Nghiên cứu ứng dụng K-Carrageenan Oligosaccharid từ rong biển có hoạt tính sinh học trong chế biến và bảo quản thực phẩm

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Carrageenan dễ tan trong nước, các dung dịch carrageenan có độ nhớt rất lớn. Chính vì vậy, ở nồng độ rất nhỏ (1%), Carrageenan cũng có thể tạo gel. Carrageenan dễ tương tác liên kết với protein và các polysaccharid khác. Chính vì có những tính chất đặc trưng nêu trên, Carrageenan được ứng dụng trong nhiều lĩnh vực chế biến thực phẩm khác nhau.

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Nội dung Text: Nghiên cứu ứng dụng K-Carrageenan Oligosaccharid từ rong biển có hoạt tính sinh học trong chế biến và bảo quản thực phẩm

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 nhau 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 1. K- carrageenan chiet iir rong Hong Van Eucheuma geletinae. Carrageenan de tan trong nude, cac dung djch carrageenan cd do nhdt rat Idn. Chinh vi vay, d ndng do 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 trong nhieu ITnh vyc che bien thyc pham khac nhau. 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] Enzyme Phitopain do Trung tam "Ky nghe sinh hgc", Vi?n HLKH Nga cung cip 678 Tuyen tjp Hgi nghi KHOA HQC T O A N Q U Q C V £ SINH HQC BI^N VA P H A T T R I ^ N B ^ N VLTNG 2. Phuong phap nghien ciiTi - Phuang phdp xdc dinh trpng luang phdn tir m Phuang phap do 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 pho 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 pho 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 do ben cua mgt vat lieu ngudi ta su dung phucmg phap nen, ep hay keo vat do theo mgt chieu nao do tdi khi miu bj pha hiiy. D6i vdi bun, la mgt loai thyc phim nen dl xac djnh dg deo, dai ciia bun tbi hien nay 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 do 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 do xacdinh do bin nen ep. Mlu banh cd hinh fry, vdi dudng kinh: d = 6,5cm, chieu dai: 1 = 12cm. M6i mgt miu dugc cit ra 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 nay dugc xac djnh tai Trung tam ky thuat 1 thugc Tong cue tieu chuin do ludng chat lugng tren may nen ep van nang Insfron (My) vdi t i c do ep la 15mm/pbiit. - Xdc dinh cdc chi tiiu vi sinh ciia bun "^ Cac miu bim sau khi dugc cho them cac chit bao quan vdi ham lugng khac nhau dugc xac djnh cac chi tieu vi sinh tai Trung tam ky thuat 1 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 nim 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 do 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 ^ ^ ^ 9 " g lugng phan tu frung binh cua Carrageenan va Carrageenan oligosaccharid la M - 173,8 (kDa) va M = 27,7 (kDa) tuang irng. Tir ket qua thu dugc cho thiy trgng lugng phan tir tmng binh cua oligosaccharid nh6 ban so \ai trgng lugng phan tu trung binh cua Carrageenan ban diu. Dilu do chimg to thuy phan da cit ngan mach polysaccharid ciia Carrageenan. Tieu ban: HOAT C H A T SINH HOC BI^N 679 - Khdo sdt cdu true cua Carrageenan oligosaccharid bdng phuang phdp pho 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 nhau. Ket qua phan tich sy phan manh ciia chiing trong ph6 MS ciia Carrageenan oligosaccharid (binh 2) dugc the hien d bang 1. Tir dd co the m6 ta sy phan manh theo sa do fren hinh 3. . Bang 1. Su> phan manh cua Carrageenan oligosaccharid Manh Oligosaccharid m/z [Galp4S - H2O - K""]"' 241 [Galp4SAnGalp - C2H4O2 - K"]"' 341 [Galp4SAnGalp - H2O - K"]"' 385 [Galp4SAnGalp + C4H3O3 - K*]"' 467 [AnGalpGalp4SAnGalp - C2H2 - K " ] ' 503 [Galp4SAnGalpGalp4SAnGalp - C2H4O3 - K^]'' 611 [Galp4SAnGalpGalp4SAnGalp - C2H4O - K^]"' 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 2. 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 ra 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 con lai frong phan tir. Pho cpng huang tir hat nhdn. Phi cgng hudng tir hat nhan cua san phim thuy phan dugc trinh bay fren cac hinh 4, 5. 680 Tuyen tip Hoi nghj KHOA HOC T O A N Q U 6 C V £ SINH HQC BifeN VA P H A T TRifeN B £ N VCTNG M2-CARAGEENXN.OLIGOSACCHARIDE-D20-1H M2-CJUIAGEEKXH.OLIGOSACCHARIDB-D20-C13CPD Ml^ UWJ _.,, rtiiiiWi<>iiiii«wm' = f(x) va quan be giira lyc nen ngang vdi nIng dg chat phy gia z =

dung c h i t phu gia oligosaccharid Lyc nen dgc, N Lyc nen ngang, N oligosaccharid, STT A A % ya Zd y" Za 1 0,05 1425 1456 1217 1223 2 0,1 1587 1540 1267 1256 3 0,2 1661 1662 1318 1325 4 0,3 1701 1727 1402 1400 5 0,4 1745 1732 1482 1482 6 0,5 2144 1812 So Han the (0,5%) 1530 1190 Sanh 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 do cirng cua banh tinh bgt diing de lam bun. Dieu nay 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 khi nang bao quan ciia carrageenan va oligosaccharid, chiing tdi tien banh thi nghiem tren cic miu bun, dugc them cac chit phu gia khac nhau trong 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 4. Tir cac so lieu thyc nghi?m thu dugc cho thay rang chi co nhom vi khuin hieu khi phat triln manh theo thdi gian. Sau 3 ngay, t6ng s6 te bio d?it tdi 10 CFU/g frong tat ca cac mau thi nghiem. Nam men va moc bj uc che nhieu hon, sd te bao chi vugt qua 10* CFU/g. Coliforms hau nhu bj lie chi hoan toan. Nhu vay, carrageenan oligosaccharid cung nhu ban the va chitosan deu cd tic dung ire chi vi sinh vat gin tuong ty nhu nhau. 682 Tuyen tap Hpi nghi KHOA HQC T O A N Q U O C V £ SINH HQC BI^N VA P H A T T R I 6 N B £ N VCTNG Bang 4. Sy phat tri§n ciia vl sinh v^t trong thyc phim khi bao quan 1 Oligosaccharid Carrageenan Han the Chitosan sau 24 gio' Tong vi khuan hiiu khi, CFU/g 8,5.10^ 8,6.10^ 6,7.10^ 2,6.10^ Tong n i m men + moc, CFU/g 5,1.10' 4,6.10" 3,8.10" 2,3.10" Tong Coliforms, MPN/g Khong phat hien Khong Khong Khong sau 48 gid Tong vi khuan hieu khi, CFU/g 1,3.10' 8,8.10' 1,3.10' 5,9.10^ Tong nam men + moc, CFU/g 2,9.10^ 1,7.10^ 2,6.10^ 2,8.10^ Tong Coliforms, MPN/g Khong phat hien 0,36 Khong 0,36 sau 72 gio Tong vi khuan hieu khi, CFU/g 1,5.10' 9.10' 1,4.10' 9.10' Tong nam men + moc, CFU/g 2,4.10^ 2,7.10^ 2,6.10^ 2,6.10^ Tong Coliforms, MPN/g Khong phat hi^n 0,36 0,36 0,36 TV. KET LUAN Tir cac ket qui nghien ciiu, chiing t6i rut ra 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 khi 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 do ciing ciia banh tinh bgt diing dl lam biin. Ham lugng t6i uu oligosaccharid nay dl tang do ciing cua tinh bgt cd gia tri la 0,2%, frong khi 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 trong sin xuit cac sin phim lam tir tinh bgt (bun). V. TAI LIEU THAM KHAO 1. 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) .ic-Carrageenan - Protein hiteractions: Effect of Proteins on Polysaccharide Gelling and Texfriral Properties. Lebensmittel- Wissenschaft und-Techiwlome Volume 35, Number 8, pp. 741-747(7) 5. Tijssen R L M, L S Canabady-Rochelle, M Mellema (Jun 2007). Gelation upon long storage of milk drinks with carrageenan. J Dairy Sci. (Vol. 90, Issue 6, Pages 2604-11). 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 1 / 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: Xic 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. STUDY ON T H E USE O F T H E BIOACTIVE 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 In this study, we demonstrate that k-carrageenan oligosaccharides were prepared by enzymatic digestion of kappa-carrageenan from red seaweeds Hong Van Euchema gelatinae. And recently il is shown that Carrageenans and carrageenan oligosaccharides are an effectively additional substrate for the processing and preservation of food derived from rice starch.