Tổng Quan Về Collagen Và Ứng Dụng Của Collagen

AMH CNG NGH THC PHAM GVHD: TS.
PHAN NGC HA
SVTH: V TH HONG LI NH Trang 1

CHNG 1; COLLAGEN
1.1 GII THIU CHUNG V COLLAGEN
1.1.1 Khi qut v Collagen [2]
Collagen l mt protein c cu trc bc 4 in hnh, do cc n v tropocollagen cu trc
bc 3 t hp theo cc hng dc v ngang lm collagen c nhiu mc cu trc. Collagen l loi protein
cu trc chnh yu, chim khong 25% - 35% tng lng protein trong c th cc ng vt c xng
sng. Collagen c phn b" trong cc b phn nh da, c, gn, sn, rng, h thng mch mu ca ng
vt v c mt trong cc lp mng lin kt bao quanh cc c v l thnh phn chnh ca dy chng v
gn,... Khong 10% protein trong c ng vt c v l collagen, cc protein ngoi bo (hn 90% trong
gn, xng v khong 50% trong da) c cha collagen.
Trong thnh phn ca da, collagen chim khong 70% cu trc da v c phn b" ch
yu lp h b ca da, to ra mt h thng nng , h tr cc c tnh c hc ca da nh : sc cng,
n hi, duy tr m,... N ng vai tr kt ni t bo, kch thch qu trnh trao i cht, to n hi
cho da. S suy gim v s" lng v cht lng collagen s dn n hu qu lo ha ca c th m s
thay i trn ln da, khun mt l du hiu d nhn bit nht: ln da b kh, nhn nheo bt u t cc
ng nhn mnh trn khe mt, khe ming, lu dn thnh np nhn su, cc ng nt khun mt b
chng nho v chy x. Ty theo tng tui, iu kin sng v tc ng ca mi trng, da c th b lo
ha hoc tn thng , khi si collagen s mt dn tnh n hi v sn chc do cu trc collagen b ph
hy. Chnh v vy m collagen ng vai tr l mt trong nhng cht quan trng hng u ca ngnh thm
m, c bit l chm sc da, phu thut thm m, phu thut bng...
Collagen cng c phn b" gic mc, v tn ti di dng tinh th. Trong m c,
collagen chim khong t 1% n 2%. Trong c th ngi, collagen chim t 20 - 25% protein ca c
th.
Collagen l thnh phn chnh ca m lin kt, c chc nng to vng chc
v n hi. Collagen c tc dng ging nh mt cht keo lin kt cc t bo li vi nhau hnh thnh cc
m v c quan nn tng trong c th. Collagen cung cp cho cc m lin kt nhng c tnh ni tri nh
vo s hin din rng khp v s sp xp mang tnh cu trc ca n. Theo quan im y sinh, collagen
AMH CNG NGH THC PHAM GVHD: TS. PHAN NGC HA

ng vai tr quan trng trong s pht trin, hn gn vt thng, l thnh phn hot ho cc tiu cu trong
mu v s hnh thnh mch. Thm vo s pht trin ca mt s" bnh di truyn c xc nh l do s
t bin trong cu trc gen ca collagen. Nhng r lon trong qu trnh tng hp hoc phn gii collagen
u c ghi li nh l nguyn nhn ca nhiu cn bnh phc tp nh chng vim khp, x gan, bnh i
ng v ung th cng nh nhng bnh c lin quan n s lo ho. Collagen phn b" khp ni trong c
th, t ch gn ni bp chn vi gt chn cho ti gic mc. Trong gn v dy chng, collagen c tc dng
truyn lc t c sang xng v tch tr nng lng n hi. Chnh nh tnh cht ny m c th c th di
chuyn, vn ng mt cch nhp nhng v uyn chuyn. Collagen cn l cht nn hu c c trong xng
v men rng gip chng chng li s rn nt. N l thnh phn chnh trong da, mch mu, cc c. Bn
cnh cc chc nng c hc, gic mc, thy tinh th, trt t cu trc ca cc si collagen to nn s trong
sut duy tr th lc.
Collagen c xem nh mt vt liu mang tnh xy dng. S linh hot ca n l nh
vo cu trc cp bc phc tp, to nn s a dng trong tnh cht nhm phc v nhng chc nng nht
nh. Xt v mt kinh t, ngun nguyn liu giu collagen dng sn xut ra gelatine- cht c nhiu ng
dng trong nhiu lnh vc khc nhau: thc phm, y hc v dc phm, m phm, cng ngh nhip nh,...
1.1.2 Lich s pht trin ca Collagen
Theo ting Hy Lp, collagen c ngha l ngi sn xut keo h, ni n qu trnh nu da v
gn ca nga cng nhng loi ng vt khc thu c h. Keo dn collagen c ngi Ai Cp s
dng cch y 4000 nm v ngi M s dng n cch y khong 1500 nm.
T hn 2000 nm trc, con ngi bit s dng m lin kt v sn phm ca n trong ch
bin thc phm to ra cc sn phm dng gel, cc cht dnh. Ngi Ai Cp c i sn xut ra c
collagen v nhng nghin cu cho thy trong nhng th k tip theo tht s xut hin collagen trong
cc ba tic, v d trong mn c hi hay bnh hoa qu,...
Nm 1682, mt ngi Php tn Papin cng b" mt kt qu l thu c mt hn hp
ging jelly t xng. T nm 1700, thut ng collagen tr nn thng dng.
Nm 1754, bi bo u tin trong lnh vc cht dnh c cng nhn Anh v vic sn xut

cht dn ca mt ngi th lm g. Cc cht h dn t nhin c sn xut trn thnh phn c bn l
collagen v mt vi cht khc.
Nm 1850, cng nghip sn xut collagen xut hin ti M, ngun nguyn liu chnh lc ny
l xng.
Nm 1871, mt pht hin quan trng ca bc s ngi Anh Richard Leach Maddox v ng
dng ca collagen a n bc t ph trong ngnh cng nghip hnh nh, v bc s ci tin dry
plate vi mt lp gelatin - bc bromua v cn nhy hn khi so vi wet plate . Sau nghin cu ca
Charles Benne tip tc hon thin phng php dng dry plate. u im ln nht ca phng php
ny l gim rt nhiu thi gian trong cng nghip nhip nh.
Cui th k 19, khi ngnh cng nghip sn xut collagen xut hin lm tng thm cc ng
dng ca collagen trong cc ngnh cng nghip khc v n nh tnh cht sn phm.
Nm 1930, ngnh sn xut collagen c pht trin mnh m khi da heo c xem nh l
mt ngun nguyn liu thu nhn collagen. Ngnh cng nghip sn xut collagen Chu u ch mi bt
u khong nhng nm 1930 nhng Chu u li tr thnh khu vc sn xut collagen quan trng nht trn
th gii.
Nm 1950, cng nghip sn xut collagen c nhng bc pht trin ng k v k thut, t
l nn tng cho cc tiu chun trong sn xut v cht lng sn phm ngy nay.
Ngy nay, s hiu bit v acid amin v protein ni chung, collagen cng nh gelatin ni ring
ngy cng nhiu. Thm vo l nhng tin b trong k thut gip sn phm collagen t an ton v
sinh, cht lng theo tiu chun quc t v yu cu nghim ngt ca cc yu t" k thut.
1.1.3 Ngun gc thu nhn Collagen
Trong nhng nm trc y, p ng nhu cu trong cng nghip, collagen c trch
ly ch yu t da, xng ca cc loi gia sc v ln. Khong thi gian gn y, s bng pht ca cc loi
bnh truyn nhim nh bnh b in (Bovine Spongiform Encephalopathy - BSE, Tranmissible
Spongiform Encephalopathy - TSE) v bnh l mm long mng (Food and Mouth Disease - FMD) ln
v gia sc hn ch phm vi s dng ca collagen c ngun gc t nhng loi ng vt trn bi v

nhng bnh truyn nhim c kh nng ly truyn sang con ngi thng qua cc m ca ng vt. Bn
cnh , mt s" quc gia, collagen trch ly t ln khng c s dng v ro cn tn gio. Vi nhng
nguyn nhn trn, cc nh khoa hc ang tp trung vo cc nghin cu ca h tm ra nhng ngun
collagen thay th. Da, xng, vy, vy ca c nhng loi c nc ngt, c bit l c da trn: c tra, c ba
sa, c bin, da g, da ch, da mc,...c th c s dng nh nhng ngun thay
Trong s" nhng ngun thay th, c cung cp mt ngun nguyn liu th tt
nht v:
- D tm, sn c s dng
- Khng c s ly truyn bnh
- Khng gp phi tr ngi v mt tn gio.
- C kh nng thu c collagen vi hiu sut cao
Khong 70% tng trng lng c th ca c b b i di dng cc ph phm nh da, xng,
vy, u, vy, rut,...trong sut qu trnh ch bin. Vic tn dng nhng cht thi ny c th nng gi tr
kinh t ca cc loi c ln.
Da vo thnh phn % ca tng lng collagen tnh trn tng lng protein, cc loi
c c chia thnh 3 nhm:
- Nhm c hm lng collagen thp: tng lng
collagen chim
n 5 % so vi tng lng protein.
- Nhm c hm lng collagen trung bnh : tng
lng collagen
chim t 5 -- 10 % tng lng protein.
- Nhm c hm lng collagen cao: tng lng
collagen trn
10 % tng lng protein.

Tht ca nhng loi c khc nhau cha hm lng khc nhau ca collagen ha tan trong acid
(acid soluble collagen) v collagen khng ha tan (insoluble collagen). Hm lng collagen tng cng
dao ng t 0,3 -r 2,99 % so vi trng lng ti ca cc m, tng ng vi phm vi t 1,58 4- 13,39 %
so vi tng lng protein.
Bng 1.1 Thnh phn collagen trong tht c

Loi c
Collagen ha tan trong acid
(so vi trng lng ti)
Collagen khng hatan (so
vi trng lngti)
Collagen tng (so vi
trng lngti)
Collagen tng (so vi
protein tng)
C chim
trng
0,19 0,11 0,3 1,58
Sardine 0,36 0,09 0,45 2,42
C thu 0,38 0,09 0,47 2,41
C bn 0,4 0,18 0,58 2,98
C chp 0,49 0,21 0,7 3,25
C i 0,94 0,25 1,19 5,89
C ng 1,06 0,39 1,45 7,35
C mp 2,13 0,86 2,99 13,11
C ui 2,3 0,5 2,8 13,39

Tng lng collagen trong c th thay i t 3,26 % ( c thu) cho n 6,97% ( c
chnh Nht Bn). Lng collagen c th ha tan trong acid (acid soluble collagen) nm trong khong t
13,1 % (c chnh) n 56,6 % (c bn). Hm lng collagen thu c s thay i ty theo loi v loi m
c s dng trong qu trnh trch ly. Trong tt c cc loi, kh nng ha tan ca collagen trong tht
tng i cao nhng thp i vi nhng loi collagen trong ni tng. Collagen trong da, vy, xng v
vy chim phn ln trong tng lng Collagen.
Cho n nay, hu ht cc loi m phm trn nn collagen u c cha protein chit xut t
gia sc. Bi s bc pht ca bnh b in BSE (Bovine Spongiform Encephalopathy - mad cow disease),
collagen t gia sc ang dn rt khi phm vi s dng, thay vo l ngun collagen c trch t c,
c bit l c da trn: c tra, c ba sa. Ngun collagen c chit tch t ngun nguyn liu ny c nhiu
c im tt hn cho vic ng dng trong thc phm, y hc v m phm. Trong cng ngh sn xut m
phm, m phm trn nn collagen t gia sc c tc hp th rt chm trn da ngi. i mt vi vn
ny, liu php tim collagen vo da c gii thiu. Tuy nhin, nhng bt li ca cch thc ny l t
tin, au n v i km vi n l nhng ri ro nguy him cho con ngi. Hin nay,

cc nh khoa hc Phn Lan pht trin mt tin trnh trch ly collagen t c. Khng ging vi nhng loi collagen thu c
t nhng phng php trc y, collagen t c c hp th hon ton trn da ngi. V c sng trong phm vi rng ln vi
cc iu kin v nhit , su v p sut khc nhau. iu ny c ngha l collagen trch t da c c mt sc chng chu c
bit vi cc ph hy l v ha hc.
i vi cc thuc hay thc ung c cha collagen, collagen t c cng c chng minh l c tc hp th vo
mu nhanh hn gp 1,5 ln so vi collagen t ln.
So snh kh nng hp th vo mu gia collagen t c v collagen t ln

Thi gian sau khi s dng
Hnh . 1 th so snh kh nng hp th vo mu ca collagen trch ly t c v t ln Vit Nam l
mt nc ang pht trin, c nn nng nghip vi cc sn phm phong ph. Trong , cc sn phm t c
chim mt phn ln trong kim ngch xut khu. Nhng nm gn y, vic nui c da trn : c tra, c ba
sa rt pht trin, c bit ng bng sng Cu Long. Trong qu trnh ch bin c, song song vi nhng
chnh phm dng cho
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vic xut khu (ch yu l fillet) l mt khi lng ln cc ph phm nh da, xng, vy,... y l nhng
ngun giu collagen, c th s dng thay th cho nhng ngun cung cp Collagen c in nh da, xng
t ln, b. Vic tn dng nhng ngun ny s gp phn lm tng gi tr kinh t ca cc loi c da trn
nc ta.
1.2 CU TAO COLLAGEN
1.2.1 1.2.1 Thnh phn ha hoc ca Collagen [1]
Thnh phn protein trong collagen c gn y cc loi acid amin, bao gm tt c 20
loi acid amin ( Schrieber v Gareis,2007). Thnh phn acid amin c th thay i ty theo ngun gc ca
collagen, nhng vn tn ti mt vi tnh cht chung v duy nht cho tt c collagen.
Trong thnh phn collagen khng cha cystein v trytophan, nhng cha mt lng ln
glycine (Gly), chim khong 33% v proline (Pro), chim t l 12% v hydroxyproline (Hyp), chim t l
22% (Theo Balian v Bowes 1977). Collagen l mt trong s" t nhng protein c cha hydroxylysine
(Hyl), ngoi ra trong thnh phn Collagen cn cha khong, chim t l 1%.
Bng 1.2 So snh thnh phn cc acid amin trong collagen v cc loi protein khc

(s" gc/1000 gc)

Collagen Casein Albumin
Glycine 363 30 19
Alanin 107 43 35
Valine 29 54 28
Leucine 28 60 32
Isoleucine 15 49 25
Serin 32 60 36
Threonine 19 41 16
Cystein
-
2 1
Methionine 5 17 16
Aspatic acid 47 63 32
Glutamic acid 77 153 52
Lycine 31 61 20

Proline v Hydroxyproline lin quan ti cu trc bc 2 ca collagen. Nhng amino acid ny
gip gii hn s quay ca b khung polypeptide, do gp phn to nn s bn vng cho cu trc xon
c bc 3. Nhm hydroxyl ca hydroxylproline ng vai tr quan trng trong s bn vng cu trc xon c
bc 3 ca collagen. Polypeptide ca collagen m thiu hydroxylproline s to nn cu trc gp khc
nhit thp v s khng bn vng nhit thn nhit.
Hydroxyproline l mt acid amin c trng ca collagen m cc loi protein khc
khng c. Hydroxyproline c hnh thnh sau qu trnh iu chnh proline khi c kt hp vo v tr Y
trong chui Gly-X-Y. Hydroxyproline c cng thc phn t l C5H9NO3, khc vi proline, trong cng thc
cu to ca Hyp c gn nhm OH nm v tr cacbon gamma, acid amin ny c vai tr quan trng trong
s n nh cu trc ca collagen, l dn xut ca proline trong qu trnh hnh thnh chui collagen c s
xc tc ca enzyme hydroxylase proline v s c mt ca Vitamin c gip b sung thm Oxy, thiu
vitamin c s lm chm qu trnh tng hp hydroxyproline, nh hng n qu trnh xy dng cu trc
protein gy ra nhng ri lon trong c th.
Hydroxylycine 7
- -
Arginine 49 25 15
Histidine 5 19 7
Phennylalanine 15 28 21
Tyrosine 5 45 9
Trytophan
-
8 3
Proline 131 65 14
Hydroxyproline 107
- -

o
H
O
HO
Hnh 1.2 Cng thc cu
to ca
hydroxyproline
Phn t collagen bao gm 3 chui xon li vi nhau to thnh collagen triple- helix to
thnh cu trc 3D- mt m hnh l tng cho cc lin kt hydro gia cc chui ( Te Nijenhuis, 1997). Mi
chui trong triple- helix quay theo chiu kim ng h. Triple-helix di xp x 300nm v chui c khi lng
phn t khong 105 kDa (Papon, Leblon v Meijer, 2007). Triple-helix c n nh bi lin kt hydro ni
gia cc chui. Trong cu trc phn t collagen, do tng tc gia cc mch polypeptide lm cho phn t c
nhng vng k nc v vng phn cc mang in tch s to nn kh nng ho nc lm trng n
collagen. S bin tnh collagen lm cho cc cu ni b tch mt phn hoc hon ton gy nn s tch ri cc
chui do ph hy cc lin kt hydro, chnh iu ny lm mt i cu trc triple-helix. M hnh triple-helice
ca collagen da trn nhng nghin cu v hin tng nhiu x si. Chui nhiu x si th hin trn mc
trung bnh ca ton b phn t collagen , v m hnh c trng th hin l s sp xp lp i lp li ca
cc chui polypeptid, thng thng l Gly-Pro- Hyp.
Mt c im c trng ca collagen l s sp xp u n ca cc amino acid trong mi
mt xch ca tng chui xon c collagen ny. Mi si collagen ny c cu to t ba chui polypeptid ni
vi nhau ging nh si dy thng. Cu trc ca collagen tng t nh hnh xon c, mi mt chui
polypeptid trong collagen c cu to t cc acid amin theo mt trt t, thng thng l Gly - Pro - Y,
hoc Gly - X - Hyp.

Hnh 1.3 Trnh t sp xp cc acid amin trong phn t Collagen Trong X, Y l nhng
n v aminoacid khc nhau, c th l proline (Pro) v hydroxyproline ( Hyp) chim khong 1/6 chiu di
chui. Glycine chim gn 1/3 trong tng s" cc amino acid v n c phn b" mt cch u n ti v
tr mi 1/3 xuyn sut trong phn t collagen. Do glycine c cc nhnh ph nh nht nn s lp li ca n
cho php cc chui polypeptide kt hp cht ch vi nhau hnh thnh nn mt ng xon c vi khong
trng nh phn li. Tng cng ch ring proline, hydroxyproline v glycine chim V chui Collagen.
Cc acid amin cn li chim /2

Collagen cng cha cc n v carbonhydrate, c monosaccharide galactose
hoc disaccharide glucosylgalactose, lin kt vi phn d hydroxylysine thng qua nhm hydroxyl chc
nng ca amino acid. T l ca mono/disaccharide ph thuc vo loi collagen v tnh trng sinh hc. Chc
nng ca cc n v carbohydrate vn cha c hiu bit y , nhng nh hng ca chng trong vic

Bng 1.3 S phn b" cc amino acid trong chui polypeptide
Triple helix T l so vi tng b ba(%)
Gly - X - X 44
Gly-X-Y 20
Gly - Y - X 27
Gly - Y - Y 9

gn kt phn bn phn t collagen hnh thnh nn si v bn knh ca si ang c nghin cu.
Ngoi ra, trong phn t collagen c nhng vng gm khong 9 -- 26 amino acid ti cc
im u mt amino hay carboxyl ca chui. Cc vng ny khng sp nhp vi cu trc xon c v c
gi l telopeptide.
1.2.2 C^u trc phn t Collagen [1]
Phn t collagen l mt protein hnh tr, c cu trc bc 4, do cc n v
tropocollagen cu trc bc 3 t hp theo cc hng dc v ngang lm collagen c nhiu mc cu trc.. n
v c bn ca collagen l Tropocollagen, lin kt vi nhau to thnh nhng si nh. Tropocollagen l mt
cu trc dng si hnh ng chiu di khong 300nm, ng knh 1.5nm, gm ba si polypeptide (cn gi l
3 chui a) cun li vi nhau, mi chui a cun thnh ng xon c theo hng t phi sang tri vi 3 gc
trn mt vng xon, hay cn gi l proline I ( xon tri vi 3.3 gc aminoacid/ vng, lin kt peptid c hnh
th cis ), ba chui ny xon li vi nhau theo hng t tri sang phi to thnh ng b ba xon c (triple
helix) hay cn gi l proline II ( xon tri vi 3 gc aminoacid/ vng, lin kt peptid c hnh th trans),
khong cch gia hai gc tnh t hnh chiu ca chng ln cc trc l 0,3 lnm. Hai cu trc ny c th
chuyn i cho nhau v thng th dng II trong mi trng nc th bn hn.


Hnh 1.4 Cu trc ca Tropocollagen ( triple helix )
Cc chui collagen sp xp song song theo chiu dc lin kt vi nhau bng cc lin kt ngang
to thnh cc si theo chu k nht nh. Chng c sp xp so le nhau mt khong 67nm v c 1 khong
trng 40nm gia nhng phn t lin k nhau. Nh vo cu trc c trt t, bn vn c ca cc chui
xon c c chuyn sang cc si collagen, cung cp cho cc m cng, n hi v nhng c tnh c
hc ring. V th collagen rt chc, dai v bn.

1.
5
4
-------------------------------------------------------

300 nm

H
I
(CH
2
)
2
CH
2
---------- C
COOH

*
lys lys
(CH
2
)
2
CH*CH
2
NH3 H3N CH
2
CH
2
(CH. >2

Hnh 1.5 Lin kt ngang gia cc phn t Trpcollagen Cc
tropocollagen s lin kt ngang vi nhau theo kiu cng ha tr vchnh nh
vo lin kt ny m cc phn t tropocollagen hnh thnh nn si collagen. Cc sicollagen
rt div mnh, chng sp xp to thnh mng collagen.
-collagen Procollagen

jpc
Procollagen peptidase

N-terminus end cleaved C-terminus end cleaved
Tropocollagen 3000
Hnh 1.6 Cu trc ca procollagen v s chuyn t procollagen sang tropocollagen
Cc bc hnh thnh nn collagen:
Bc 1: Tng hp chui alpha, giai don trc procollagen trn riboxom v trong mng li ni
cht.
phn ng


u N

Bc 2: S t hp 3 chui alpha hnh thnh procollagen.
- S hnh thnh cu ni disulphide gia cc phn ca chui polypeptide ti u C-
- Ba chui alpha lin kt li, hnh thnh chui xon c, to procollagen.
Bc 3: S giu cc phn t procollagen bng cch hnh
thnh lp mng thm bao bc.
Bc 4: S phn ct cc peptide nh danh ti lp
mng bao bc, kt qu l hnh thnh nn phn t
collagen.
Bc 5: Qu trnh t sp xp ca collagen phn t hnh thnh nn si collagen. Bc 6:
To cc lin kt ngang gia cc phn t collagen ng cnh nhau.

--U
A
1 :
:

Hnh . 7 S hnh thnh
chui xon c ni phn t v gia cc chui polypeptide
Mi chui polypeptide c khi lng phn t khong 100 kDa, to nn tng khi lng phn
t ca collagen khong 300 kDa. Chui a c cu to bi khong 1000 amino acid. Cc chui a khc nhau
(ai, 0,2 v 003) thnh phn amino acid. S phn b" ca cc chui ai, C2 v 0t3 trong cc phn t collagen
khc nhau ty thuc vo s khc nhau v gene.
1.2.3 Cu trc si ca collagen [3]
Phn ln collagen trong mng li ngoi bo c tm thy dng si, bao gm nhng
si mnh, nh. Thng qua qu trnh to si, cc phn t collagen t hp vi nhau hnh thnh nn cc vi si
(microfibril) bao gm t 4 -- 8 phn t collagen hoc vi s" lng nhiu hn s to thnh cc si (fibril).
Nhng si ny c ng knh t 10 4- 500 nm ty thuc vo loi m v giai on pht trin. Cc si
collagen s thit lp nn cc si ln hn (fiber) v cao hn na l cc b si (fiber bundle).

intermolecular trp>le-hielix formation

Hnh 1.8 Cu trc si ca collagen
> Si collagen l cc t hp c cu trc bn kt tinh ca cc phn t collagen. Cc vi si kt
hp to nn si ln hn. Cc si ln hn s kt hp vi nhau to thnh nhng b si collagen. Cc si
collagen c sp xp vi nhng cch thc kt hp v mc tp trung khc nhau trong cc m khc nhau
cung cp cc c tnh khc nhau ca m.
Cc chui collagen sp xp song song theo chiu dc to thnh cc si vi tnh chu k
nht nh. Chng c sp xp so le nhau mt khong 67 nm v c mt khong trng khong 40 nm gia
nhng phn t lin k nhau.
Nh vo cu trc c th bc, bn vn c ca cc chui xon c c chuyn sang cc
si collagen, to cho cc m c c cng, n hi v nhng c tnh c hc ring bit.
C vi lin kt bn trong mi chui triple helix v mt s" lng bin thin cc lin kt
ngang gia cc chui hnh thnh nn nhng t hp c trt t (nh si collagen). Cc b si ln hn c
to nn nh vo s tr gip ca mt vi loi protein (bao gm nhng loi collagen khc nhau), glycoprotein
v proteolycan hnh thnh cc loi m khc nhau t s sp xp lun phin ca nhng thnh phn trn. S
khng ha tan ca collagen l mt tr ngi cho vic nghin cu n cho ti khi ngi ta pht hin
tropocollagen c th c trch ly t nhng ng vt cn non do cha xy ra lin kt ngang hon ton. Tuy
Phn t collagen
B si


nhin, s pht trin ca k thut hin vi (Electron Microscopy (EM) v Atomic Force Microscopy (AFM))
v s nhiu x tia X cho php cc nh nghin cu thu c nhng hnh nh chi tit v cu trc ca
collagen. Nhng tin b ny c ngha quan trng trong vic hiu c cch thc m collagen tc ng ln
mng li t bo, s xy dng, pht trin, phc hi, thay i ca m trong qu trnh pht trin.

L.-1
Overlap
Gap
D-Period
D
Hnh 1.9 S sp xp ca phn t collagen trong mt si


Hnh 1.10 Mt vi dng sinh hc ca collagen
1.3 PHN LOAI COLLAGEN [4]
Tnh n thi im hin ti, c 42 loi chui polypeptide c nhn dng. Chng c m
ha bi 41 loi gene khc nhau, to thnh 27 loi collagen. H collagen c phn loi thnh nhng phn h
khc nhau da vo s t hp ca cc siu phn t. S phn b" trong cc m ca nhng loi protein khc
nhau th hin tnh a dng v phm vi ng ch , t nhng loi collagen ring bit nhcollagen II, X c
trong sn n nhng loi collagen dng si phn b" rng khp nh collagen v V. Nhng loi collagen
khc nhau thc hin nhng chc nng chuyn dng trong cc m v c phng thc sp xp ring bit ca
cc siu phn t. Mt s loi c cu to t mt tam phn (homotrimer), trong khi nhng loi khc c
cu to t nhiu tam phn. Loi collagen phong ph nht l nhng collagen dng si hnh thnh nhng nn
tng cu trc ca da, gn, xng, sn v nhng loi m khc.



Da theo cu trc v cch t chc sp xp cc phn t trong chui, collagen c phn chia
thnh 7 nhm chnh : collagen dng hnh si, FACIT collagen, collagen dng mng, collagen dng gi
cht, collagen chuyn mng, collagen mng nn v cc loi collagen c chc nng duy nht khc. Nhng
loi collagen ny khc nhau v chiu di ca chui xon c cng nh bn cht v kch c ca nhng phn
khng xon c.

Bng 1.4 Phn loi v v tr phn b" cc loi collagen
Dng collagen Loai cu to phn t Phn b"

I [al(I)]
2
02(1)]
Xng, gn, dy chng,
gic mc,..

II [otl(II)]
3
] Sn, thy tinh th, ...
Collagen dng si TIT [al(TTT)]
3
]
Da,thnh mch mu, li
si ca c m( phi, gan,
lch,...

V al(V), o2(V),o3(V)
Phi, gic mc, xng,...

XI <xl(XI)a2(XI)a3(XI) Sn, thy tinh th,...
Mng nn collagen IV
[al(IV)]
2
a2(IV)] al - a2
Mng nn
Collagen vi si VI al(VI),
ct2(VI),o3(VI)
Sn, phi, thnh mch
mu,
Si neo collagen VIT [ai (V)]
3

Da, vng chuyn tip biu
b - da, nim mc ming,...
Dng mng li lc
gic collagen
VIII [al(VIII)]
2
a2(VIII)] Mng t bo,...
X [o3(X)]
3
Sn,...

IX al(IX)a2(IX)a3(IX)
Sn, dch thy tinh, gic
mc,...
FACIT collagen XII [al(XII)]
3

Mng sn, dy chng,
gan,...

Collagen loi I l ph bin nht v thng trong cc m lin kt nh da, xng, gn.
Collagen loi II hu nh tn ti cc m sn. Collagen loi III li ph thuc rt ln vo tui ca ng
vt, da heo cn tr cha ti 50%, theo thi gian t l ny gim 5-7- 10%. Nhng loi colagen khc ch
hin din vi lng rt nh v ch mt vi c quan c
Hn 90% collagen trong c th l cc collagen loi I, II, III v IV. Nhng bnh tt lin quan
n collagen l do s khuyt tt v gene nh hng n s sinh tng hp, s sp xp cng nh cc qu
trnh khc trong s sn sinh collagen mt cch bnh thng.
1.3.1 Collagen dang si
Collagen dng si c in bao gm cc loi I, II, III, V, XI. Nhng loi collagen ny c kh
nng kt hp to thnh mt t hp cu trc siu phn t c nh hng cao, l chui si so le bc 4

XIV [a1(XTV)]
3

Gan, thnh mch mu,
phi, c vn,...
FACIT collagen XIX [al(XIX)]
3

Sarcoma c vn ca con
ngi

XX [al(XX)]
3

Biu m gic mc, phi
da, xng c sn, gn,...

XXI [al(XXI)]
3
Thnh mch mu,...

XTTT [al(XTIT)]
3

Biu b, nang tc, rut, sn
bo, phi, gan,...
Collagen chuyn XVII [al(XVII)]
3

Vng chuyn tip biu b -
da,...
mng XV [al(XV)]
3

Nguyn bo si, cc t bo
c trn, thn, ty,..

XVI [al(XVI)]
3

Nguyn bo si, phin
keratin,...

XVII [al(XVII)]
3
Phi, gan,...

vi ng knh t 25 - 400nm. Di knh hin vi in t, nhng si ny c xc nh bi mt m hnh c
trng vi mt di chu k khong 70nm ( c gi l chu trnh D) da vo s sp xp so le ca mi n
phn collagen ring bit.
Loai I Collagen loi I l collagen chim thnh phn v c nghin cu nhiu nht. N chim
hn 90% khi lng ca xng v l thnh phn chnh ca m lin kt trong gn, da, dy chng, gic mc,
chim thnh phn rt t trong sn, thy tinh th,...
Cu trc triple_helix ca collagen loi I bao gm hai chui xon al ging ht nhau v mt
chui a2 . Trong c th sinh vt, hu ht chui xon a3 c kt hp vo hn hp c cha loi ITT collagen
( c trong da v li si) hoc loi TV collagen ( c trong xng, gn v gic mc). Trong hu ht cc c
quan c bit l trong gn, collagen loi I c chc nng to nn gin, chc. Trong xng, n tham gia
xy dng cu trc ca xng, m bo kh nng chu ti trng, cng chc ca xng.
Loai n
Collagen loi II l thnh phn c trng v chim hm lng cao trong sn trong sut. Tuy
nhin, hm lng collagen loi II ny ngy cng chim t l ln trong tng s collagen ( khong 80%) khi
c tm thy trong cc c quan nh : thy tinh th, cc biu m gic mc, dy sng, ...cu trc triple_helix
ca collagen loi II c c trng bi ba chui xon al hnh thnh mt phn t homotrimeric tng t
nhau v kch thc v c chc nng sinh hc nh ca collagen loi I.
Nhng si collagen khc trong sn ng vai tr kt hp vi collagen loi T, cng nh loi XI v
IX nhm gii hn ng knh trong khong t 15 - 50nm. Khc vi collagen loi I , chui xon collagen
loi II cha hm lng hydrolysine kh cao, cn bng nng ca proteoglycan - mt thnh phn c trng
dng ngm nc ca sn trong sut.
Loai m
Ging nh loi II, collagen loi II cng c to thnh t 3 chui xon al. Collagen loi III
c phn b" rng ri trong cc m ngoi tr xng. N l thnh phn quan trng ca nhng si li
trong cc m h phi, gan, l lch, ... Phn t Homotrimeric thng kt hp vi collagen loi to
thnh mt thnh phn phong ph trong cc m n

Loai V v XI
Collagen loi V v XI nh l mt phn nhm nh trong collgen dng si,cu trc ca hai loi collagen
ny l mt heterotrimer c trng bi 3 chui xon ai , va a2, a3
khc nhau. iu ng ch l chui xon a3 trong cu trc loi collgen XI c m ha bi cc gen tng
t nh chui al trong cu trc collagen loi T, ch khc kch thc ca glycosylation v hydroxylation. S
kt hp gia s ging v khc nhau ca cu trc hai loi collagen ny xut hin kh nhiu trong cc m.
Theo nhng nghin cu trc y cho thy, collagen loi V l mt hnh thc c th ca
collagen loi I v loi TT trong xng, gic mc v nm xen k trong c bp, gan , phi. Collagen loi XI
cng phn phi rng trong c sn nh loi II. Mt lng ln pha cui u amino non- collagen ca chng
ch c tng hp mt phn v s hp nht ca hai loi collagen to thnh cu trc heterotrimer c
Bng 1.5 S phn b" acid amin ca 3 chui a trong collagen loi I v loi in
Thnh phn Thnh phn theo t l
al ( loi I) a2 ( loi I) al ( loi III)
3-Hydroxyproline 1 2 0
4-Hydroxyproline 108 93 125
Proline 124 113 107
Lysine 26 18 30
Hydroxylysine 9 12 5
Glycine 333 338 350
Cystein 0 0 2
Serine 34 30 39
Alanine 115 102 96
Histidine 3 12 6
Valine 21 35 14
Methionine 7 5 8
Isoleucine 6 14 13
Leucine 19 30 22
Arginine 50 50 46
Phennyalanine 12 12 8
Aspartic acid 42 44 42
Threonine 16 19 13
Glutamic acid 73 68 71
Tyrosine 1 4 3

cho l kim sot kh nng lp rp, sinh trng v ng knh ca chng. K t , cu trc triple helix
c cho l mt mn che trong cc m, chng c t trung tm hn l trn b mt. Do vy, collagen
loi V c chc nng nh l mt cu trc si ct li cng vi loi I v III xung quanh trc trung tm. Tng
t nh m hnh ny, collagen loi XI c cho hnh thc ct li ca nhng si collagen loi TT.
1.3.2 Fibril - Associated Collagens with Interrupted Triple helices :FACIT collagen
Cu trc c bn ca nhng loi collagen ny c c trng bi nhng phn thuc collgen b
gin on bi nhng chui ngn khng xon v nhng phn t trimer ha c lin quan n b mt ca
cc si khc nhau.
Loai IX
Collagen loi TX c phn b" trong sn v thy tinh th nh collagen loi T. Collagen ny
thuc loi heterotrimeric, cu trc bao gm 3 chui xon al, a2 v a3 khc nhau to thnh ba on xon
hai bn hnh cu. Phn t collagen loi IX c t dc theo b mt ca si collagen loi II theo hng
song vi nhau. S tng tc ny c n nh bi lin kt cng ha tr t lysine n u amino ca collagen
loi T.
Loai XII v XIV
Collagen loi XII v loi XIV c cu trc gm 3 chui xon al kt hp vi nhau, cc loi
collagen ny c phn b" ch yu trong xng da, gn phi, thnh mch mu,...cng nh collagen loi
XIX v XX, chc nng ca cc collagen ny vn cha dc tm hiu r.
1.3.3 Collagen vi soi
Collagen loi VI c to thnh t 3 chui xon al, a2 v a3 khc nhau vi cu trc xon m
rng v ko di dng hnh cu. Chui xon a3 gn nh l di gp i cc chui khc v chiu di u amino
v cacboxyl kh ln. Nhng si ban u kt hp bn trong t bo theo hng i song vi nhau, tng cp
chng cho ln nhau, sau sp xp bn phn song song nhau. Trong mng li ngoi bo, chng tp hp
thnh nhng t hp si v hnh thnh mng li vi si c lp trong hu ht cc m lin kt, ngoi tr trong
xng.

1.3.4 Collagen chui ngn
Collagen loi X l thnh phn c trng ca sn trong xng sn, xng sng,..., n c to
thnh t 3 chui xon a3 ging nhau, cu trc mng li hnh lc gic vi u cacboxyl ln v u amino
ngn. Chc nng ca collagen loi X vn cha c tm hiu su, cc nh nghin cu vn ang tho lun v
vai tr ca n trong qu trnh ha xng sn v vi ha.
Collagen loi VT c cu trc ging nh collagen loi X nhng c s phn phi khc bit nhau,
v th m chc nng ca n cng khc. Qu trnh hnh thnh mng li trong collagen ny c thc hin
bng cc ni m t bo hnh thnh nn cu trc hnh lc gic.
1.4 TNH CHT CA COLLAGEN
Collagen l mt protein cu trc. Chnh v vy collagen cng c y cc tnh cht ha l ca
mt protein.
1.4.1 Tc dung vi nc
Collagen khng ha tan trong nc m n ht nc n ra, nguyn nhn l do tng tc
gia cc mch polypeptide lm cho phn t c nhng vng k nc v vng phn cc mang in tch s to
nn kh nng ho nc lm trng n collagen. C lOOg collagen kh c th ht c khong 200g nc.
Collagen kt hp vi nc n ra trong nc, dy tng ln chng 25% nhng di tng ln khng ng
k, tng th tch ca phn t collagen tng ln 2-3 ln.
Do nc phn cc tc dng ln lin kt hydro trong lin kt phi tr ca collagen
lm gim tnh vng chc ca si collagen t 3 - 4 ln. Khi nhit tng ln cao, tnh hot ng ca mch
polypeptide tng mnh, lm cho mch b yu v bt u t thnh nhng mch polypeptide tng i nh.
Khi nhit tng ln trong khong 60 - 65c collagen ht nc b phn gii. Nhit phn gii ca
collagen trong nguyn liu cha x l tng cao. Khi nguyn liu kh ht cht khong, th nhit
phn gii ca nguyn liu s gim xung.
1.4.2 Tc dung vi acid v kim
Collagen c th tc dng vi acid v kim, do trn mch ca collagen c gc carboxyl v amin.
Hai gc ny quyt nh hai tnh cht ca n. Trong iu kin c acid tn ti, ion ca n tc dng vi gc

amin, in tch trn carboxyl b c ch (hnh thnh acid yu c ion ha thp). Tri li gc amin b ion ha
to NH
3
+
.
Mi trng H
+

I r
-
I I
COOH NH
3
+
...C1' COOH NH
3
+
...C1'
NH
3
+
...Cr NH
3
+
...C COOH

Mi trng OH

NH
2
COO' NH
2
COO'
____ J_ ____________ _____________ I ____________ 'L
Trong iu kin c nc, nc c th tc dng vi nhm gc c mang in trong kt cu
protidc v nhng ion Na
+
, cr hnh thnh tc dng hp nc ph ca collagen, khin collagen trong mi
trng acid, kim c ht nc cao hn trong nc nguyn cht. Ngoi ra acid v kim c th lm cho
collagen bin i nh sau:

ct t mch mui (lin kt gia -NH3
+
...COO) lm t mch peptide trong mch chnh.
Lm t lin kt hydrogen gia gc -CO...NH- ca mch xung quanh n.
Lm acid amin b phn hy gii phng amoniac.
Cng vi nhng bin i , im ng in ca collagen h xung thp, (v nhng bin i mang tnh
cht thy phn lm cho cc nhm gc c tnh hot ng tng ln
NH,
COO'
...Na
+

COO'
...Na
4

NH
2

HO' H
cr
H
+
OH
HO' H
Na
Na
+
OH H
+

cr
H
+
OH'
H
+
OH'

nhiu).
Acid v kim u c th phn hy Collagen, nhng s phn hy ca Collagen cho ti nay vn cha
c khi nim chnh xc. biu th s bin i chnh xc ca Collagen di tc dng ca acid, kim ni
chung u dng keo phn gii lm ch tiu. keo phn gii l ch s' keo sinh thnh sn phm c th tan
trong nc (thng l trong iu kin un nng), nhng sn phm ny di nng tng ng tin hnh lm
lnh c th bin thnh keo.
Theo kt qu nghin cu tc dng ca vi vi Collagen c th lm tng phn gii ca n rt
mnh, trong iu kin nhit thng thi gian di c th xc tin s phn gii ca Collagen (ty phn
gii ca n rt thp) lc nhit tng ln hoc x l bng kim th phn gii tng ln.
Collagen phn gii bin thnh gelatin, cn c vo gi thuyt ca Hofmeister phn ng s tin hnh
nh sau:
t ;xt
C102H149N31O38 + H2O C102H151N31O39
(Collagen) (Gelatin)
Lc x l nhit cao trong nc, keo d tip tc thy phn thnh:
t
C102 H151N31 O39 + 2H2O C55 Hs5 N17O22 + C47 H70O19 + 7N2
(Gelstose) (Gletone)
T thy rng trong qu trnh trch ly gelatin cn thit phi khng ch nhit v thi gian thch
hp m bo cht lng ca gelatin.
Ngoi ra, Collagen trong dung dch mui trung tnh cng vi g"c -COOH v -NH
2
to thnh cc
hp cht mui. Tc dng phn gii Collagen ca NaCl so vi Na
2
S0
4
mnh hn ( do in ly ca NaCl ln
hn). Nhiu thc nghim cho thy rng tc dng ca trypsine i vi Collagen x l ( nh un nng,
ngm acid, kim , mui, tc dng ca pepsin, tc dng c hc,...) th n c tc dng phn gii, vi iu kin
thch hp nht l nhit 37c, pH = 8,1 - 8,2.
1.4.3 S bin tnh
Di tc dng ca cc cht ha hc nh axit, baz, mui, cc dung mi nh ancohol, cc tc

nhn vt lnh khuy trn c hc, nghin, tia cc tm,... cu trc xon bc
ba cacollagen b bin i lm cho cc lin kt hydro, cc lin kt ion b ph v nhng
khng ph v c lin kt peptid, tc l cu trc bc mt vn gi nguyn.
Sau khi b bin tnh Collagen thng c cc tnh cht sau :
ha tan gim do lm l cc nhm k H
2
0 vn bn trong phn t protein. Tc l khi
collagen cha bin tnh, cc acidamin c cc nhm k nc nm bn trong phn t collagen. Khi
b bin tnh, phn t Collagen lc ny ch l trnh t sp xp cc acidamin mch thng, lm cho cc
nhm k nc l ra ngoi nn ha tan c collagen b gim i.
Kh nng gi nc gim, cu trc bc bn gip cho collagen c kh nng
hydrat ha to thnh mng hydrat bao quanh. Khi bin tnh mt i cu trc bc bn th kh nng
hydratha khng cn, v th nn collagengi"nc km.
Hot tnh sinh hc ban u mt i. iu ny l hin nhin v hot tnh sinh hc
ca mt protein c quyt nh bi cc cu trc bc hai, bc ba v bc bn. Khi cu trc ny mt
i, protein ch l cc polypeptid, v khng c hot tnh sinh hc.
Hot tnh sinh hc ban u mt i. iu ny l hin nhin v hot tnh sinh hc ca mt
protein c quyt nh bi cc cu trc bc hai, bc ba v bc bn. Khi cu trc ny mt i,
protein ch l cc polypeptid, v khng c hot tnh sinh hc.
Tng nhy i vi s tn cng ca enzyme protease do lm xut hin cc lin kt peptide
ng vi trung tm hot ng ca protease.
Tng nht hin ti.
*> Mt kh nng kt tinh, collagen c kh nng kt tinh trong cc mi trng mui hoc cn. cc iu kin
pH nht nh dung dch collagen khng bn v kt
tinh gi l pH ng in. Khi b bin tnh im ng in ca protein khng cn v chng kh c
th kt tinh.
1.4.4 Tnh ky nc
Do cc gc k nc ca acidamin trong chui polypeptid ca collagen hng ra ngoi, cc gc ny

lin kt vi nhau to lin kt k H
2
0. k nc c th gii thch l do cc acidamin c cha gc R-khng
phn cc nn n khng c kh nng tc dng vi nc. Tnh k nc s nh hng rt nhiu n tnh tan ca
collagen.
1.4.5 Tnh cht ca dung dich keo
Khi ha tan protein thnh dung dch keo th n khng i qua mng bn thm. Hai yu t" m bo
bn ca dung dch keo l :
> S tch in cng du ca cc protein.
> Lp v hydrat bao quanh phn t protein.
Khi dung dch keo khng bn, s c hai dng kt ta : kt ta thun nghch v kt ta khng thun
nghch.
> Kt ta thun nghch: sau khi chng ta loi b cc yu t" gy kt ta th protein vn c th tr
li trng thi dung dch keo bn nh ban u.
> kt ta khng thun nghch: sau khi chng ta loi b cc yu t' gy kt ta th protein khng
tr v trng thi dung dch keo bn vng nh trc na.
Dung dch collagen c nht cao trong khong pH = 2 -5 , trong pH = 2 cho nht cao nht,
iu ny gip cho vic nghin cu, tch chit collagen.
1.4.6 Tnh cht Hydrat ha ca collagen
Trong qu trnh hydrat ha, protein tng tc vi nc qua cc n peptid hoc cc gc R mch
bn nh lin kt hydro. Nng protein, pH, nhit , thi gian, lc ion, s c mt ca cc thnh phn khc
l nhng yu t nh hng n cc phn ng protein - protein v protein - nc. Khi nng protein tng th
lng nc hp th tng s" tng, pH thay i dn n thay i mc ion ha v s tch in trn b mt
phn t Collagen, lm thay i lc ht v lc y gia cc phn t ny v kh nng lin kt vi nc.
Khi nhit tng th kh nng gi nc ca Collagen gim do lm gim cc lin kt hydro. Ngoi
ra, bn cht v nng cc ion gy nh hng n lc ion trong mi trng v s phn b" in tch trn b
mt phn t Collagen nn cng nh hng n kh nng hydrat ha. Ngi ta nhn thy c s cnh tranh ca
phn ng gia nc, mui v cc nhm ngoi ca acid amin. Khi nng mui thp, tnh hydrat ha ca

protein c th tng do s nh thm cc ion gip m rng mng li protein. Tuy nhin khi nng mui cao,
cc phn ng mui - nc tr nn ni tri hn, lm gim lin kt protein - nc v protein b sy kh
1.5 CC PHNG PHP NH G V PHN TCH COLLAGEN
C nhiu phng php nh gi v phn tch collagen : phn tch nht, m, hm lng tro,
xc nh khi lng phn t, vi sinh, phn tch thnh phn protein trong collagen (Xc nh tng nit bng
phng php Kjeldahl), phn tch thnh phn axit amin.
1.5.1 Phng php in di trn gel SDS-polvacrvlamde (SDS-PAGE) xc inh trong lng
phn t collagen.
in di theo phng php SDS- PAGE ca phng php ca Kittiphattanabawon, Benjakul,
Visessanguan, Nagai, Tanaka, (2005), s dng Buffer System l my in di ng, loi mini- PROTEAN
Tetra cell ca hng BIORAD.
Mu collagen c ha tan trong dung dch acid acetic 0,5M n nng 2,5 mg/ml. Chnh pH
ca dung dch collagen bng sodium phosphate 0,2 M n gi tr pH 4,4. Sau , ha dung dch mu ny vo
hn hp cha 1% SDS (sodium dodecyl sunfate) v urea 3,5M (pH 7,2), khuy trn lin tc nhit phng.
Hn hp c ly tm loi b tp cht khng tan. Dung dch ny c trn theo t l 1/1 (v/v) vi dung dch
m (0,5M tris-HCl, pH 6,8 cha 4% SDS, 20% glycerol).
To ging: Chng ta pha resolving gel 7% v stacking gel 5% sau ln lt bm vo ging v
ci lc to cc ming ging.
Dung dch mu c bm vo cc ming ging ca my in di, dung dch m vo b, chnh
dng in khong 20 mA/gel. Sau khi in di, gel c nhum mu bng dung dch cha Co-omassie blue R-
250 0,05% (w/v) trong methanol 15% (v/v) v acetic 5% (v/v), trong 1 - 2 gi v ty mu bng dung dch
methanol 30% (v/v) v acetic 5% (v/v) cho n khi xut hin cc vch protein . Protein chun c in di
so snh c trng lng phn t t 75 - 250 kDa.

1.5.2 Xc inh S bin tnh ca collagen theo nhit bng phng php bin i nht
(Denaturaton temperature determned bv vscostv method).
nht ca dung dch collagen c xc nh theo phng php ca L.s. Senaratne v cng s
(2006)
Thng qua vic o nht ca dung dch collagen, chng ta xc nh nhit bin tnh ca
collagen. Nhit bin tnh ca collagen l nhit ti gi tr nht gim i mt na.
Dung dch collagen 0,03 % trong acetic acid 0,1N, c khuy u cho tan ht. Tin hnh o
nht trong mt b iu nhit. Nhit c gia tng rt chm khong lc/ph. Bt u o 33c v kt thc
50c. Mt pht ly gi tr mt ln.
nht ca dung dch collagen c xc nh tng ng cc mc nhit . nht tng i
c tnh nh sau:
ux*. 4. ____
^min
nht tng oi = ----------- -
^lmax
^Imin
Trong :
h
t
: nht o c ti mi mc nhit
hmax' nht ln nht
h
min
: nht nh nht
ng cong bin tnh nhit c xy dng bng cch v th nht tng i theo nhit .
Nhit bin tnh chnh l nhit m ti nht tng i gim i mt

chanr cail sh pig

Tcmr*5tivne (*C)
Hnh 1.11 ng cong bin tnh nhit ca 2 loi collagen thu c t da c v da ln trong cixit axetic 0, N
v nng collagen l 0,03%
1.5.3 Phn tch hm lng hvdroxyprolne
Hydroxyproline l mt acid amin c trng ca collagen .Chng khng c trong bt k mt
loi protein no khc, ngoi tr mt lng nh trong elastin. Do , c th nh lng collagen thng qua nh
lng hydroxyproline v nhn vi h s" chuyn i 14,7 (Sadowska, Kolodziejska, & Niecikowska, 2003).
Quy trnh xc nh hydroxyproline c thc hin theo phng php ca Tgnateva, Danilov,
Averkiev, Obrezkova, Lunin, Sobol, 2005.
Cu trc ca hydroxyproline c cha vng pyrrolidine, n c th b oxy ha hnh thnh vng
pyrrole. Vng pyrrole ny tc dng vi tc cht Ehrlich, 4-(N,N- dimethylamino) benzaldehyde to thnh hp
cht quinoid c mu (mu sc ph thuc vo nhm th v thay i t mu cam n mu hoa c). Thc hin
phng php so mu tnh hm lng hydroxyproline.
1.5.4 o ph hng ngoai infrared (IR) spectroscopy.
Ph hng ngoi IR l mt trong nhng k thut ph c s dng rng ri nht, l phng
php o hp th ca mu th cc bc sng khc nhau ca tia hng ngoi. Mc ch chnh ca vic phn
tch ph R l xc nh cc nhm chc ha hc c trong mu th. Nhng nhm chc khc nhau s hp th


nhng bc sng ring bit. Vi nhiu thit b ph tng, my ph hng ngoi c th lm vic vi nhiu dng
mu t kh, lng n rn. Phn tch ph IR l mt cng c ph bin v quan trng lm sng t cu trc v
xc nh cc hp cht.

Hnh . 12 Ph hng ngoi collagen trong dung dch axit
Collagen trn ph hng ngoi c th thy trn hnh. Nhm amide A pht hin peak c cng
3330 cm'
1
, nhm amide B c thy peak c cng 3080 em'
1
. Trn quang ph cn thy xut hin s
dao ng ca amide bc 1650 crrf , amide bc T
1545 cm'
1
v amit bc III 1237 cm'
1
, cu trc ca protein v amide bc III chng minh s tn ti ca kt
cu c hnh xon c (Surewicz v Mantsch, 1988, Muyonga, 2004).
CHNG 2: CC PHNG PHP SN XUAT COLLAGEN
Trong cc cng trnh nghin cu, qui trnh thu nhn collagen gm 3 giai on chnh :
Giai on ph v t bo:
- Cc phn t collagen khng c kh nng i qua mng t bo, v vy cn
ph v cu trc t bo chuyn collagen vo dung dch. Cc bin php c s dng c th l cc bin
php c hc nh tin hnh ngin vi bt thy tinh hoc ct thch anh, sau tin hnh ng ha
(homogenizator). Thit b ny c chy thy tinh gn vi motor quay v c th iu chnh tc quay theo
yu cu. Cc t bo nm gia chy thy tinh v thnh c s b ph v.


- S dng cc dung mi hu c nh butanol, acetone kt hp vi cc cht ty ra. Cc
ha cht ny gip cho vic ph v cc bo quan ca t bo.
Giai on tch chit collagen:
- Sau khi ph v cu trc t bo, vic tch chit collagen c tin hnh d dng hn,
collagen c th c chit tch bng acid hoc bng enzyme, qu trnh tch chit collagen c thc hin
trong cc reactor nhit lnh v trong iu kin v trng, kt hp khuy trn lin tc m bo
collagen thu c c cht lng tt.
Giai on tinh sch collagen:
- Trong dch chit th, ngoi collagen cn c cc protein tp, cc lipid, mui khong,
loi cc thnh phn ny cn s dng cc bin php khc nhau.
- loi mui khong thng dng phng php thm tch. Thm tch l s khuych
tn vi phn qua mng vn khng thm i vi nhng cht keo ha tan ( protein, mt s polysaccaride...)
nhng thm i vi cc dung dch tinh th ( cc mui ca hp cht hu c c trng lng phn t
thp,...). loi cc protein tp c th tin hnh kt ta phn on bng mui trung tnh hoc cc dung
mi hu c hay dng cc phng php sc k trao i ion, in di, phng php lc gel,...

2.1 CC CNG TRNH NGHIN cu TRN TH GII
2.1.1 Qui trnh chit tch collagen t da heo [6]

Ngm
Co lng
Ra
Loi bo
Trch ly
Kt ta
Ly tm
Kt ta li

Ly tm


Thuyt minh qui trnh cng ngh :
Da heo sau khi thu nhn s qua qu trnh x l ngm vi dung dch Na2S4, sau tin hnh co
lng ri cho qua qu trnh ra vi nc theo t l H
2
0 : da = 6 : l(w/w). Tin hnh ra ln 1 trong vng 30
pht, sau ra tip ln 2 v 3, mi ln c thc hin trong vng 20 pht, nhit 30c. Qu trnh ra
gip loi bt cc tp cht nh phn tht d, mu, m tha, nhy nht,... chun b cho cc qu trnh tip
theo.
Sau khi tin hnh ra vi nc, da heo s c loi bo bng dung dch isopropyl kt hp vi b
sung cht hot ng b mt khng ion. Tin hnh b sung cht hot ng b mt 2 ln vo dung dch cha
da heo v ngm trong vng 6h, trng hp loi bo bng dung dch isopropyl ta tin hnh trong vng lOh.
Da heo sau khi c loi cht bo s c trch ly bng dung dch NaOH 3% (w/v) v
monomethylamine 1,9%(v/v) tch chit collagen, qu trnh c tin hnh nhit 20c trong vng 1
tun m bo collagen c cht lng tt.
kt ta collagen va c trch ly ta dng dung dch acid HC1 5M. n y collagen th
c kt ta, pH lc ny bng 4,6 - 4,7. thu c collagen sch ta tin hnh ly tm ly phn ta ha tan
li trong dung dch dch acid acetic 0.1M, qu trnh c thc hin 4c trong vng 72h, tin hnh ly tm
ln na ta thu c collagen thnh phm.
Nhng kt qu phn tch collagen trn da heo :
Kt qu cho thy collagen c thu nhn t da heo c cu trc ch yu gm hai chui al v a2.
ng vi mi chui c 8 peptide khc nhau v nhng peptide ny c phn tch gia s kt hp trao i ion
v sc k lc gel xc nh thnh phn cc acid amin c trong mi chui. Kt qu cng cho thy chui
collagen c c trng bi v tr v trng lng phn t ca cc acid amin.
Bng cc phng php xc nh cho thy collagen t da heo c nhit bin tnh 37c, nhit
bicn tnh ca collagcn c licn quan cn cc thnh phn acid amin, c bit l

Trch ly
cc thnh phn proline, hydroxyproline, glycine. Nhit bin tnh ny cn ph thuc vo iu kin nhit
mi trng sng ng vt.
2.1.2 Qui trinh chit tch collagen t gn chn g [7]

Ct nh

Kt tinh

Loi protein ha tan
v carbonhydrat

Lc


Nguyn liu l gn chn g sau khi c ct nh c ngm vi dung dch CH
3
COONa
loi i cc protein ha tan v cc carbonhydrat trong thi gian 3 ngy. Dung dch thu c ta tin hnh
trch ly thu c collagen bng cch b sung dung dch m citrat 0,15M.
Dung dch sau khi trch ly c em i lc loi i cc tp cht rn khc, sau tin hnh
thm tch dung dch va thu c loi i cc mui ra khi hn hp tinh sch collagen.
Thm tch l s khuch tn vi phn qua mng vn khng thm vi nhng cht keo ha tan
( protein, mt s polysaccaride...) nhng thm vi cc dung dch tinh th ( cc mui ca hp cht hu
c c trng lng phn t thp,...). Cc tinh th c th khuch tn t dung dch c nng thp hn
(thng l dung dch ra) vo dung dch keo. Trong khi , cc ion v cc cht phn t nh s chuyn t
dung dch keo vo dung dch c nng thp
Trong qu trnh tch chit v tinh sch protein, loi mui ra khi dung dch protein th dung
dch protein c cho vo cc ti c hiu lm bng nguyn liu bn thm, thng l dng cc ti
Cellophan. Sau t c ti vo bnh cha lng ln, H
2
0 hay lng ln dung dch m c pha long.
y tc gi dng dung dch Na2HPC>4 0,02M. V mng cellophan l mng bn thm c kch
thc l ch cho cc cht c trng lng phn t nh i qua cc dung dch m long. Nh vy, mui s
khuch tn vo dung dch m long ( di chuyn theo hng gim nng ), cn dung dch m long s
di chuyn t dung dch ra vo ti cha protein. Protein l nhng i phn t khng th vt qua ti thm
tch v c gi li trong ti.
Sau khi thm tch , tin hnh ra li v sau kt tinh s thu c collagen thnh
phm.
2.1.3 Qui trnh chit tch Collagen t da mc [7]


Ra sch
Lm kh

Ly tm
Sy lnh


Da mc sau khi c ra sch c lm kh ri cho dung dch NaOH 0,1M vo tin hnh
loi nhng hp cht noncollagenous protein v mu. Sau tin hnh ra v trch ly bng dung dch acid
acetic trong vng 3 ngy .
Hn hp sau khi trch ly s c tin hnh ly tm vi tc 50000 vng / pht trong vng lh
thu ly phn dch, phn dch chit thu c l dch lng, hi snh gi l acid-soluble collagen.
kt tinh collagen, cho NaCl sn vo dch chit n nng 0.8M. n y collagen th
c kt ta. thu c collagen sch, ta tin hnh ly tm vi tc 50000 vng / pht trong vng lh
ri tin hnh sy lnh thu c sn phm collagen dng bt.
2.1.4 Ouv trnh trch lv
collagen t da c tuvt (Baltic
cod) [9]

> Thuyt minh qui trnh cng ngh
Nguyn liu l da c c ct nh ti ng knh khong 3mm bng my nghin, sau tin
hnh khuy trn vi dung dch acid acetic nng 0,1-r 0,5M vi t l 1:6-7- 1:40 nhit lnh trong
vng 2h.
Tin hnh ng nht ha dung dch va khuy trn nhit trong thi gian 4 pht, tc
ng nht l 6000 vng / pht.
Sau dung dch c tip tc khuy trn trong thi gian 24h nhit ri c a qua
my ng nht ha vi thng s" k thut ging nh giai on u.
Sau khi hn hp c ng nht vi nhau ta tin hnh ly tm thu ly phn dch c cha
collagen, qu trnh ly tm c tin hnh nhit lnh trong thi gian 20 pht, tc ly tm t 6000
vng / pht.
Cc qu trnh c tin hnh nhit lnh v trong iu kin v trng trnh da c b
bin tnh nh hng n cht lng collagen.
> Nhng kt qu phn tch collagen trn da c tuvt
Tc gi tnh hm lng collagen thng qua vic phn tch hm lng hydroxyproline v
nhn vi h s' chuyn i t hydroxyproline qua collagen l 14,7. Hm lng collagen trong da c tuyt
l 21,5% (tnh theo trng lng t), v 71,2% (tnh theo trng lng kh).
i vi phng php chit bng acetic cho thy hiu sut chit ph thuc rt
nhiu vo nng acid v t l nguyn liu/dung dch. Cho nn khi kho st qu trnh chit, cn tin hnh
cc nng v t l khc nhau tm mi lin h ca chng nh hng n hiu sut tch chit.
i vi phng php chit bng acid citric, vn chit c collagen nhng hiu sut tch
chit khng cao hn acetic. Tuy nhin tc gi ch dng li vic kho st cc qu trnh tch chit m cha
phn tch c nhng thng s" ca collagen nh cc chui cu trc
AMH CNG NGH THC PHM GVHD: TS. PHAN NGC HA

Acid acetic 0.5M
ca collagen, trng lng phn t, v tr v thnh phn acid amin, nhit bin tnh ca collagen,...
2.1.5 Qui trnh tch chit Collagen t phu phm da, xng, vy ca cc loi c ng ng (
Katsuwonus Pelams), c Pecca Nht Bn ( Lacteolabrax iaponias), c Thu (Samber iaponus). c
Trp Vng( Dentex tumifrons), c Mp ( Heterodonus iaponius).

Chit
Ly tm

Kt ta
Thm tch

Ly tm
Sy thng hoa

Butyl alcohol
Loi tp cht
Loi tp cht

Da, xng, v vy c bo qun 25c n khi s dng. Tt c qu trnh tin x l n tch
chit u tin hnh 4c.
- Mc ch: qu trnh loi tp cht nhm mc ch loi i cc cht non- collagen ( cht
bo, cht mu, cht gy mi tanh,...) chun b cho cc qu trnh tip theo.
- Cc bin i ca nguyn liu: da c c qu trnh ht nc, trng n, lc ny trng lng
ca da c tng ln.
- Thit b v thng s" cng ngh : ln loi tp cht u tin, ta tin hnh x l bng dung
dch NaOH 1 %, sau da c c cho x l tip vi dung dch Butyl alcohol 10% loi tip tp cht.
Sau qu trnh loi tp cht, collagen c tch chit bng dung dch acid acetic 0.5M trong 3
ngy. Qu trnh chit collagen c thc hin trong cc bnh reactor nhit lnh, trong iu kin v
trng, khuy trn lin tc m bo chit c collagen c cht lng tt. Sau qu trnh chit collagen
ta tin hnh ly tm thu ly phn dch, c hai phn dch chit thu c l dung dch lng, hi snh gi l
acid-soluble collagen. Phn b thu c cng c chit ln hai vi dung dch aid acetic 0.5M trong vng
2-3 ngy.
kt tinh collagen, cho NaCl sn vo dch chit n nng 0.9M. n y collagen th
c kt ta. thu c collagen sch, ta tin hnh ly tm, ly phn ta ha tan vo dung dch acid
acetic 0.5M. Tin hnh thm tch thu c collagen sch. Collagen sch c lm lnh v bo qun
nhit lnh.
Trong qu trnh tch chit v tinh sch protein, loi mui ra khi dung dch protein thi dung
dch protein c cho vo cc ti c hiu lm bng nguyn liu bn thm, thng l dng cc ti
cellophan. Sau t c ti vo bnh cha lng ln, H
2
0 hay lng ln dung dch m c pha long (
c th dng m phosphat c pH = 7, nng 0.0IM). V mng cellophan l mng bn thm c kch
thc l ch cho cc cht c trng lng phn t nh i qua cc dung dch m long. Nh vy, mui s
khuch tn vo nc hoc dung dch m long ( di chuyn theo hng gim nng ), cn nc hay
dung dch m long s di chuyn

Hnh 2.1 s lm vic thit b sy thng hoa
t dung dch ra vo ti cha protein. Protein l nhng i phn t khng th vt qua ti thm tch v
c gi li trong ti.
Sau khi tin hnh thm tch ta thc hin ly tm collagen va thu c, sau tin hnh sy
thng hoa thu c sn phm :
- Cc bin i nguyn liu: tch nc khi vt liu bng cch bin nc trong vt
liu thnh , sau bin nc thnh hi nc m khng qua trng thi lng.
- Thit b: H thng sy thng hoa tun hon
* H nhit sn phm sy xung di im ng lnh (-10oC-r-20oC)
v c t trong bnh chn khng c p sut gn vi p sut chn khng tuyt i, khi nc thot ra
khi sn phm sy trng thi rn, tc l thng hoa m.
* M hnh h thng thit b sy thng hoa gm 5 b phn: bnh thng
hoa, bnh ngng ca my lnh, my nn ca my lnh, bnh ngng-ng bng, bm chn khng. Qu trnh
sy thng hoa gm 3 giai on: giai on ng lnh, giai on thng hoa v giai on sy m d. sy
thng hoa c u im gim thi gian sy xung 3 ln, khng nh hng ln n gi tr dinh dng v mi
v sn phm, tuy nhin chi ph nng lng ring cao hn cc phng php khc khong 50-60%.

DUQ dch

Cht ti nhit


Nhng kt qu phn tch collagen trch chit t da c
v hiu sut tch chit: t c hiu sut tng i cao 51,4% (c pecca Nht Bn), 49,8%
(c thu), 50,1% (c mp) tnh trn trng lng kh.
v phn tch cu trc chui collagen :

Hnh 2.2 Sc k in di SDS-PAGE collagen loi I ca da heo v collagen da c. (in di trn 3,5% gel
cha urea 3,5M: (A) Da heo; (B) C Pecca; (C) C mp; (D) C thu
Bng phng php in di nh ni trn, hnh 2.2 cho thy collagen da ca c pecca v c
mp gm 2 chui a khc nhau, l al v a2. Trong , a2 c hm lng rt nh. Nu c a3 tn ti th
cng khng th tch khi al trong iu kin in di ny. Cn i vi c thu, collagen dng a ch gm
chui al m hon ton khng c a2. Trong hnh 2.2 ta cng thy c s tn ti ca chui j3 c 3 loi c.
Trong nghin cu ny, tc gi s dng phng php in di SPS-PAGE theo phng php
ca Weber v Osbom ( 1969) xc nh chui collagen. Trong chui collagen c ha tan vo
Sodium photphat 0.02M ( pH = 7.2) cha 1% Sodium dodecyl sunphat ( SDS) v 3.5 ure. Tin hnh in
di trn 3.5% gel vi m photphate 0.1M (pH = 7.2) cha 1% SDS. Tc gi o nhit bin tnh ca
hn hp collagen 0.03% ha tan vo acid acetic 0.1M ti mt s" nhit . - Nhit bin tnh collagen
ca da c rt thp : 26.5c ( c Pecca), 25.6C(c thu), 25c (c mp), thp hn khong 10 c so vi
collagen ca da c heo ( 37C).
Nhn chung, tc gi a ra phng php tch chit n gin, d thc hin, ch l tch chit
da trn cc ho cht thng dng, khng qu t tin, cc thit b cng thng dng trong phng th

(A.) (B) (O) cr

nghim. Tc gi cng kho st c cu trc chui collagen gm al, a2, v a3. Kho st c nhit
bin tnh ca collagen da c v so snh vi collagen da heo. Tuy nhin cng c mt s" nhc im l
khi s dng butyl alcohol trong qu trnh ra da c loi tp cht, phi qua cng on chng ct thu hi
dung mi. Butyl alcohol khng c khuyn co s dng trong thc phm. Tc gi kho st cu trc
collagen nhng cha xc nh c trng lng phn t ca chng, cha xc nh thnh phn acid amin
trong collagen.
2.1.6 Nm 2005. Kittiphattanabawon. Beniakul. Visessanguan. Nagai. Tanaka, trn c s s
dung cc phng php tch chit collagen theo qui trnh trn c kt hp mt s" thav i v pht
trin thm vic tch chit collagen trn da v xng ca loi c ch vng (Priacanthus taxenus).
Tc gi c nhng nghin cu kh hon thin v y cc phn tch v collagen:
phn tch cc ch tiu v hm lng m, hm lng tro, hm lng cc cht bo,
protein v hm lng hydroxyproline. Trong hm lng cht bo trong da c nguyn liu l 0,98%,
cn trong collagen thnh phm l 0,33% (theo trng lng t), cho thy vic x l da c cha loi sch
trit cc cht bo.
- phn tch thnh phn acid amin trong collagen v xc nh trong glycine l acid
amin chnh chim 30%.
- v sc k in di xc nh chui collagen : tc gi cng c nhn nh ging
nh cc bi bo khc, collagen cha chui a gm ch yu l al v a2 ; cn a3 kh tch ra trong iu kin
in di. Ngoi ra, trong collagen cn c cc chui p v . y chng ta cn lu rng vic xc nh s
tn ti ca cc chui a , p, l quan trng, n cho chng ta bit c mt thng tin l chui dng a l
nhng monomer c trng lng phn t < lOOkDa, chui dng p li l nhng polymer c trng lng
phn t t 100-200 kDa, chui dng cng l nhng polymer nhng li c trng lng phn t > 200
kDa.
- Trng lng phn t collagen nm trong khong 186,5-69,5 kDa.
- Vic xc nh nht ca dung dch collagen: tc gi cho rng khi nhit

cng cao cng lm cho mi lin kt hydro b ph v, nh mi lin kt hydro ny cu trc collagen mi
bn. Trn 40
w
c, collagen b bin i thnh mt hn hp cc dy n, i, v ba cun li mt cch v trt
t (random-coil single, double, triple strand). S ph v cu trc collagen gy ra bi s thay i nhit
v c lin h vi s thay i nht ca dung dch collagen.
Tc gi cn phn tch thm nh hng ca pH ln nht ca dung dch collagen.
Dung dch collagen c nht cao trong khong pH t 2-5. iu ny gip cho chng ta nghin cu tch
chit collagen trong dung dch c pH thch hp, trong pH = 2 cho nht cao nht.
Cn i vi nh hng ca NaCl ln dung dch collagen. nng khong 3% NaCl
trong dung dch acetic 0,5M ca collagen, nht khng thay i. Nhng khi tng nng NaCl ln,
nht gim xung. iu ny c gii thch l khi nng NaCl tng, ha tan ca protein gim do lm
l cc nhm k nc nm bn trong phn t protein.

NaOH
CAcid acetic 0.5M
Chit ln 2
2.1.7 Qui trnh tch chit Collagen trn da ca loi c nc bng acid acetic v enzyme pepsin.

Ly tm

Kt ta
Thm tch

Ly tm
Sy thng hoa


Qu trnh tin x l tp cht vn s dng dung dch NaOH 1% v butyl alcohol 10% . Bn
cnh vic tch chit bng acid acetic, qui trnh cn s dng enzyme pepsine. Da c sau khi ra sch tp cht
s c chit vi dung dch acid acetic 0,5M trong vng 3 ngy. Ly tm ly phn dch cho kt ta vi NaCl
b sung m tris - HC1 (pH = 7,5) thu c collgen th. Cn phn b c chit tip bng hn hp acid
acetic 0,5M v enzyme pepsin 10%(w/v) trong 2 ngy , 4c . im ng in ca enzyme pepsin trong
khong pH = 1, pepsin thy phn mnh cc lin kt peptide ni mch do cc nhm amin ca cc acid amin
thm trong trong phn t protein to thnh, sn phm c tch ra l cc peptide v cc acid amin t do.
Trong mi trng acid, pepsin hot ng thch hp nht trong khong pH = 1.5 - 2 , pepsin c bn ti a
pH = 4-5, vng pH ny, hot lc ca pepsin thp, t pH = 5,6 th hu nh khng c hot tnh protease.
Dung dch pepsin trong H
2
0 dng pepsinogen v c hot ha bi H+ vi pH = 1.5-2, chng tin hnh
phn gii gn 30% mch peptide v thy phn protein thnh polypeptide.
H
+

pep sin ogen -> pep sin
protein > polypeptide
Tin hnh ly tm ly phn dch ny vi dung dch Na
2
HP0
4
0,02M (pH= 7.2) trong 3 ngy.
Sau , dch thm tch c ly tm, ly phn kt ta ha tan vo acid acetic
0, 5M . Kt ta collagen bng NaCl b sung m tris - HC1 (pH = 7,5). Ly tm ly phn ta ha tan vo
dung dch acid acetic 0,5M loi b cc tp cht khng tan trong acid. Tin hnh thm tch loi b acid
acetic, lm kh lnh thu c sn phm collagen.
Kt qu phn tch collagen
i vi phng php s dng enzyme chit collagen, ta nhn thy collagen khng tan hon
ton trong acid nhng tan hon ton trong enzyme pepsin. Hiu sut chit bng enzyme t 44,7% trong khi
chit bng acid acetic ch t 10,7%. Vic b sung
enzyme gip cho s ph v cc mng t bo nhanh v mnh hn. V th collagen d dng c tch ra v
cho hiu sut cao.

Kt qu cng cho thy rng nhit bin tnh ca collagen da c nc l 28c thp hn
collagen da heo. Thnh phn acid amin trong collagen loi c nc gm 19 loi, trong hm lng
glycine chim 30% cho thy c glycine l thnh phn amino acid ch yu trong collagen.

Bng 2.1 Thnh phn a.a ca dung dch collagen da c nc chit bng enzyme pepsine.
Thnh phn acid amin S" gc / 1000 gc
Hydroxyproline 67
Acid aspartic 50
Threonine 25
Serin 48
Glutamic acid 87
Proline 103
Glycine 351
Alanine 106
Cystein 2
Valine 17
Methionine 14
Isoleucine 12
Leucine 23
Tyrosine 4
Phennylalnine 10
Trytophan 0
Lysine 19
Histidine 8
Arginine 54
Tng 1000

2.1.8 Nm 2009. Zhang Liu Li c cc nghin cu trn loi c da trn (Largefin longbabel)
Trong nghin cu ny, qu trnh ty ra c tin hnh 4c, trong iu kin khuy trn. Da
c ngm vi dung dch NaOH 0,1M vi t l 1/20 (w/v) trong vng 24h. Tin hnh thay dung dch mt
ln sau 24h loi bt tp cht non-collagen, cht bo, cht mu v mi tanh. Ra li vi nc cho sch.
Sau tip tc loi bo bng dung dch butyl ancohol 15%(v/v), t l 1h- 20 thc hin 2 ln trong vng
12h. Ra li vi nc lnh cho sch, sau tip tc ty mu vi dung dch H2O2 3%, ra 2 ln trong
vng 24h. Ra li vi nc lnh cho sch.
Sau khi c x l, da c chit vi dung dch acetic 0,5M c khuy trn, t l nguyn liu /
dung dch l 1/15, thc hin trong thi gian 24h nhit lnh 4c, sau dch chit c ly tm lnh.
Phn b c chit tip ln 2 bng dung dch acid acetic trong thi gian 12h. Ly tm dch chit, tt c 2
phn dch chit c thm NaCl n nng 0,7M kt ta collagen. Ly tm ly phn kt ta em tinh
sch.
Lng collagen th c ha tan hon ton vo dung dch acid acetic 0,5M loi b cc tp
cht khng tan trong acid. Dung dch c thm tch vi acid acetic 0,1M trong 3 ngy, thay acid mi
ngy. Phn kt ta c lm lnh kh bng thit b ng kh kh (Labconco Freezone 2,5L, USA) thu
c sn phm collagen sch acid soluble collagen (ASC).
Phn b sau khi chit 2 ln vi acid acetic, em chit tip tc vi dung dch acid acetic 0.5M
cha 15%(w/w) pepsin trong thi gian 30 pht nhit 4c, khuy lin tc. Cho kt ta v tinh sch
nh trn, thu c collagen sch pepsin soluble collagen (PSC).
Ngoi cc phng php phn tch collagen nh tin hnh chy sc k in di, o nht v nhit
bin tnh ca collagen, tc gi cn nghin cu su thm v collagen bng cch o ph hp thu t ngoi
ca dung dch collagen bng quang ph k. Kt qu cho thy rng mc d protein c bc sng hp thu cc
i vng t ngoi gn l 280nm nhng collagen khng c bc sng no trong vng ny. c collagen ASC
v PSC u hp thu bc sng l 233nm. Bn cnh , tc gi cn tin hnh o ph hng ngoi FTR
xc nh bc sng hp thu ca cc amide ca c 2 loi collagen l ASC v PSC.
2.1.9 Qui trnh tch chit Collagen s dung qui trnh ln men ca vi khun Bacillus

Da c
Khuy trn
NaCl

Nui, 37 C(24h)

Chit
Lc

Kt ta

Ly tm

Sy thng hoa

Membrance Thm tch
V - 0.3JLOn J

(48h)
Ra sch Nc

2.2 CC CNG TRNH NGHIN cu VIT NAM
2.2.1 Nm 2008. tc gi L Thi Hng Nhan c nhng nghin cu trn loi c basa (Pansasus
Bocourti) nui tai cc tnh ng bng sng cu Long.
Trong nghin cu ny , qu trnh ty ra da c, tc gi s dng cht hot ng b mt LasNa
kt hp vi NaOH loi cc cht nhy, cht bo. V cng s dng H2O2 ty mu. Cn trong qu
trnh tch chit collagen, phng php c p dng l chit bng acid citric nhng hiu sut thp khong
5,76%. Tc gi cng c mt s phn tch trn sn phm collagen thu c. Trong , nhit bin tnh
collagen c xc nh bng phng php o nht, s dng thit b Brookfield viscometer HDBV- II.
Phn tch nht c thc hin trong mt b nc. Nhit b c ci t t 30c ln 50c vi tc
5c/pht v gi 50c trong 10 pht. Kt qu nht gim khi ta tng nhit

Temperature ( C)
Hnh 2.3 Biu s ph thuc nht theo nhit .
v nghin cu cu trc, tc gi s dng phng php chp electron in t qut scanning
electon microscopy-SEM thy r hn v cu trc b mt collagen v so snh vi b mt ca gelatin.


(a) (b)
Hnh 2.4 Cu trc b mt collagen (a) v gelatin (b)
CHNG 3 : NG DNG CA COLLAGEN
Collagen l mt dng protein cu trc si di v hu ht cc chc nng ca
n khc vi dng dng protein ph bin khc nh enzym. Nhng b collagen hay cn gi l nhng si
collagen l thnh phn chnh ca th nn mng t bo cu to nn hu ht cc m v cu trc bn ngoi ca
t bo, nhng collagen cng c tm thy bn trong ca t bo. Collagen c cng ko t rt ln v l
thnh phn chnh ca bng trong y hc, gn, sn, xng, dy chng v da. Cng vi keratin mm, l nguyn
nhn to ra do dai v ang hi ca da v nu n thoi ha s gy ra np nhn dn n tui gi. N to
nn bn ca thnh mch mu v ng vai tr quan trng trong s pht trin m t bo. N c trong gic
mc, thy tinh th ca mt. Collagen cng c dng trong phu thut thm m v phu thut bng.
Collagen thy phn c th ng vai tr quan trng trong vic iu chnh cn nng, ging nh protein khi
ta dng n c cm gic cung cp nng lng.
Collagen thy phn gi l collagen hydrolysate. Ty theo mc thy phn
m collagen c cc ng dng khc nhau. Collagen hydrolysate c s dng ph bin trong cng ngh thc
phm, dc phm, m phm, v cng nghip nhip nh. Chng c s dng nh cht ng, cht gy lng
trong thc phm. Chng c dng nh cht thay th cao su, keo dn, xi mng, mc in la nhn to, l cht


to kt dnh trong dim qut. Chng c dng trong b lc sng cho n thy ngn,...
3.1 NG DUNG COLLAGEN TRONG CNG NGHIP THC PHAM
Collagen c dng lm v bao xc xch, mng bc ko, lm nguyn liu sn xut mt s
loi thc phm dinh dng, thc phm chc nng. D nhin chng phi tri qua mt qu trnh ch bin
t c nhng tnh cht ng dng mong mun, nh tnh tan chy nhit cao v ng c nhit
thp, lc ny collagen ng vai tr mt cht keo bo v ngn chn s kt tinh ca ng. Chnh v vy m
trong cc ng dng ngi ta thng s dng ch yu l dng thy phn ca collagen, c gi l collagen
hydrolysate.
3.1.1 ng dung collagen trong cng nghip sn xut bnh keo
Cng nghip sn xut bnh ko c l l lnh vc c nhiu ng dng ca collagen hydrolysate
nht tnh cho n hin nay. Collagen hydrolysate c s dng trong cc sn phm bnh ko ngt nh:
winegums, gummy bears, fruit chews, marshmallow v licoride. Chc nng ca collagen
hydrolysate ph thuc vo loi sn phm. Chng hn nh trong marshmallows, collagen ng vai tr lm
bn bt. Trong fruit chews v licoride, collagen tham gia to cu trc, dai cho sn phm,...

Hnh 3.1 Sn phm ko do c v mng l collagen hydrolysate Yu cu ca
collagen hydrolysate trong cng nghi sn xut bnh ko:
Collagen hydrolysate cn phi c ha tan hon ton trc c th pht
huy cc tnh nng cng ngh ca chng. t yu cu ny th cn hydrat ha collagen vi mt lng nc
thch hp kt hp vi khuy o hp l cc phn t c th lin kt li vi nhau. Collagen cn c gia
nhit n 60c c th tan hon ton. N c th ha tan rt nhanh khi c a trc tip vo nc nng
ln hn 90c hoc gia nhit collagen c lm t trong nc lnh, qu trnh ny c thc hin trong
vng t 15 pht n lh. Dung dch ny khng to gel c cho n khi n c lm ngui n nhit thch


hp, gel ny c tnh thun nghch vi nhit , do vy n c th tan chy ra sau khi th gel c to thnh.
Nhit tan chy ca chng thp hn nhit thn nhit ( khong 35 --36
0
C)
Collagen hydrolysate c phn hng v kim tra da vo cng c c
trng bng ch sei Bloom. y l ch s' o c mnh ca collagen hydrolysate. Ch s ny cng cao th
bn gel ca collagen hydrolysate cng ln cng mt nng . i vi cc

Hnh 3.2 Collagen hydrolysate lm cht kt dnh trong thanh ko
ng dng to gel in hnh th collagen hydrolysate c Bloom nm trong khong t 100 -T 280. Collagen
hydrolysate c Bloom cao hay thp c s dng trong cc lnh vc khc nhau. Chng hn nh trong
lnh vc gummy th collagen c Bloom cao v trung bnh c s dng nhiu hn v chng to ra mt
cu trc c trng v ci thin kh nng nh hnh, nhng trong sn xut loi ko chewy th collagen c
Bloom thp (70 -T 140) li c a chung hn vi mc s dng nm trong khong 0,2 -- 2,5%. Nu s
dng collagen c Bloom cao hn th hm lng s dng s thp hn.
nht ca collagen l mt trong nhng yu t" quan trng trong cng ngh
sn xut bnh ko. Chng hn nh trong sn xut cc sn phm gum, nht ca collagen c th tc ng
n qu trnh c nh trong sn phm, c bit l khi collagen c hm lng cao. c bit trong qu trnh
sn xut cc sn phm marshmallow th nht ca collagen ng vai tr quan trng hn trong cc ng
dng khc ca collagen, c th n nh hng n qu trnh nh trn syrup, qu trnh gi cc bt kh, kt
hp vi cc phng php to hnh c s dng. Chng hn nh i vi phng php to hnh s dng
khun tinh bt th nht ca collagen yu cu phi thp khi ko c th chy d dng vo cc khun,
cn i vi phng php p n th nht ca collagen phi cao nh hnh cho sn phm sau khi



Collagen hydrolysate l cht nhy cm vi nhit v acid, c bit l khi c
s kt hp gia hai yu t" ny c th lm hng kh nng to gel ca collagen. Nu qu trnh nu din ra
nhanh th collagen c th c a vo ngay t u cn nu qu trnh nu yu cu thi gian di th
collagen nn a vo qu trnh trn sau cng on nu hn ch phn ng Maillard v phn ng nghch
o ng. Vic thm acid vo nn gi mc ti thiu c th c v nn thm ch vo trc cng on
nh trn ti nhit nh nht c th.

3.1.2 ng dung collagen trong cng nghip sn xut sa v cc sn phm t sa
Collagen hydrolysate c s dng nh mt ph gia to cu trc trong cng nghip sa v
cc sn phm t sa. Chc nng ca chng l:
To cc mi lin kt vi nc: collagen c kh nng trng n to lin kt gp 5 ln
Bng 3.1 ng dng ca collagen hydrolysate trong cng nghip sn xut bnh ko
Loi sn phm Chc nng Bloom nht Lng s dng
Gum
To gel, to cu trc,
xy dng tnh n
hi
100-260.
Thp,trung bnh, cao
2 -10%.
Ko Chewy
To kh, do,
n hi
100-150. Trung bnh, cao 0,5*3%.
Ko
marshmallow
To kh, to gel, tc
nhn bn vng
200-260. Trung bnh, cao 2-5-5%.
Ko nuga
To tnh dai, dn hi
100-150 Trung bnh, cao 0,2--1,5%.
Lp mng ph ngoi To lp mng, to
lin kt
120-150 Trung bnh, cao 0,2 + 1%.

khi lng ca chng vi nc. N gip ngn cn s r nc trong cc sn phm
Cng c cu trc cho cc sn phm sa : collagen khng to phn ng vi cc
thnh phn ca sa v ph thuc vo hm lng s dng, collagen c th lm cht cu trc cream, to lp
gel chc t dch lng. Ngoi ra n cn gip n nh cc bt kh ca cc sn phm t sa c bt nhit
thp v ngn cn s hng cu trc cho sn phm trong sut qu trnh tn tr.
Ging nh cc loi protein khc, collagen c kh nng tao bt tt khi c mt
ca ng v sa. S c mt ca cht bco c xu hng ngn cn s to bt, v vy cn phi s dng thm
cc phng php vt l nh : bm khng kh, kh CO2, kh N2 tng th tch v to bt cho cc sn phm
sa c bt. V chnh collagen cng tham gia vo qu trnh n nh, s bn cht cho cc sn phm sa c bt.
Collagen khng mi do khng nh hng n mi v ca sn phm. N
khng chu s tn cng ca tc nhn oxy ha, c kh nng ngn cn s bin tnh v lu gi c mi tri
cy cho sn phm.
Collgen ng vai tr nh mt cht keo lm nhim v bo v v s ng t ca
sa v casein s hon ton hn v ng nht hn khi c mt ca collagen. ng thi khi c mt mt hm
lng nh ca collagen trong cc sn phm sa th cu trc ca sn phm s tr nn mm mi hn.
Bt collagen hydrolysate s c ha trn vi cc thnh phn khc nh :
ng, cc cht tao hng, mt s" cht n nh khc ri a vo dung dch sa. Hn hp sa ngui ny
c khuy trn, khi collagen s trng n v hp thu mt lng nc gp 10 ln khi lng ca chng,
trong qu trnh thanh trng th collagen s c ha tan. Tuy nhin, khi sa c lc trc qu trnh thanh
trng th tt nht nn a collagen vo di dng dung dch nu khng th qu trnh lc s loi b nhng ht
collagen cn ang trng n. Dung dch collagen cng c th thm vo sa ngui km theo s khuy trn
lin tc. Collagen c th thm vo sa trc khi cy ging m khng nh hng n cc iu kin ln men,
collagen khng phn ng vi cc thnh phn ca sa v khng to kt ta. Collagen l mt loi protein, do
n c kh nng b thy phn bi cc tc nhn nh : nhit , acid, kim, cc tia phng x, enzyme, vi

sinh vt. Nhng nhn chug mnh ca gel b ph hy khi collagen chu tc dng ng thi ca pH (nh
hn 4) v nhit ( ln hn 60C) trong mt khong thi gian ln. Do qu trnh tit trng, thanh
trng acid ca sn phm t sa thng khng nh hng nhiu n tc dng ca collagen trong sn
phm.
Mt s" lnh vc c ng dng collagen hydrolysate trong cng nghip sn xut sa v cc sn
phm t sa:
Yogurt: yogurt l sn phm t sa ph bin trn ton th gii v collagen
hydrolysate c s dng cho sn phm ny vi vai tr ca mt cht n nh gip ngn cn s tch nc t
khi ng. Ngoi ra n cn tham gia vo qu trnh to cu trc kem v cm gic bo trong ming v n c
kh nng tan chy ti nhit thn nhit, s tan chy ny i khi cn mang li hng v tt cho sn phm.
Trong cc sn phm kem : colagen hydrolysate ng vai tr ca mt tc nhn
n nh v n c chc nng iu chnh nht ca hn hp, gip bn vng h nh tng cho ti khi c
s dng, ngn cn s bin tnh trong qu trnh bo qun. Collagen khi kt hp vi cc tc nhn n nh khc
c kh nng to cho sn phm cu trc v hng v thch hp. Bn cnh , collagen gip tng gi tr cm
quan cho sn phm, khi n to cm gic kem, bo cho sn phm m khng to cm gic nht dnh nh cc
sn phm c hm lng bo
Cc sn phm sa ln men c qua qu trnh x l nhit: qu trinh thanh trng
hay tit trng cho cc sn phm sa ln men lm tng thi gian bo qun cho sn phm nhng li lm mt
tnh n nh ca cu trc sn phm dn n hin tng tch nc. Vic b sung hn hp collagen
hydrolysate v tinh bt trc khi thanh trng s gip to cu trc tt v ngn nga c bt k s tch lng
no.
Cc sn phm trng ming c s dng sa nh thnh phn to gel, hng v
cho sn phm : trong cc sn phm ny collagen hydrolysate cng ng vai tr l cht n nh n b ha tan
trong qu trnh x l nhit v s c mt ca collagen hydrolysate gip ngn cn s tng nht v tham gia
to gel cho sn phm khi c lm ngui. N c th c s dng ring hoc kt hp vi cc cht to gel

khc nh carrageenan to nn mt cu trc gel mm mi.
Trong sn phm ph mai, phn ln collagen hydrolysate c b sung vo
ng vai tr ca mt cht nh ha v tc ng vo cc mt lin kt vi nc. Khi so snh cc sn phm ph
mai c s dng thm collagen hydrolysate th mc m, cu trc v hng v ca sn phm tt hn so vi
cc loi ph mai c sn xut theo phng php truyn thng.

Hnh 3.3 Sn phm Ph mai c cha collagen hydrolysate lm gim hm lng bo
> Trong cc sn phm t calo lm t sa, cc sn phm ny i hi phi ct
gim lng cht bo, thm ch l loi khi sn phm cc cht bo t sa. Thay th vo l loi cht bo c
ngun gc t thc vt hoc l kt hp gia cht bo t thc vt v cht bo c sn trong sa. lm bn
vng h nh tng nh th cn phi c s hin din ca cht n
nh v cht nh ha. Cc cht ny gip m bo mi lin kt vi nc, ci thin cu trc( chc,
c, cng) cho sn phm. Collagen hydrolysate c Bloom cao c kh nng thc hin c chc
nng ny v ngoi tnh nng thng thng ca n, loi ny c nhit tan chy xp x bng nhit ca
nhiu loi cht bo thng thng (31 --33
0
C). Mt s" cht keo a nc khc nh : pectin, carrageenan,
xanthan c th kt hp vi collagen hydrolysate tng kh nng lin kt vi nc.
Trong cc sn phm thc ung t sa th collagen hydrolysate c vai tr n nh cu
trc, trnh hin tng tch pha trong thi gian bo qun.
Bng 3.2 Loi v lng collagen hydrolysate s dng trong cng nghip ch bin sa v cc sn
phm t sa.


3.1.3 ng dung collagen trong cng nghp sn xut ung
Cc thc ung nh ru , ru tri cy, cc sn phm nc qu cha ln men
c cha cc thnh phn khng ha tan lm c sn phm thng khng t hiu qu cao nu dng phng
php lc loi b. T thi nn vn minh La M hoc cng c th l vo trc , collagen hydrolysate
c s dng cho qu trnh lc trong ru.
Collagen hydrolysate l tc nhn tinh sch ph bin trong qu trnh sn xut
ru vang . Khi c b sung vo sn phm collagen hydrolysate c tc dng nng cao thi gian bo
qun sn phm, vi kh nng to ta nhanh v to cho ru c mu sng m khng bin i mu c trng
ca sn phm.
Hin nay collagen hydrolysate c s dng ph bin cho qu trnh lm sch
ru vang, bia v nc tri cy v n lm gim c, tng gi tr cm quan cho sn phm m khng nh
hng ti hng v ca sn phm cui cng.
Cc phn ng ca qu trnh tinh sch din ra vi s tham gia ca collagen hydrolysate:
ng dng mnh Bloom Lng s dng
Yogurt 150 0,2 + 2%
Cc sn phm sa ln men qua x
l nhit
150 0,2 + 2%
Jelly hng sa 150 0,2 + 2%
Kem 150 0,2 + 1%
Cc sn phm kem trng ming
150 0,2-2%
Cc sn phm c hm lng calo
thp
250
o

L
/
1

I
-


Phn ng c bn xy ra vi collagen hydrolysate l phn ng to phc gia
polyphenol c trong ru v protein ca collagen hydrolysate to nn cc bng t
lng xung.
Phn ng th hai t c bit n hn nhng khng ng vai tr quan trng l phn
ng to phc gia protein c trong ru v protein thm vo ca collagen hydrolysate.
Cc yu t" nh hng n qu trnh tinh sch c s tham gia ca collagen hydrolysate:
Nhit : iu kin lnh thng thch hp cho qu trnh keo t v lc. Ngi ta chng
minh c rng qu trnh bng t kh xy ra nhit 25 --30
o
C v thm ch l
khng th i vi cc loi ru trng. Lng cht kt ta nhiu thu c nhit
thp c th c gii thch bi s ng
lng ca cc thnh phn hn l s kt hp gia polyphenol vi collagen hydrolysate
trong qu trnh tinh sch.
acid : trong cc sn phm ru, acid cng yu th qu trnh ng t s din ra
nhanh hn, ng thi hm lng collagen hydrolysate s dng cng thp hn.
Bng 3.3 Lng collagen hydrolsate s dng trong qu trnh lc trong cc sn phm ung

3.1.3 ng dung collagen trong cng nghip ch bin thit
Collagen hydrolysate c s dng t nhiu thp nin sn xut ra cc mn
tht ng, lm v bao cc sn phm xc xch. Vo thi im ny collagen hydrolysate cn c s dng
rng ri tim vo tht nhm mc ch tng hm lng protein trong sn phm. V kh nng to lin kt tt
vi nc, collagen hydrolysate cn c s dng tng hm lng m v h gi thnh sn phm. Mt s
sn phm tht c s dng colagen hydrolysate
Gim bng : i vi gim bng co" qu trnh hun khi, bt collagen
hydrolysate hp thu nc trong tht v trong qu trnh ch bin s to mt lp mng gip hn kn khi tht
khi lm ngui. i vi sn phm gim bng c qua qu trnh nu, collagen hydrolysate to gel vi dch lng
tch ra trong qu trnh ch bin, qu trnh ny gip gi nc bn trong v chung quanh sn phm. Collagen
Sn phm Lng s dng
i vi cc sn phm ru

Ru trng 2-=-7g/hl
Ru nho hng 3 + 8g/hl
Ru 12+15g/hl
i vi cc sn phm nc to p

Sn phm lm t to ng 70-120 g/hl
Sn phm lm t to ngt 50^ 100 g/hl
Sn phm lm t to cht 30 + 80 g/hl
Sn phm lm t to chua 20 -r 60 g/hl
Sn phm bia Khong 450g cho 1501 bia

hydrolysate cng gip lm cng chc lp thch thu c trc tip t cc m lin kt nhm to v b ngoi
hp dn v nht ct p cho sn phm. Bcn cnh , collagen hydrolysate c thm vo cc sn phm gim
bng nhm tng hm lng protein.
Tht hp : collagen hydrolysate to gel vi nc tht thot trog qu trnh
ch bin v tranh trng. Collagen hydrolysate c s dng trong sn phm ny l loi collagen c bn
gel cao ( 200 -r 250 Bloom) vi lng 0,5 -r 2%.

vi cc sn phm tht c hm lng nc v hm lng cht bo rt
d xy ra hin tng tch nc, tch bo nh hng n cu trc ca sn phm. Collagen hydrolysate
gip lin kt nc, lm bn h nh tng, to cu trc ng nht. Lng collagen s dng ph thuc vo
s c mt ca cc tc nhn lin kt khc.

Hnh 3.4 Sn phm tht ngui c cha collagen hydrolysate

3.2 NG DUNG TRONG Y HOC V DC PHAM
Collagen l mt vt liu c kh nng phn hy sinh hc, tc dng ca collagen da trn s
kt hp kh nng to mng v to gel mt cch t nhin. Collagen c s dng lnh vc y dc c mi
v v trung ha, khng gy d ng v c hp th hon ton trong c th con ngi, c tnh tng thch
sinh hc cng nh kh nng cm mu nn c th c ch to thnh nhng dng khc nhau, l mt loi
vt liu sinh hc l tng cho vic sn xut cc sn phm y hc.
Cc c tnh k thut ca collagen c ng dng trong cng ngh dc phm :
L mt cht keo bo v cc thnh phn nhy cm khi tc ng ca s oxy ha nh
sng, s hp th m.
To mi v trung ha.
L cht n nh ca h nh tng, huyn ph.
Collagen c hp th d dng vo trong c th v khng gy d ng.
L cht to bt v n nh trong qu trnh sn xut cht bt cm mu.
Bng 3.4 ng dng ca collagen hydrolysate trong cng ngh ch bin tht
Sn phm Chc nng Bloom nht Hm lng
Gim bng To lin kt 200 -r 250 Trung bnh 1 -r 2%
Tht ng
To gel, to cu trc
150 -T 280 Trung bnh - cao 3,5 -r 8%
Tht hp To cu trc 250 4- 280 Trung bnh - cao 1,5-3%
Tht b mui To lin kt 250 -r 280 Trung bnh - cao 1,5 -r 3%
Pate
Tc nhn lm n
nh
180 + 250 Trung bnh - cao 1,5 + 3%
Tht nu chn
ng lnh
To lin kt vi tht
200 4- 240 Trung bnh - cao 0,5 -r 3%

C th s dng lm cht thay th mu tm thi.
Da vo cc c tnh k thut trn collagen c th c ng dng trong cc lnh
vc sau:
3.2.1 Cc ng dung trong qu trnh sn xut thuc
Vin bao nang cng
Collagen l thnh phn chnh trong vin bao nang cng, chng c to ra bng cch
nhng khun kim loi vo dung dch collagen cn ang nng. Collagen s to gel v to nn mt lp mng
lin tc xung quanh khun. Lp mng s c sy kh v ly ra khi khun.
Trong ng dng ny s dng collagen c ngun gc ch yu t xng b, da b v da
heo. Trong nhng nm gn y ngi ta nghin cu tm ra vt liu c ngun gc khng phi l ng
vt lm ra cc vin bao nang. Nguyn nhn chnh ca vic nghin cu ny l vt liu lm vin bao
nang khng c ngun gc t ng vt s p ng nhu cu ca ngi n king v cc thnh phn tn gio
khc nhau.

Vin bao nang mm
Vin nang mm c th c hnh vin bi, hnh trng, hnh tr, hnh git, hnh ht u cha dung
dch lng snh .

Hnh 3.5 Vin bao nang cng

Hnh 3.6 Vin nang mm c lp v ngoi lm t collagen.
Collagen c tc dng bo v thuc chng nhng tc nhn c hi nh nh sng v oxi. Thnh
phn chnh ca v bao l collagen v cc t dc khc nh: glycerol hay sorbitol, nhng cht hot ng b
mt, cht mu cho php, hng liu.
Vin con nhng
Trong vin con nhng collagen c s dng nh vo cc c tnh lin kt ca n. Chng
c s dng nn trc tip sau khi bt thuc c nghin nh. Trong ng dng ny collagen gip to
cho vin thuc cng, kt dnh.
Collagen c ng dng lm mng bao cho cc loi thuc c tc dng c bit, gip chng li
nh hng ca dch v. K thut ny m ng cho vic iu tr s dng chnh xc lng thuc cn dng.
Cc vitamin tan trong du nh vitamin A v E, carotenoid v cc acid bo cha bo ha,... vi
s h tr ca collagen s d dng hn trong vic nh lng, thao tc trong sn xut da vo nht n
nh ca collagen khi kt hp vi cc thnh phn khc trong thuc.

Trang 74

Hnh 3.7 Cc bt collagen c tc dng cm mu SVTH:
V TH HNG LINH
Ngoi ra, collagen cn gip bo v cc thnh phn vitamin trc cc thnh phn nhy cm nh
oxy hoc nh sng. Trong k thut sn xut cc vin thuc si bt nh cc vin tng lc a vitamin th thnh
phn nh ha bao gm collagen, ng, thnh phn nhy cm ca thuc s c sy phun v to lin kt
cho ra cc loi thuc c c tnh mong mun.
3.2.2 Thnh phn cm mu v hn gn vt thng
Cc bt collagen c th c s dng trong qu trnh cm mu, do gip tng
kh nng hn gn vt thng, v vy m collagen c s dng rng ri trong lnh vc nha khoa v phu
thut.
Mt trong nhng thuc tnh gip cho nhng bt ny c th pht huy hot tnh sinh hc mt
cch t u l : khi chng hon ton c ti hp thu trong c th, chng c th c gi li ti cc m th
d nh sau khi phu thut, ti cc m c chng s kch thch cc t bo mi i vo cc vt thng do s
tng tc qu trnh ti sinh ti m c b tn thng. Khi c cy vo cc m c, cc bt ny c th c ti
hp th trong vng khong 3 tun m khng li phn d no. Cn nu chng c a vo ti b mt ca
m c b h hng th chng s hon ton b ha tan trong vng t 3 -T 5 ngy. Cc bt collagen ny c th
ht cht lng ca m c. Cu trc xp cho php n c kh nng ht cht lng c khi lng gp 50 ln khi
lng ca chng



Vt bng mi, trc khi s 3 ngy sau khi s dng
dun collen collcien
10 ngy sau kh s dng collagen
Collagen dng si c dng trong vic lm lnh cc vt thng, vt rch trong phu thut.
Si collagen c th c x l to cu trc si thng, di dng lm nhng sn phm y hc cho gn v
dy chng. Nhng ng collagen (collagen tube) c s dng thay th cho cc cu trc nh thc qun, dy
thn kinh ngoi bin, niu qun. N cng c s dng trong vic nui cy t bo.
Hnh 3.8 Collagen p dng trong vic iu tr vt bng
3.2.3 Tc nhn thay th mu
Trong trng hp mt mu nghim trng, c th ni chung v b no ni ring
s din ra tnh trng thiu oxy. Nu khng c sn mu truyn vo, plasma expander c th c s
dng nh mt gii php tnh th. Plasma expander gip thay th lng mu mt, phc hi li th tch
mu v duy tr mc p sut thch hp. Tuy nhin dung dch ng trng cha cc ion mui v c c khuynh
hng thot ra ngoi m c kh nhanh.

Plasma expander bao gm pyrogene free water, collagen, mui trong collagen chim
khong 3,5 -T 5,5%. Loi collagen c s dng trong trng hp ny yu cu tinh khit rt cao, i khi
chng cn c hiu chnh bng phng php ha hc t c cc thuc tnh v phi tri qua qu trnh
kim duyt kht khe m bo khng gy ra bt k d ng no cho c th

Hnh 3.9 Plasma thay th Collagen dng mng v lp mng c s
dng gi c nh cc vt cht sinh
hc chng hn nh nhn t XT 11 trong mu, dng trong s ti to cc m, ni kt li vng
mc, lm mng thm tch mu, lm vt thay th lp mng cng ca no. N cn c dng
cho vic ti to dy thn kinh, khi phc mng nh, sn v xng, kim sot s chy mu cc
b, khi phc tn thng ca gan, l lp mng chn bo v no, c tc dng phc hi cc vt
thng.
3.2.4 Nui cV m v t bo
S pht trin ca t bo vi s h tr ca collagen trong s nui cy t bo va
c xem xt gn y. Vic s dng cht nn collagen cho s pht trin ca t bo da c cng b" mt
cch rng ri. Nhng t bo nui cy trn collagen kch thch, lm vt thng mau lnh. Nhng nghin cu
rng ri v s pht trin ca cc t bo da t sinh v khc loi trn nn collagen chng minh tnh kh thi
to ra nhiu chng loi m v cc c quan trong qu trnh nui cy. s dng ngun nguyn liu t bo nui


cy thuc nhiu chng loi khc nhau c tc ng to ln n vic iu tr cho cc bnh nhn c nhng tn
thng v m hoc cc c quan.
Nhm cc nh nghin cu trng i hc Wisconsin-Madison to ra dng collagen bn nht
c khoa hc bit n hin nay. y l gii php thay th n nh collagen ngi v mt ngy no c
th c dng iu tr chng vim khp v cc tnh trng thiu collagen. Collagen l dng protein c
nhiu nht trong c th ngi, to ra cc khi v b dy chc chn, c tc dng h tr cu trc ca da, cc
c quan, sn v xng, ng thi l t bo lin kt gia cc b phn ny. Nhiu thp k qua, cc bc s s
dng collagen t b iu tr cc vt bng nghim trng v cc vt thng khc. Collagen nhn to l liu
php cha tr nhiu bnh nh vim khp do thoi ha collagen t nhin trong c th. to ra loi collagen
mi ny, cc nh khoa hc thay th 2/3 lng acid amin thng thng ca protein bng cc dng t do,
gip cu trc chung ca protein cng hn v duy tr c hnh dng ca chng. Tnh t ph ca cch tip
cn ny l s dng cc vt liu cng hn c hnh dng tng t vi cc hnh dng protein chc nng sau khi
gp np. Dng collagen mi gn kt vi nhau nhit cao hn nhit collagen t nhin phn r. Mc d
c to thnh ch yu t cc acid amin khng c trong t nhin, nhng hnh nh chp tia X tinh th cu
trc 3 chiu ca collagen to ra trong phng th nghim cho thy, khng c s khc bit vi collagen t
nhin. Sn phm collagen bn vng thc s l mt bng chng v kh nng ca ha hc protein hin i.
3.2.5 ng dung collagen trong k thut ni soi v phu thut thm m
Trong phu thut ni soi, collagen hydrolysate c ng dng bi vo cc ng ni soi, c
tc dng bi trn, v th cc bc s d dng a cc ng ny vo c th bnh nhn m khng gy au. Sau
thi gian t 40 - 60 pht, collagen s tan hy trong c th bnh nhn m khng gy hi g

Hnh 3.10 ng dng cllagen hydrolysate trong k thut ni soi Collagen c s dng rng ri
trong phu thut thm m, collagen l mt polymer t nhin, n c dng trong phu thut to hnh nh
bm mi, cng da mt.

Hnh 3.11 Collagen dng trong phu thut thm m Ngoi ra, collagen cng c bn
trn th trng nh mt cht b sung, ci thin tnh linh hot ca khp. Ngoi ra, collagen cn l phn nn
gip tch t calcium trong xng. Nu hm lng collagen gim ng k, calcium khng th tch t lm cho
xng gin v d gy, sn d b hao mn to nn cc cn au khp gi v hng. Nh vy, xng cng
cp hn, ngoi calcium cn b sung mt lng ln collagen.


3.3 NG DUNG COLLAGEN TRONG CNG NGH M PHAM
3.3.1 Tc dung ca collagen trong cng ngh m phm
Collagen c tc dng chng lo ha, ngn nga v ci thin np nhn
Khong 70% cu trc ca da l collagen, phn b" ch yu lp h b ca da. Collagen to
ra mt h thng nng , h tr cc c tnh c hc ca da nh sc cng, n hi, duy tr m, lm cho
da c mn mng, ti tn v tr trung. Collagen gip duy tr m ti u cho t bo. Ngoi ra, collagen
cn m bo sc t" da, lm sng mu da. S suy gim v cht lng, s lng collagen s dn n da tr
nn kh, mt cng, n hi v thc y qu trnh lo ha ca c th. Chnh v vy m collagen ng vai
tr quan trng gip ci thin cu trc da, kch thch qu trnh ti to ca ln da, phc hi t bo da b tn
thng.

Hnh 3.12 Vai tr ca collagen i vi da Khi c th lo ha, protein ca t bo kt
dnh vi nhau hoc thay i hnh dng lm cho chng khng cn m nhn c chc nng bnh thng.
Collagen b mng i v tnh n hi gim i r rt. Hu qu l cc np nhn da xut hin. hn ch s
hnh thnh ca
UA uko HA H TN IM KHK MNH
T/l. kim dn hAt, kh Mm, teng, cng mn

Y'u. lng, krm dikn hi Khoe mnh, lun hi
^
m Cht kch ng Mrtanocyt

Age
np nhn da, phng php chng lo ha bo v collagen v tng cng tng hp collagen c xem l hu
hiu.
Khi c th bc qua tui 30, tc tng hp collagen b chm li, tc phn hy gia tng.
Trung bnh mi nm, cu trc collagen b mt i khong 1,5% v khi lng; v th cng ln tui, ln da
cng b lo ha nhanh. Trc tui 45, 1/3 trong s" tng lng collagen ca c th c th b mt i. Cc
np nhn, np gp xut hin ngy cng nhiu trn trn, quanh ming v khe mt. Collagen mt i cng nh
hng ng k n s nh hnh cc ng nt trn khun mt, ln da b chng nho, b chy, iu ny c
bit thy r khi ph n bc qua tui 40. Ngoi ra, c rt nhiu nhn t" gp phn vo s suy gim ca
collagen. Cc cht nhim c cha c t toxin s kch thch qu trnh oxy ha, to ra cc gc t do. Cc
gc t do ny ph hy t bo bng cch ly i mt electron ca mi t bo. Chng cn l nguyn nhn gy
ra cc bnh nghim trng nh ung th. Ht thuc, ch dinh dng khng cn bng, thiu cc nguyn t
vi lng, cc loi thc phm ng gi cha nhiu cht ha hc c hi v khng tp th dc thng xuyn l
mt s' nguyn nhn y mnh s mt mt collagen.

Hnh 3.13 S lo ha da S lo ha t bo c tr hon nh collagen
v hyaluronic acid. Cc cht ny lm phng v lp y cc np nhn ca biu b, xc tin qu trnh ti to t
bo da. Nhng np nhn c v quanh vng mt s gim xung nh vo s ci thin ca biu b.

Age 50+

Phn ln cc sn phm collagen nguyn cht u c chit xut trc tip t da ng vt l ln,
b, cu v c da trn; c bit hn, c mt s loi collagen c chit xut trc tip t c th ngi, chnh
v vy chng hon ton khng c cc cht ha hc hay c t", kh nng tng thch vi c th ngi c th
ln ti 100% m khng h c phn ng o thi hay loi b. Tt c mi ngi mi la tui u c th s
dng v s lnh tnh ca chng. Ngoi ra, collagen cn c a vo da bng ba phng php chnh:
Thoa ln da v nh tc ng ca cc cng ngh xung in hay nh sng dn chng vo
su ti vng da cn iu tr.
Tim trc tip vo da.
n ung.
Ba phng php trn u l nhng bin php rt an ton khng h nh hng ti da v c th.

(a) Trc khi iu tr (b) Sau khi iu tr
Hnh 3.14 Tc dng chng lo ha ca collagen
Collagen c tc dng cao trong qu trnh phc hi v ti to da
iu tr collagen phc hi: thng c s dng trong trng hp da b tn thng hay trong
giai on ti to sau khi iu tr nm, mn trng c, so, rn da v tiu da tha sau khi gim bo. Cc sn
phm vi thnh phn chnh l collagen, elastin,... trong collagen c chit xut t da heo gip kch thch
t bo v sa cha cc khim khuyt ca m nh phc hi kh nng n hi ca da. Ngoi ra, sn phm
cn c th iu tr cc vt rn do tng cn hay mang thai.
iu tr tr ha bng collagen 25% c s dng cho ph n t 35 tui bt u c s lo ha


vi vic xut hin cc np nhn, da b mt nc, chng nho, chy x, khng cn cng mn nh trc, mu
sc da cng tr nn sm li. Collagen thc y qu trnh ti to m nhng vng da b so v mn. Mt
trong nhng chc nng ca collagen l to ra mt mng li gi cho t bo da c nguyn vn v gip
chng di chuyn ra b mt ca da. Khi cc t bo mi ny di chuyn ra bn ngoi, chng s in y cc l
su ca m da, v lm gim s xut hin ca so. Collagen cng c tc dng cng c tnh nguyn vn ca t
bo da bng cch kt hp vi cc m lin kt trong da ci thin cu trc v tnh n hi.

Hnh 3.15 Tc dng tr so mn ca collagen Collagen thc y qu trnh ti to m ti
nhng vng da b so v mn. Mt trong nhng chc nng ca collagen l to ra mt mng li gi cho t
bo da c nguyn vn v gip chng di chuyn ra b mt ca da.
Khi cc t bo mi ny di chuyn ra bn ngoi, chng s in y cc l su ca m da, lm
gim s xut hin ca so. Collagen cng gip cng c tnh nguyn vn ca t bo da bng cch kt hp vi
cc m lin kt trong da ci thin cu trc v tnh n hi.
3.3.2 Cc ng dung ca collagen trong cng ngh m phm
Collagen cha cc tc nhn lm m t nhin i vi c th, c vai tr chng nhn, gi cho da
mm v sng; hn na colagen cn dng lm nguyn liu sn xut cc sn phm nh: mt n, kem dng
da cao cp, du gi, cc sn phm dng tc cng nh cc loi sa tm...
Collagen hydrolysate l thnh phn trong cc sn phm mt n dng da, chng kt hp vi
cc tinh cht trong mt ong, du oliu, tinh du hoa hng... c tc dng dng m cho da, ti to ln da mt
mi, bo v da khi cc tc nhn tia t ngoi, khi bi, ha cht... V th da c tr ha, ti tn.
Acne Care (4 week program)

(a) Trc khi iu tr (b) Sau khi iu tr
Atmt ntTAr rmarmrtnr

Hnh 3.16 Mt n dng da collagen hydrolysate
Trong thnh phn ca mt n dng da, collagen c b sung da trn nhng cht nn. l
cc hp cht carbomer, acrylate polymer. Cht nn ny c pha v tin hnh chnh pH ln gi tr khong
pH = 7 v gi tr ny s cho ta dung dch cht nn c v trong nht. Collagen trc khi cho vo dung dch
cht nn phi c pha vo nc cho trng n, sau s c trn chung vo vi dung dch cht nn c
sn, t l collagen cho vo chim t l 0,3 -T 0,6%.
Trong cng ngh sn xut m phm, m phm trn nn collagen t gia sc c tc hp th
rt chm trn da ngi. i mt vi vn ny, liu php tim collagen vo da c gii thiu. Tuy nhin,
nhng bt li ca cch thc ny l t tin, au n v i km vi n l nhng ri ro nguy him cho con
ngi. Hin nay, cc nh khoa hc Phn Lan pht trin mt tin trnh trch ly collagen t c. Khng
ging vi nhng loi collagen thu c t nhng phng php trc y, collagen t c c hp th hon
ton trn da ngi. V c sng trong phm vi rng ln vi cc iu kin v nhit , su v p sut khc
nhau. iu ny c ngha l collagen trch t da c c mt sc chng chu c bit vi cc ph hy l v ha
hc.
Di tc dng ca cc tc nhn tia t ngoi, thuc nhum tc, thuc dui tc... tc d b tn
thng, ch ngn, mt i v bng mt. Trong cc sn phm chm sc tc thng c b sung mt lng
collagen hydrolysate c tc dng bo v cho tc, phc hi h


Hnh 3.17 Tc b h tn v phc hi sau khi s dng sn phm chm sc tc c cha collagen hydrolysate
3.4 ng dung collagen trong cng ngh nhip nh
Cc c tnh gip cho collagen c ng dng trong lnh vc ny l:
L thnh phn thit yu cho qu trnh ng t v pht trin ca dch nh tng nh :
bc halogen.
L cht n nh gia thnh phn gn kt v h nh tng thuc nhum.
L tc nhn gn kt trong qu trnh sn xut trong cc giy in phun hin nay.
Kh nng trng n tt.
C tnh bn vng cao.
CHNG 4: NHNG VN TN TI V HNG TP TRUNG NGHIN
CU GII QUYT
Trong mt s lnh vc ng dng Collagen nh trong cc ngnh cng nghip thc phm, dc
phm v m phm, i hi cht lng Collagen kh cao, khng cha tp cht, khng cha tro, khng cha
thnh phn cht bo, nht t yu cu, c trng lng phn t nht nh. Chnh v vy phi c nhng
gii php k thut x l cc vn ny.
loi cc tp cht ra khi dung dch Collagen, ta c th s dng phng php ha hc
hay kh cc ion bng mng cellophan.
tinh ch c Collagen c trng lng phn t nht nh ta dng phng php lc gel
hay tin hnh tch phn on thu Collagen.
Hin nay vn c quan tm nht trong qui trnh thu nhn Collagen l vn loi hon


ton cht bo ra khi dung dch cha Collagen. Hin nay cc nh khoa hc nghin cu thnh cng qui
trnh s dng CO2 siu ti hn trong qu trnh tinh sch Collagen.
Mt cht lng siu ti hn ( Super Critical Fluid - SCF ) l bt k mt hp cht no c gi tr
nhit v p sut trn im ti hn trong gin pha. Trn gi tr nhit ti hn th phn t kh khng
th ha lng bt k p sut no. p sut ti hn l p sut hi ca kh ti nhit ti hn. Trong mi
trng siu ti hn, ch tn ti mt pha duy nht gi l fluid. Pha fluid khng phi l pha kh cng khng
phi pha lng. N l dng trung gian gia hai pha. Pha fluid ny c kh nng ha tan tng t nh cht
lng ng thi cng c kh nng khuch tn tt nh cht kh. im ti hn l im kt thc c ng
cong cn bng pha gia hai trng thi kh v lng. Vng siu ti hn cho bit bng cch s dng s kt
hp gia s ng p v thay i nhit , gia s ng nhit v thay i p sut, hon ton c kh nng
chuyn i cc thnh phn tinh khit t trng thi lng sang trng thi kh trong vng v ngc li m
khng c s chuyn pha. Cht lng siu ti hn c kh nng ha tan nh mt cht lng v cng c kh
nng vn chuyn nh mt cht kh.
iu kin mi trng ( 25c, p sut lbar), ha tan ca mt cht trong pha kh c mi
quan h trc tip vi p sut hi ca n v nhn chung th mi quan h y khng ng k. Mt khc, trong
dung mi siu ti hn, ha tan ca mt cht tng gp 10 ln. S ha tan ca mt cht trong pha lng
siu ti hn l do s kt hp gia p sut hi v nh hng tng tc gia cht tan v dung mi. Hay ni
cch khc s ha tan ca n khng n thun ch ph thuc vo p sut.
Trong qu trnh tinh sch cht bo ra khi dung dch collagen bng cch dng CO2 siu ti
hn th lc ny, CO2 lng trong bnh cha s c dn qua thit b bm cao p nng p sut ln p sut
ti hn. Cht tr dung mi cng c dn qua thit b bm dung mi. Dng CO2 v dng dung mi c
trn ln vo nhau trc khi qua thit b gia nhit. Ti y, CO2 t c trng thi siu ti hn, dng
hn hp CO2 siu ti hn v cht tr dung mi tip tc i vo ct chit. Sau khi chit, dng cht lng siu
ti hn s mang theo cht bo i ra khi ct ti van x p sut. Khi p sut gim th lc ny cht bo tch
ra khi dng siu ti hn v dng CO2 t do s i theo ng ng ra ngoi. Ct chit lc ny ch cn li

sn phm collagen loi sch bo.
TI LIU THAM KHO
1. Fratzl, P., Collagen Structure and
Mechanics. 2008.
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Biochemistry, 78, 929-958
3. Frederic M.Richards, D.S.E., John Kuriyan, Advances in Protein Chemistry.
2005. Fibrous Proteins Coiled-Coils, Collagen and Elastomers, Volume 70
4. K. Gelse , E.Po schl, T. Aigner, 2003. Collagen - structure, function and
biosynthesis. Advanced Drug Delivery Reviews 55 1531- 1546
5. L.s. Senaratne, Pyo-Jam Park, Se-Kwon Kim, Korea, Isolation and characterisation of collagen
from brown backed toadfish (Lagocephalus gloveri) skin, 2006.
6. Shuai Zhao, M.Z., Guoying Li, Preparation of collagen from the pigskin shavings by
alkali extraction.
7. Rume Sakai, S.T., Toshizo Tsemura, Soluble collagen of chicken leg tendon; Its
dnaturation temperature and Hydrodynamic properties. 1967.
8. Nagai, T., Collagen from diamondback squid. 2003.
9. Maria Sadowska, I.K.O., Celina Niecikowska, Isolation of collagen from the skins of Baltic
cod (Gadus morhua). 2002.
10. L.s. Senaratne, P.-J.P., Se-Kwon Kim, Isolation and characterization of collagen from
brown backed toadfish (Lagocephalus gloveri) skin. 2005.
11. Food and Agricultural Organisation - Food and Nutrition Paper. 14/7/1986.
12. N.Yu. Ignat'eva, N.A.D., Determination of Hydroxyproline in Tissues and the
Evaluation of the Collagen Content of the Tissues. 2005.
13. Elzbieta Skierka, M.S., The influence of different acids and pepsin on the

extractability of collagen from the skin of Baltic cod. 2006.
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Food Chemistry, 68, 277-281.
15. Phanat Kittiphattanabawon, Soottawat Benjakul, Wonnop Visessanguan, Takashi Nagai, Munehiko
Tanaka, Thailand, Characterisation of acid - soluble collagen from skin and bone of bigeye snapper
(Priacanthus tayenus), 2005.
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Munehiko Tanaka, Characterisation of acid-soluble collagen from skin and bone of
bigeye snapper (Priacanthus tayenus). 2004.
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Hochiminh city and Cantho University, Isolation of Collagen from the skin of Basa fish (Pangasius
Bocourti).
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thermal properties ofLabeo rohitabones due to arsenic exposure. 2009.
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21. http://www.tessenderlo.com/
22. http://www.science direct.com

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