rng t l bc v cao hn so vi mu i chng khng
nhiu nhng tiu bc v tt hn, mu sc trng sng.
Kt qu: Biopep-31 t l ch phm 30% t l bc v 56,29%
Biopep-32 t l ch phm 40% t l bc v 60,26%
3.4 Thi gian ln men ht tiu t t l bc v cao nht
3.4.1 Ch phm Biopep-21, Biopep-22
C nh m v t l ch phm c chn (xem 3.2 v 3.3), thay i
thi gian ln men. Tm thi gian ln men tt nht cho s pht trin ca nm mc v
t l bc v ht tiu l cao nht.
Cn kho st thi gian ln men v nu thi gian ln men qu ngn hot tnh
enzym pectinase v cellulase cha tip xc ti a vi c cht tng thch, cn nu
thi gian ln men qu di sau khi phn gii ht pectin v cellulose cu thnh nn
vch t bo ht tiu chng s tip tc s dng cc cht ha tan lm cht dinh dng
pht trin lm gim cht lng ht tiu. Mt khc mi vi sinh vt c mt thi
gian pht trin mnh nht ti hot tnh enzym pectinase v cellulase l cao nht.
hn na khi nm pht trin mt thi gian di trn ht tiu c th sinh ra c t gy
hi cho sc khe ngi s dng.
T L BC V TNG THCH THI GIAN LN MEN
20
30
40
50
60
70
80
90
100
110
24 36 48 60 72 84 96 108 120 132
THI GIAN LN MEN (h)
T

C

V

(
%
)
Biopep-22 Biopep-21
i chng ( Biopep-21) i chng (Biopep-22)

th 3.5: T l bc v tng thch thi gian ln men
- 61 -
T l bc v tng dn khi tng thi gian ln men t 24 gi lc ny v ht tiu
bt u bc ra d dng hn nhng nht cn cao do vn cn pectin, v cha
bc v c hon ton, tip tc tng thi gian n 72 gi (3 ngy), ch phm
Biopep-22, Biopep-21 t l bc v tng ng l 99,72% v 99,56% lc ny tiu rt
d loi b v v ht tiu trng. Cc phn trm cn li l ht tiu cn non cha s,
tiu xu, nhn, v tiu bc v nhng mu sc v cht lng tiu khng tt,khng
s dng c.
Kt qu sau kho st s dng ch phm Biopep-21, Biopep-22 ln men sn
xut tiu trng sau 3 ngy ht tiu c bc v hon ton, mu sc trng ng. So
vi cc thi gian khc ht tiu cng c bc v hon ton nhng m bo cht
lng ht tiu cch kim tra cht rn ha tan sau 3 ngy l cao nht v tit kim thi
gian sn xut cho tnh kinh t v vy chn thi gian ln men cho ch phm Biopep-
21, Biopep-22 l 3 ngy.
3.4.2 Ch phm Biopep-31 v Biopep-32
Mc ch: Tm thi gian m hot tnh pectinase v cellulase tc ng ln c
cht mnh nht v t l bc v l cao nht.
Yu t c nh: T l ch phm Biopep-31 l 30%
T l ch phm Biopep-32 l 40%
Yu t kho st: Thi gian ln men 3 ngy, 4 ngy, 5 ngy, 6 ngy, 7 ngy
Bng 3.2: T l bc v tng thch thi gian ln men ca Biopep-31, Biopep-32







Biopep-31 Biopep-32
i chng
Thi gian
(ngy) T l bc v (%)
3 56.550.61 60.400.15 43.120.26
4 58.320,76 61.592,47 49.200.69
5 64.440,29 63.831,64 55.191.20
6 64.580,91 68.840,94 59.391.40
7 64.480,34 68.920,13 59.911.22
- 62 -
T L BC V TNG THCH THI GIAN LN MEN (Bi opep-31, Bi opep-32)
35
40
45
50
55
60
65
70
75
2 3 4 5 6 7 8
THI GIAN (NGY)
T

C

V

(
%
)
Biopep-31 Biopep-32 i chng

th 3.6: T l bc v tng thch thi gian ln men Biopep-31, Biopep-32
Mi vi sinh vt c mt thi k pht trin mnh nht ti hat tnh enzym
pectinase v cellulase tc ng mnh m ln c cht, cn kho st ti thi gian no
t l bc v l cao nht. Biopep-31 t l bc v cao nht l 5 ngy 64,58%, Biopep-
32 l 6 ngy t l bc v 68,84%.
So vi mu i chng khng s dng ch phm, mu c s dng ch phm t
l bc v cao hn, mu sc ht tiu trng hn v v c bc hon ton mt cch d
dng.
Khi s dng ch phm Biopep-31, Biopep-32 ln men sn xut tiu trng t
l bc v vn cn thp v thi gian sn xut kh di nu p dng thc t khng c
tnh kinh t v vy chng ti ngng kho st ti y.
3.5. Kim tra gi tr pH v cht rn ha tan ca ht tiu ln men
Gi tr pH ca ht tiu ln men thay i trong khong 6,0-6,55 (bng 3.3).
Tiu ln men bng ch phm Biopep-21, Biopep-22 gi tr pH tng nh v sau
gim dn. Sau 3 ngy tiu trng Biopep-21 c pH= 6,51, Biopep-22 c gi tr pH=
6,39. Tuy nhin gi tr pH t 6 n gn trung tnh khng nh hng n sc khe
ca con ngi v hng v cc mn n.


- 63 -

Bng 3.3 : Kim tra gi tr pH v cht rn ha tan ca ht tiu ln men
Tn ch phm Thi gian ln men (h)
0 24 36 48 60 72 84 96 108 120
Biopep-21 6,21 6,30 6,32 6,35 6,45 6,51 6,55 6,48 6,34 6,00
Biopep-22
pH
6,21 6,46 6,43 6,40 6,39 6,39 6,36 6,25 6,19 6,16

GI TR pH TNG THCH THI GIAN LN MEN
5.9
6
6.1
6.2
6.3
6.4
6.5
6.6
0 12 24 36 48 60 72 84 96 108 120 132
THI GIAN (gi)
p
H
Biopep-21 Biopep-22

th 3.7: pH tng thch thi gian ln men
Cc thc phm khi ln men thng b chua v gi tr pH qu thp s nh
hng n sc khe con ngi. V vy sau khi kim tra pH tiu trng sau ln men,
sn phm khng mang tnh axit v khng nh hng n sc khe con ngi, c th
s dng trong cc mn n m khng lm gim cht lng v hng v mn n.
Bng 3.4: Kt qu kim tra hm lng cht rn ha tan ht tiu ln men


Cht rn ha tan theo thi gian ln men
Tn ch phm
Thi
gian
(gi)

24 36 48 60 72 84 96 108 120
Biopep-21 0,18 0,18 0,18 0,18 0,19 0,18 0,17 0,17 0,17
Biopep-22
Bx
0,19 0,2 0,19 0,2 0,21 0,2 0,19 0,18 0,18
- 64 -
Sau khi lp pectin v cellulose b phn gii hm lng cht rn ha tan d
thot ra ngoi lm tng hm lng cht rn ha tan nng cao hng v khi s dng.
V vy cn ti u cc iu kin ln men v nu lng ch phm sinh hc qu nhiu
hay thi gian ln men di, vi sinh vt sau khi phn gii c cht pectin v cellulose
s dng ht s tip tc vi ngun dinh dng mi l cht rn ha tan. Kt qu
sau 3 ngy hm lng cht rn ha tan t cao nht.
3.6 Kho st s thay i thay i nhit theo thi gian ln men
Nhit thch hp cho s pht trin ca nm mc dao ng xung quanh
30
0
C. Sau mt ngy ln men nhit khi bt u tng dn sau 84 gi nhit
ln men gim dn xung.
Bng 3.5: Nhit thay i theo thi gian ln men.
Tn ch
phm
Thi gian ln men (h)

0 24 36 48 60 72 84 96 108 120
Biopep-21

T
0
C
28 30 31 31,5 31 30 29,5 29,5 29 29
Biopep-22

28 30,5 32,5 32 31,5 31.5 31 30,5 29,5 29

NHIT TNG THCH THI GIAN LN MEN
27
28
29
30
31
32
33
0 12 24 36 48 60 72 84 96 108 120 132
THI GIAN (gi)
N
H
I

(
C
)
Biopep-21 Biopep-22

th 3.8: Nhit tng thch thi gian ln men
- 65 -
Nhit qu trnh ln men tng cao l do s tng trng mnh m ca nm,
trong qu trnh ln men chng sinh ra nng lng lm tng nhit khi . Khi n
mt giai on nm ngng pht trin nhit khi s gim. Nhit thch hp cho
nm pht trin t 22-35
0
C. Nu nhit cao hn hoc thp hn nh hng khng
tt n s pht trin ca nm mc. Nhit ln men c kim tra thch hp cho
s pht trin ca nm mc. Sau 48 gi nhit ln men Biopep-21 cao nht 31,5
0
C,
Biopep-22 sau 36 gi l 32,5
0
C. Sau 3 ngy nhit vn thch hp cho s pht trin
ln men ca nm mc.
3.7 Kt qu kho st s lng t bo vi sinh vt ban u v xc nh hot tnh
enzym ca cc ch phm.
Bng 3.6: Kt qu kho st s lng t bo vi sinh vt ban u v xc nh
hot tnh enzym ca cc ch phm.

3.8 Kt qu kim tra piperine
Tiu en nguyn liu sau khi c ln men c kim tra nh lng
piperine bi Cng Ty C Phn Dch V Khoa Hc Cng Ngh Sc K Hi ng
(79 Trng nh, Qun 1, Thnh ph H Ch Minh, Vit Nam)
Ch phm
Hot tnh
cellulase
(IU/g)
Hot tnh
pectinase
(IU/g)
S t bo vi
sinh vt
(CFU/ml)
Biopep-21(sn xut t
Aspergillus niger)
59,8 125,68
1,43.10
11
Biopep-22
( Aspergyllus oryzae )
51,2 198,75
1,625.10
11
Biopep-31
(Bacillus polyfermenticus)
59,8 118,143
725.10
6
Biopep-32
(Bacillus amyloliquefaciens)
53,2 121,72
715.10
6
- 66 -
Bng 3.7 nh lng piperine trong tiu trng
Mu Phng php th nh lng piperine (%)
Tiu en nguyn liu 3.7
Tiu ln men bi Biopep-21 5.6
Tiu ln men bi Biopep-22
ISO 5564
5,7

Piperine l thnh phn ha hc rt quan trng to v cay cho ht tiu.
Piperine c tc dng hu cho sc khe bao gm gim cn, tng cng hp thu cht
dinh dng, chng nhim trng v chng ung th. Piperine c chit xut t ht
tiu en nguyn liu theo kt qu phn tch 3,7%, cht rn ha tan (bao gm c
piperine) khng th c chit xut hon ton t cc t bo ht tiu v cu to nn
thnh t bo rt vng chc lm ngn cn vic chit xut cc cht, trong cellulose
v pectin l hai thnh phn chnh. S dng ch phm sinh hc (cha cc vi sinh vt)
c th tng hp enzym cellulase v pectinase phn gii cc cht nh cellulose v
pectin thnh t bo ht tiu gii phng cc cht rn ha tan hon ton.
Kt qu cho iu ny hm lng piperine tng 51,35% khi s dng Biopep-21,
Biopep-22 tng 54,05%. Piperine d c khuch tn khi s dng iu ny chng
minh ti sao khi s dng tiu s lun cay hn so vi tiu en.
3.9 Kim tra c t sn phm tiu sau ln men (aflatoxin, ochratoxin A)
Tiu en nguyn liu sau khi c ln men c kim tra c t aflatoxin ti
Vin Pasteur Thnh Ph H ch minh (167 Pasteur, Qun 3, Thnh Ph H Ch
Minh)
Bng 3.8. Kt qu kim tra aflatoxin ca tiu trng
Mu Phng php n v Aflatoxin
Tiu ln men bi Biopep-21
Tiu ln men bi Biopep-22
HPLC ppb Khng pht hin
- 67 -
Tiu en nguyn liu sau khi c ln men c kim tra c t Ochratoxin
A bi Cng Ty C Phn Dch V Khoa Hc Cng Ngh Sc K Hi ng (79
Trng nh, Qun 1, Thnh ph H Ch Minh, Vit Nam)
Bng 3.9 Kt qu kim tra ochratoxin A ca tiu trng
Mu Phng php n v Ochratoxin A
Tiu ln men bi Biopep-21
Tiu ln men bi Biopep-22
TK.AOAC 2004.10 %
Khng pht hin
(LOD=1)

Hai ch phm Biopep-21, Biopep-22 u khng sinh ra c t aflatoxin
ochratoxin A trong sut thi gian ln men. V vy chng ta c th p dng vo sn
xut v s dng tiu trng vo ch bin thc phm.
3.10 Kim tra yu t vi sinh vt c hi trong sn phm tiu sau ln men
m bo cht lng an ton v sinh thc phm ch tiu vi sinh vt c
kim tra m bo rng trong qu trnh thc hin sn phm vn an ton cho ngi s
dng.
Bng 3.10 Kt qu kim tra vi sinh ca tiu trng
Tiu trng ln men Th nghim PP th nghim Gii hn Kt qu
Biopep-21 v Clostridium perfringer ISO 7937:2004
Biopep-22 Staphylococci coaglulase ISO 6888-1:1999
10 Cfu/g <10


- 68 -
3.11 S quy trnh to ch phm Biopep-21, Biopep-22
































Hnh 3.1: S quy trnh to ch phm Biopep-21, Biopep-22








Thi gian: 40-44

gi
m: 56-64%
Nhit : RT
Mi trng
thch nghing
Cy chuyn
Nui RT
Trn u bo t
Cy chuyn
Nui
ng mc gc
Nc v trng
Xay
Ch phm
Bt go
Mi trng rn
T l: 1:1
- 69 -
3.12. Quy trnh sn xut ch phm Biopep-31, Biopep-32



Hnh 3.2 Quy trnh sn xut ch phm Biopep-31, Biopep-32


Bng 3.11: Kt qu kho st t c sau qu trnh ln men (Biopep-21, Biopep-22)
Ch phm
m
(%)
T l ch phm
(%)
Thi gian ln men
(ngy)
T l bc v
(%)
Biopep-21 46 36 99,56
Biopep-22 49 32
3
99,72
Mi trng
thch nghing
Cy chuyn
Nui RT
Trn u bo t
Cy chuyn
Nui
ng mc gc
Nc v trng
Ch phm
Mi trng lng
Thi gian: 36 gi
Nhit : RT
- 70 -
3.13. Quy trnh sn xut tiu trng s dng ch phm sinh hc Biopep-21,
Biopep-22



































Hnh 3.3 Quy trnh sn xut tiu trng s dng ch phm sinh hc Biopep-21,
Biopep-22
Ch thch: W: m, TLCP: t l ch phm
Ch
Sng
ng gi
La chn
Ra
Ln men
Ngm
Sy
Tiu
trng
Biopep-21: W 46%
Biopep-22: W 49%
Thi gian: 3 ngy
Biopep-21: 36% TLCP
Biopep-22: 32% TLCP
Nhit :55-60
0
C
W: 12%-13%
Tiu nguyn
liu
- 71 -
Sn phm t t l bc v trn 99%, khng c c t, hm lng piperine cao
v an ton cho ngi s dng.
Bng 3.12: Kt qu t c c th p dng vo quy m sn xut cng nghip
Sn phm pH
Piperine
(%)
Aflatoxin,
Ochratoxin A
Ch tiu vi
sinh
Tiu ln men bi Biopep-21
6,51 5,6
Tiu ln men bi Biopep-22
6,39 5,7
Khng c t

Kt qu t c sn phm tiu trng sn xut bng ch phm sinh hc.
Mi v: Khi nghin thnh bt c mi thm c trng ca ht tiu trng, cay v
khng c mi v v l.
Mu sc: Mu ca ht tiu trng t xm nu nh n hi trng ng.
Hnh dng: Vn gi nguyn hnh dng ca ht tiu.
Kch thc : ng u.
Cu trc : Cng, gin
Da vo kt qu t c chng ti tin hnh ln men vi s lng ln hn
1Kg tiu/m. Kt qu vn t c nh trn.
Bng 3.13 So snh kt qu t c vi cc ti khc.
Ni dung
Vaidyanatha Lyer
Thankamani and
Raghavan Nair
Giridhar (2004)
PGS.TS
Nguyn
c
Lng
(2004)
K s Bi
Thanh Ha
(2010)
K s
V Th Ngc
Thanh (2010)
Sinh vin Nguyn Th
M Hin, Nguyen Van
Hoang (2010)
S dng
vi sinh
vt
B. mycoides
B. licheniformis
B. brevis
A. niger
Biocoffee-3
(A. phoenicis)
Biopep-6
(T. piluliferum)
Biopep-21
(A. niger)
Biopep-22
(A.oryzae)
T l ch
phm
_ _ 35% 25% 36% 32%
Thi gian
ln men
4 ngy 4 ngy 3 ngy 3 ngy 3 ngy 3 ngy
nh
lng
piperine
4% _ 5% 5,7% 5,6% 5,7%

- 72 -
KT LUN V KIN NGH
A. Kt lun
Xy dng quy trnh sn xut sn xut ch phm Biopep-21, Biopep-22,
Biopep-31, Biopep-32 ( Xem 3.11-3.12)
Xy dng thnh cng quy trnh sn xut sn xut tiu trng s dng ch
phm Biopep-21, Biopep-22. (Xem 3.13)
Sn phm khng c c t, an ton cho ngi s dng.
Hm lng piperine chit xut tng 51,35% khi s dng Biopep-21, Biopep-
22 tng 54,05%.
Nghin cu thnh cng hai ch phm Biopep-21 (c sn xut t
Aspergillus niger), Biopep-22 (Aspergillus oryza) dng sn xut tiu trng
tng hm lng cht tan (piperine) ng gp vo s pht trin ca ngnh
cng ngh ha thc phm.
Cha thnh cng vi hai ch phm Biopep-31 (Bacillus polyfermenticus),
Biopep-32 (Bacillus amyloliquefaciens.)
B. Kin ngh
c th sn xut tiu trng quy m ln hn cn c quy trnh lm tinh ch
phm trc khi s dng mi bo m c cht lng ch phm v ko di
thi gian bo qun.
Cn kho st nghin cu thm v ch phm Biopep-31
(Bacillus polyfermenticus), Biopep-32 (Bacillus amyloliquefacien).
Tin hnh nghin cu phi hp gia cc ch phm Biopep-21, Biopep-22,
Biopep-31, Biopep-31 c ngun gc t Aspergillus niger, Aspergillus
oryzae, Bacillus polyfermenticus, Bacillus amyloliquefaciens tng ng
c phn lp t vn tiu.