Trgng lugng phan tir frung binh cua Carrageenan va oligosaccharid dugc xac dinh theo phuang phap do dg nhdt bing nhdt kl Ubeldt.. Tir ket qua thu dugc cho thiy trgng lugng phan tir tmng[r]
(1)Tieu ban: HOAT C H A T SINH HQC BI^N 677
NGHIEN ClTU tTNG DUNG K - C A R R A G E E N A N
OLIGOSACCHARID TlT RONG BIEN CO HOAT TINH SINH HOC TRONG CHE BIEN VA BAO QUAN THl/C PHAJM
Tran Dinh Toai'*, Pham Hong Hai', Nguyen Van Thiet^ ' Viin Hod hpc, Viin Khoa hpc vd Cong nghi Viet Nam
^Vien Cong nghi Sinh hpc, Viin Khoa hpc vd Cong ngh? Viit Nam I M d DAU
Carrageenan (Car) la polysaccharide ciia galactose ggi la galactan Galactose frong mach galactan cd thi la a (l-3)-D-galactose (G), |3(I-4)-D-galactose (D) hoac P(l-4)-3,6-anhydro-D-galactose (DA)[1]
Carrageenan dong thdi la sulfat polysaccharide Ngoai mach polysaccharide chinh edn cd the CO cac nhom sulfat dugc gan vao d nhung vj tri va s6 lugng khac tao nen cac sulfat polysaccharide va co ky hieu rieng nhu: X - , K - , i - , v - carrageenan [2]
Vi du, carrageenan tir rong H6ng Van Euc gelatinae dugc xac lap la K- carrageenan (hinh 1)
[3]-Hinh K- carrageenan chiet iir rong Hong Van Eucheuma geletinae
Carrageenan de tan nude, cac dung djch carrageenan cd nhdt rat Idn Chinh vi vay, d ndng rit nhd (1%), carrageenan ciing cd the tao gel Carrageenan de tuong tac lien kit vdi protein va cac polysaccharid khac [4] Chinh vi co nhimg tinh chat dac trung neu fren, Carrageenan dugc img dung nhieu ITnh vyc che bien thyc pham khac Dac biet, carrageenan khdng the thieu dugc frong c6ng nghiep che bien sua [5] Carrageenan Oligosaccharide cd thi thu dugc bang each dimg enzyme thuy phan Carrageenan [6] Carrageenan cung nhu oligosaccharid ciia no cd boat tinh sinh hgc rat cao [7]
Do do, chung toi thiiy phan Carrageenan tao oligosaccharide va nghien ciiu img dung lam phu gia bao quan, chi bien thyc pham (biin tir tinh bgt gao)
H NGUYEN LIEU VA PHU'ONG PHAP NGHIEN c i r u I.Nguyen lieu
K- carrageenan chiet tir rong H6ng Van Eucheuma geletinae [3]
(2)2 Phuong phap nghien ciiTi
- Phuang phdp xdc dinh trpng luang phdn tir m
Phuang phap do nhdt de xac djnh trgng lugng phan tii trung binh cua Carrageenan va Carrageenan oligosaccharid theo he thiirc Mark - Houwink: [T]] = K.M"
- Phuang phdp xdc lap cdu true
Cac phuang phap ph6 cho phep xac lap cau friic cua carrageenan va oligosaccharid
Phuang phap ESI - MS (phuang phap sic ki long khii co bg kit ndi phun dien)
(Negative-ion electrospray ionisation-mass spectrometry) [9]
Pho cgng hudng tir hat nhan [10]
- Khdo sdt dp bin nen ep ciia banh tinh bpt
Ve nguyen tac, de xac djnh ben cua mgt vat lieu ngudi ta su dung phucmg phap nen, ep hay keo vat theo mgt chieu nao tdi miu bj pha hiiy
D6i vdi bun, la mgt loai thyc phim nen dl xac djnh dg deo, dai ciia bun tbi hien chua CO may nao thyc hien chiic nang nay, fren thyc tl ngudi ta danh gia dg deo, dai ciia bun bing lay phieu tham cua nhurng ngudi tieu dimg san pham Do do, chiing tdi chi tao miu banh tinh bgt CO hinh fru lam vat lieu va tir xacdinh bin nen ep Mlu banh cd hinh fry, vdi dudng kinh: d = 6,5cm, chieu dai: = 12cm M6i mgt miu dugc cit lam hai mau nhd cd chieu dai: 1, = 6cm Mgt mau dugc nen theo chieu dgc, mgt miu dugc nen theo chilu ngang
Do ben nen ep dugc xac djnh tai Trung tam ky thuat thugc Tong cue tieu chuin ludng chat lugng tren may nen ep van nang Insfron (My) vdi t i c ep la 15mm/pbiit
- Xdc dinh cdc chi tiiu vi sinh ciia bun
xac
"^ Cac miu bim sau dugc cho them cac chit bao quan vdi ham lugng khac dugc
djnh cac chi tieu vi sinh tai Trung tam ky thuat thugc Ting eye Tieu chuin Do ludng Chit lugng
Cac chi tieu vi smh bao gIm: ting sd vi khuin hilu khi, tong nam men va n i m moc tone
Coliforms [U] ' ^ - Phuang phdp xir ly s6 liiu
Ket qua thyc nghi?m dugc xu ly theo phuang phap binh phuang toi thilu tren may tinh dl tim phuang trinh h6i quy m6 ta tuang hgp kit qua nhan dugc Phan tich thong ke dugc thuc hi?n theo cac tieu chuan Student va Fisher [12]
III KET QUA NGHIEN ClTU
1 Nghien cihi thu nhan oligosaccharid
De thu nhan Carrageenan oligosaccharid, tiln hanh thiiy phan Carrageenan bing dung dich enzyme a nhiet dg 60 C Trgng lugng phan tir frung binh cua Carrageenan va oligosaccharid dugc xac dinh theo phuang phap dg nhdt bing nhdt kl Ubeldt
Vdi cac gia trj K va a dac frung da bilt, theo he thiic Mark - Houwink: [ri] = K M" xac ! ' " ' ; , ^ J ^ ^ ^ " g lugng phan tu frung binh cua Carrageenan va Carrageenan oligosaccharid la M - 173,8 (kDa) va M = 27,7 (kDa) tuang irng
(3)Tieu ban: HOAT C H A T SINH HOC BI^N 679
- Khdo sdt cdu true cua Carrageenan oligosaccharid bdng phuang phdp
Pho khoi luang
Do Carrageenan ciing nhu oligosaccharid deu la polysaccharid da dien tich nen ph6 kh6i ciia chiing rat phirc tap bao g6m nhieu dinh va cac dinh lai chen sat Ket qua phan tich sy phan manh ciia chiing ph6 MS ciia Carrageenan oligosaccharid (binh 2) dugc the hien d bang Tir dd co the m6 ta sy phan manh theo sa fren hinh
Bang Su> phan manh cua Carrageenan oligosaccharid
Manh Oligosaccharid [Galp4S - H2O - K""]"'
[Galp4SAnGalp - C2H4O2 - K"]"' [Galp4SAnGalp - H2O - K"]"' [Galp4SAnGalp + C4H3O3 - K*]"' [AnGalpGalp4SAnGalp - C2H2 - K " ] '
[Galp4SAnGalpGalp4SAnGalp - C2H4O3 - K^]'' [Galp4SAnGalpGalp4SAnGalp - C2H4O - K^]"'
m/z 241 341 385 467 503 611 647
Display Repx>rt - Selected Window Selected Analysis
lAo 3ba xo SJL AA al.iU.iS>»7rv ••
M50 T r ^ i (teport V « (At 0|K2)
Hinh Pho MS cua Carrageenan oligosaccharid
Tir kit qua tren ta thay nhung manh c6 tin suit cao fren ph6, the hifn rang viec phan manh ciia chung tuin theo ca che phin manh ciia cacbohydrat da dugc c6ng b6 frudc day [13] Sy phan minh cua chiing chii yeu xay d lien ket glycosit giira cac monomer Day la nhirng vi tri ma nang lugng lien ket ciia chiing thap ban so vdi nhimg lien ket lai frong phan tir
Pho cpng huang tir hat nhdn
(4)M2-CARAGEENXN.OLIGOSACCHARIDE-D20-1H M2-CJUIAGEEKXH.OLIGOSACCHARIDB-D20-C13CPD
Ml^
UWJ _.,,
%
Hinh Ph6 1H- NMR cua Carrageenan '^ Oligosaccharid
rtiiiiWi<>iiiii«wm<imni
Hinh Pho 13C-NMR cua Carrageenan Oligosaccharid
Phan tich cac tin hieu phi NMR, xac djnh chuyen djch hda hgc cua cac mau Oligosaccharid nhu sau:
Bang 2: Do chuyen dich hda hgc 13C NMR va 1H NMR ciia m i u Oligosaccharid
P(1,3)-D-galactose
p(1,4)-3,6 anhydro galactose
C1 H1 103,03 5,043d 95,72 5,527d
C2 H2 70,13 4,031i 70,43 4,625br
C3 H3 79,34 4,397dd 79,73 4,942d
C4 H4 74,46 5,255d 78,87 5,043d
C5 H5 75,31 4,228br 77,31 5,063s
C6 H6 61,82 4,228br 70.01 4,560br
Qua vifc phan tich va so sanh cac dir lieu ve ph6 cgng hudng tir hat nhan cua Carrageenan [3] va Oligosaccharid cho thay qua trinh thuy phan khong lam thay d6i don vj ciu tnic ma chi pha vd cac lien ket glycosit lam cat ngan mach polymer ciia Carrageenan
2 Nghien cuu sir dung Carrageenan oligosaccharid lam phu gia bao quan, che bien thuc pham (bun lam tir tinh bot gao)
- Khdo sdt tdc dung ciia Carrageenan oligosaccharid lam tdng dp ciing cua banh tinh bpt Tac dung ciia carrageenan oligosaccharid lam tang cung cua banh tinh bgt dugc the hi?n d ben nen dgc, nen ngang cua banh tinh bgt
Cac ket qua xac djnh lyc nen dgc va lyc nen ngang tac dgng tdi miu banh tinh bdt dl danh gia tac dung ciia phu gia oligosaccharid dugc ghi frong bang
Ket qua tien hanh xu ly s6 lieu tim phuang trinh hoi quy md ta quan hg giira lyc nen dgc vdi nong dg chat phu gia>' = f(x) va quan be giira lyc nen ngang vdi nIng dg chat phy gia z =
<p (x) cho thiy cac quan he cd dang phuang trinh tdng quat cua mgt parabol:
A
Cac he s6 bo, b|, b2 dugc xac djnh theo phuong phap binh phuong toi thilu
(5)Tieu ban: HOAT C H A T SINH HOC BI^N 681
0 CO gia tri tir den va cang Idn thi quan he phi tuyen tim dugc cang co tin ciy cao Cac sd lieu bang 3dugc xir ly tren may tinh theo phuang phap da neu cho kit qua nhu sau:
P h u a n g trinh boi quy m6 ta sy phu thugc vao ham lugng oligosaccharid ciia lyc nen dgc va lyc nen ngang deu co dang phi tuyen nhu sau:
- P h u a n g trinh boi quy d6i vdi lyc nen dgc:
ya = 1358,7-I-2107x2 - 2932,1x2^ Vdi: = 0,967 - Phuang trinh hdi quy ddi vdi lyc nen ngang:
Zd= 1192,2-1-604x2-^-299,5x2^ Vdi: = 0,99 Cac sd lieu thyc nghiem va tinh toan ciing dugc dua vao bang
Bang K i t qua thyc nghifm va tinh toan cac thdng sd dg cii'ng cua banh tinh bpt su> dung c h i t phu gia oligosaccharid
STT
1
So Sanh
oligosaccharid,
%
0,05
0,1 0,2 0,3 0,4 0,5
Han the (0,5%)
Lyc nen dgc, N
ya
1425 1587 1661 1701 1745 2144 1530
A
y"
1456 1540 1662 1727 1732
Lyc nen ngang, N
Zd 1217 1267 1318 1402 1482 1812 1190
A
Za
1223 1256 1325 1400 1482
Qua nhimg kit qua thyc nghiem thu dugc cho thay rang oligosaccharid thyc sy la chit phy gia chi biln va bao quin co tic dyng lam tang cirng cua banh tinh bgt diing de lam bun Dieu co thi giii thich nhu sau: oligosaccharid cd mach ngan, de tan frong nude nen de dang phan tan lam tang sy tuong tie vdi tinh bgt tao cho tinh bgt co dg cung cao ban, ben chac hon
- Cdc chi tieu vi sinh
D I danh gia nang bao quan ciia carrageenan va oligosaccharid, chiing tdi tien banh thi nghiem tren cic miu bun, dugc them cac chit phu gia khac qua trinh sin xuit Ket qua so sinh hieu qua Ore che sy phat trien ciia vi sinh vat cua cac chit phy gia dugc ghi frong bing
(6)Bang Sy phat tri§n ciia vl sinh v^t thyc phim bao quan
1 Oligosaccharid Carrageenan Han the Chitosan
sau 24 gio' Tong vi khuan h i i u khi, CFU/g
Tong n i m men + moc, CFU/g Tong Coliforms, MPN/g
8,5.10^ 5,1.10' Khong phat hien
8,6.10^ 4,6.10" Khong
6,7.10^ 3,8.10" Khong
2,6.10^ 2,3.10" Khong sau 48 g i d
Tong vi khuan hieu khi, CFU/g Tong nam men + moc, CFU/g Tong Coliforms, MPN/g
1,3.10' 2,9.10^ Khong phat hien
8,8.10' 1,7.10^ 0,36
1,3.10' 2,6.10^ Khong
5,9.10^ 2,8.10^ 0,36 sau 72 gio
Tong vi khuan hieu khi, CFU/g Tong nam men + moc, CFU/g Tong Coliforms, MPN/g
1,5.10' 2,4.10^ Khong phat hi^n
9.10' 2,7.10^
0,36
1,4.10' 2,6.10^ 0,36
9.10' 2,6.10^
0,36
TV KET LUAN
Tir cac ket qui nghien ciiu, chiing t6i rut cic ket luan sau:
1 Thiiy phan kh6ng lam thay d6i ciu true, chi cd tic dung phi vd cac lien kit glycosit lam cat ngan mach cua Carrageenan Sau thuy phin, Carrageenan oligosaccharid thu dugc co kh6i lugng phin tir tmng binh (27,7 kDa) giim so vdi khii lugng phin tii trung binh ciia Carrageenan ban diu (173,8 kDa)
2 Carrageenan oligosaccharid co tie dung lam tang ciing ciia banh tinh bgt diing d l lam biin Ham lugng t6i uu oligosaccharid dl tang ciing cua tinh bgt cd gia tri la 0,2%, frong ham lugng t6i uu cua phu gia Carrageenan cung nhu bin the la 0,5%
3 Nhu vay, Carrageenan oligosaccharide co t h i diing lam phu gia che biln va b i o quan thay the co hieu qui va kh6ng dgc hai so vdi b i n the sin xuit cac sin phim lam tir tinh bgt (bun)
V TAI LIEU THAM KHAO
5
Thanh Thj Thu Thuy (2002) Stmcture of sulfated polysaccharides in aqueous solufron Doctoral thesis submitted to Kyoto Institute of Technology, Japan
Christian W Kasbauer, Dietrich H Paper, Gerhard Franz (2001) Sulfated y0-(l-^4)-galacto-ohgosachharides and thefr effect on angiogenesis Carbohydrate Research, 330 427-430
Trin Dinh Toai, Nguyin Xuan Nguyeri, Pham Hong Hii, Nguyen Bich Thuy, Trin Thj H6ng (2006) Carrageenan tir rong biln: san xuit va ling dung Nxb KH & KT Ha Ngi^
100 frang
Baeza R I.; Carp D J.; Porez O E.; Pilosof A M R (December 2002) icCarrageenan -Protein hiteractions: Effect of -Proteins on Polysaccharide Gelling and Texfriral Properties
Lebensmittel- Wissenschaft und-Techiwlome Volume 35, Number 8, pp 741-747(7)
(7)Tieu ban: HOAT C H A T SINH HQC BigN 683
6 Haijin M., Xiaolu J.; Huashi G A K-carrageenan derived oligosaccharide prepared by enzymatic degradation containing anti-tumor activity, Joumal of Applied Phycology Volume 15, Number 4, July 2003, pp 297-303(7)
Huamao Yuan and Jinming Song Preparation, structural characterization and in vitro antitumor activity of ^ap/?a-carrageenan oligosaccharide fraction from Kappaphycus
striatum Joumal of Applied Phycology Volume 17 Number / January 2005 pp 7-13
8 Vu Nggc Ban Giao trinh thyc tap Hoa ly (1989) DH KHTN-Ha Ngi
9 Ekeberg D, Knutsen S H, Sletmoen M (Aug 2001) Negative-ion elecfrospray ionisafron-mass specfrometry (ESI-MS) as a tool for analysing stmctural heterogeneity in kappa-carrageenan oligosaccharides Carbohydr Res (Vol 334, Issue 1, Pages 49-59)
10 Gutierrez A M., Kumosinski T F Farrell H M., Jr (1998) K- Carrageenan interaction with bovine and caprine caseins as shown by sedimentation and nuclear magnetic resonance specfroscopic techniques J Agric Food Chem., 46, 4987-4996
11 Cac Tieu chuan Viet Nam: TCVN 5165-90: X i c djnh tong s i vi khuin hilu khi; TCVN 7137: 2002 Xac djnh ting nam men va nim mic; TCVN 7135:2002 Xac djnh tong
Coliforms
12 AxnaaapoBa C Jl., Kaijiapoa B B OnTHMHsauHa 3KcnepHMeHTa B XHMHH H xHMHMecKOH TexHO.nornH, 1978 Hsfl "Bbicmaa mKona", MocKsa
13 Yu G, Guan H loanoviciu AS Sikkander SA Thanawiroon C Tobacman JK Toida T Linhardt RJ (2002) Structural studies on kappa-carrageenan derived oligosaccharides
Carbohydr Res; 331:433 AO
S T U D Y O N T H E U S E O F T H E B I O A C T I V E K - C A R R G E E N A N
O L I G O S A C C H A R I D F R O M R E D S E A W E E D S AS AND A D D I T I O N A L S U B S T R A T E F O R T H E F O O D P R O C E S S I N G AND P R E S E V A T I O N
Tran Dinh Toai, Pham Hong Hai, Nguyen Van Thiet
SUMMARY
Carrageenans, a common cell wall polysaccharide, are a family of linear sulphated
polysaccharides exfracted fi-om red seaweeds Carrageenans are large, highly flexible molecules
which curl forming helical structures This gives them the ability to form a variety of different
gels at room temperature They are widely used in the food and other industries as thickening
and stabilizing asents Carrageenan oligosaccharides has this high effect thanks to thefr smaller molecule weight as compared to the one of Carrageenan