THONG BAO KHOA HOC
DANH GIA HIEU QUA SU" DUNG ENZYME VISCOZYME L THAY THE HOA CHAT TRONG SAN XUAT CARRAGEENAN
Tir RONG SUN KAPPAPHYCVSALVAREZII (DOTY) DOTY
EVALUATING THE EFFECT OF USING VISCOZYME L ENZYME REPLA CING CHEMICALS IN PRODUCTION OF CARRA GEENAN
FROM SEAWEED KAPPAPHYCUS ALVAREZH (DOTY) DOTY
Bui Huy Chich', Bd Van Ninh', VS Ngpc Bg?
Ngfty nhiin bai: 16/4/2014; Ngay phan bi?n thong qua. 16/10/2014. NgSyduyel dang: 01/12/2014
T O M T A T
Phircmgphdp sir dung enzyme Viscozyme L xU ly rong trang qud trinh sdn xudt carrageenan tu rong Kappapkyaa alvarrezii (Doty) Doty co nhieu uu diem so vai phuang phdp xu ly rong bdng NaOH. Cu the, su dyng enzyme Viscozyme L xu ly rong cho hieu sudt quy trinh vd sue dong carrageenan cao han, khac phi^ic nhung han che cua qud trinh sdn xudi carrageenan theo phuang phdp sii dung NaOHxu ty rong nhu carrageenan thu duac thuong Idn hoa chdt. qud trinh tinh che rat kho khan. 6 nhiem moi tru&ng, dgc hai cho ngu&i sdn xudt
Tukhoa- Kappaphycus alvarezii (Doty) Doty, carrageenan. Viscozyme L ABSTRACT
Method of using Viscozyme L enzyme for treating seaweed in the production of carrageenan from seaweed Kappaphycus alvarrezii (Doty) Doty has several advantages over the NaOH treatment method. Specifically. Viscozyme L enzyme treatment of seaweed gave higher yield and higher carrageenan gelation degree, overcoming the disadvantages of carrageenan production by NaOH treatment where carrageenan obtained was often mixed with chemicals, difficuU ta be refined, causing environmental pollution, and harmful to workers
Keywords. Kappaphycus alvarezii (Doty) Doty, carrageenan. Viscozyme L
I. DAT V A N O £ Hi$n tgi, cdc nghien cuu tap trung vdo ITnh vyc Rong sun Id nguyen lieu de sdn xual nuoi trong vd thu nhgn Carrageenan tir rong syn Iheo Carrageenan, mpt loai polysaccharide duoc b i l l vd quy trinh xii 1^ rong bang acid hay kiem. Phuong su dung tu lau, ij-ng dyng trong rai nhieu ITnh vuc phdp xt> ly ndy sdn xual Carrageenan c6n co mpt nhu ch4 biln sua, thjt, my pham, lhu6c.. Tuy nhien, so hgn c h i Id Carrageenan thu duac thudxng lin vdi c5ng nghe che bien rong syn vd Carrageenan h6a chat ndn qua trinh tinh che ggp nhilu kho khdn, to rong stjn a Vi§t Nam con han che nen rong stjn 6 nhiem m6i trud'ng, dpc hgi v6'i nguc^i sdn xuat.
nguyen lieu it fluo'c tieu thy Irong nud-c md chO Do vdy, d6i hoi birc xtic cua thuc tien ddt ra Id can yeu chij y l u xuat khau qua thuong ldi dual dgng nghien ct>u phat trien de co dugc cong nghd sar nguyen lidu kho ngn rat d l bj ep gid, bi dong tr6ng xult Carrageenan tu rong syn hi^u qua, on dinti, cho' vdo th! truang nuac ngodi Trong khi, hdng nam hgn che vi§c si> dyng hoa chat Idm anh huong der dat nudc ta van phdi ton mOt lup'ng ngoai te Ion d l an loan thuc pham vd 5 nhiem moi trudng cung c^i:
nhap kh^u cac loai Can-ageenan Idm phtj gia cung cho cdc ngdnh cong nghi?p trong nuoc vd hudng cdp cho cdc ngdnh c6ng nghiep trong nud'c [1] d i n xuat khau, hgn c h i dupc tinh trang xu^t khai
1 Th S BCii Huy Chich; Truing Cao dang cpng Song BS R|a Vflng Tdu
^ TS. D6 van Ninh: Trufyng O^i hgc Nha Trang
^ TS. Vu Ngoc Bpi: Khoa C6ng nghe thyc phim - Truing Ogi hpc Nha Trang
10 • TRUONG DAI HOC NHA TRANG
Tap chi Khoa hoc - Cong nghe Thuy sdn So 4/2014
rong nguyen lidu kh6 vdi gid re, nhdp khauCan^geenan v6'i gid cao nhu hi#n nay. Ti> do, gop ph^n gidi quylt dau ra cho nghl nuoi trong rong sun, ndng cao gid trj thuong pham. Thtjc day nghl trong rong syn tgi Vift Nam phdt trien.
Vol phuong phap su dyng enzyme d l xu ly rong, phdn i>ng xdy ra trong dilu ki#n nhe nhdng, hieu qua cao do tinh dgc hi#u cua enzyme [6], khong dgc hai cho nguai san xuat va dnh hudng den moi trudng, ddm bao an loan thyc phUm dang Id mOt hudng nghien cuu duac nhilu nhd khoa hpc trong vd ngodi nudc quan tam.Trong phgm vi bdi bdo nay, chung 161 khong trinh bdy qud trinh nghien cuu d l tu do de xudt quy trinh sdn xuat carrageenan ICF rong syn theo phuo'ng phap xu ly rong bang enzyme Viscozyme L ma su dyng luon quy trinh da nghldn ci>u d l ddnh gia hi^u qua su dung enzyme Viscozyme L thay t h i hoa chdt tnang sdn xuat carrageenan ttj nang sun.
II. D 6 | TlfONG VA PHUONG P H A P NGHIEN CLTU 1. Doi tu'ong nghien CLPU
- Nguyen lieu: Rong syn Kappaphycus alvarrezii (Doty) Doty dugc nuoi trong tgi d^m Thuy Trieu, xa Cam DiJc, huyen Cam Lam, tinh Khdnh Hoa. Thdi gian thu mau, ngay 4/4/2013, rong co dp tuoi la 45 ngay. Sau dd, rong dup'c chuyin v l phdng thi nghiem Trudng Dai hgc Nha Trang phoi khQ tdi dO I m ^ 25%, vd duwc su dyng de lien hdnh tdch chilt Can-ageenan.
- Enzyme Viscozyme L: Ctia hang Novozymes do Cong ty Nam Giang -133/7 H i Vdn Hud, Phudiig 9,
PhO Nhulln, TP. HCIVt cung d p , hogt dp 100 FBG/g, dilu kidn thfch hop d l c h i pham hogt dOng:
pH 3,5 - 5,5 vd nhlgl dp 40 - SS^C. Bdo qudn c h i phim dnhlgt d p i -lO^C.
2. Phuo'ng phdp nghien ci>u 2.1. Ciic phuong phap phan tich
- Xdc djnh hdm I m : bang phuang phdp sly den khli lupng Wiong doi theo tieu chuan TCVN 3700 - 90.
- Xdc djnh SLPC dong theo Craigle (1978) [5].
- Phuang phdp nhupm kdp: Dung d l phdn bl^t mgt cdeh ro rang cac mo cua mOl ca quan. NhCfng mo c6 mdng le bao hoa go se bdt mau xanh cCia pham lye lode hogc xanh methylene. Nhung m5 c6 mdng t l bdo c l u tgo bang cellullose, se bat mdu hing cua phim d6 carmin [4].
- Xdc djnh hi^u suit thu hoi Carrageenan theo cdng thi>c [3]:
A(100-Wj)
^ = P(ioo-w,) ^oo"/"
Trong do, A: Id so gram Carrageenan thu dugc (g);
P: So gram rong dem nlu chilt (g), W,: Op am cua rong nguyen lieu (g); W^; Do I m cua Carrageenan (g).
2.2. Phwcmg phap bo tri thi nghi$m
Tien hanh thi nghiem sdn xult can^geenan theo hai quy trinh' Xil ly rong bdng enzyme Wscozyme L va san xuat carrageenan theo phuang phap xu ly rong bang NaOH tren ctjng mdu rong nghien cCru. Vdi mol quy trinh sdn xuat, ddnh gid hi§u suit thu hll san pham, sue dong, pH vd mdu sac san phim thu dug-c.
Rong syn kho
- Ty le nudc nlu/rong 50/1 - t"c= ds^C:
- Thdi gian nau 70 phtJt
I}^
L
Ngam (1 - 2h)
Lpc ndng Bo sung KCI
IE
Cap dong Ra ddng Phai hogc sly
Carrageenan D i d d n g , cat spi
:
- Ty le rong/nudc: 20/1 - Nong dg enzyme: 1%
- Nhipt do xu ly 45^0 - pH = 4,6
Hinh 1. San xuit carrageenan theo phirong ph^p xv 1^ rong bang enzyme Viscozyme L (Bui Huy Ctiich, Do Van Ninh vk cs, 2013)
TRUONG OAI HOC NHA TRANG • 11
Tgp chi Khoa hpc - Cong ngh? Thuy sdn SdJ/m
Giai thich hinh 1: Rong syn kh6 dupc n>a sgch, ngdm truong nd ttj 1 - 2 gid, tilp d6 dem xt> 1;^ bang Viscozyme L vdi cdc thSng so: Lupng nudc g i p 20 lan trgng lupng rong kho; Ty Ig enzyme/rong:
1%; Nhigl dp' 45''C; pH moi tnjdng: 4,6; Thdi gian:
60 phut. Sau khi xu ly enzyme rong dupc dem di riJa
nhdm logi bo tgp chat tao ra trong qua trinh thuy pha^
enzyme. Tilp theo, rang dupc dem di nlu chldt 6 95»C, ty Ig nudc/rang kho; 50/1 vd thdi gian 70 phOt Hon hpp dupc Ipc qua mgt Idp vdi, r6i b l sung 0,3%
KCI vap d|ch Ipc. d l ddng tu nhifen, cdt milng, ^ ddng, ra dong vd Idm khO thu sdn pham carrageenan,
-Nhiet dp 102''C - Thdi gian 65 phiit - Ty le nudc/rong 52/1
1 Rong syn kh6
i
Ngdm, n>a
1
1 1 1
1 Xi> \'j/ kiem r ~ \
Rua trung tinh
i I
] i
^ — » | Nau chiet
1 i
LocDjch loc
i i
Ldm dong, tan gia Phai/ say
i
1 Xay nghien
i 1
Bao got
n J
~^^
- Nhi^t d$ 30''C -Thdi gian 40 phut - Nong dd NaOH 6%
1 . 1 - - 1
__M-
DK
n 1 1 1
KCI 0.06% 1
Hinh 2. San xuat carrageenan theo phirmig phdp m ly rong bing NaOH (Dio TrQug HUU, 2007) Giai thich hinh 2. Rong syn kho dupc rua sgch,
ngdm truang nd tu 1 - 2 gid, tiep dd dem XCF ly vdi NaOH vdi cdc thdng so: Lupng nudc gap 20 lan trang luang rong khd, Nong dg NaOH: 6%; Nhigl Q6- SO'^C; Thdi gian. 40 phut Sau khi xu ly NaOH rong duac dem di lira Irung Knh bdng HCI nham logi bd tap chat va trung hda kilm du. Tilp theo, rong duac dem dl nlu chilt a 102"C. ty le nudc/rong kho:
52/1 vd thdi gian 65 phut. Hdn hpp dupc Igc qua mOt Idp vai, roi bo sung 0,06% KCI vao dich Igc, de ddng tu nhidn, c l t milng, cap dong, rd dong vd Idm kh5 thu sdn pham carrageenan [2].
Song song, tiln hdnh chup hinh dnh vd Idm tieu bdn doi vdi rong trudc, sau khi xtj ly NaOH, enzyme Viscozyme L de quan sat su thay doi cua thdnh t l bdo rong.
Deed dupc hinh anh cua rong sun trudc va sau
khi xu ly vdi kiem, enzyme Viscozyme L su dyng phan mem mdy anh cua dign thoai iphone 4s.
D I quan sdt st/ thay doi ctJa thdnh te bao rong trudc, sau khi xu ly NaOH, Viscozyme L diing phuang phdp nhugm kep vd quan sdt dudi kini' hiln VI di§n tu vdi do phdng dai 100X.
III. KeTQUANGHl6NCI>U
1. Ket qua quan sdt hinh anh thdnh te bao roni trifijc va sau khi xir ly
Hinh dnh cCia rong syn trudc vd sau khi xi> I' vdi kiem, enzyme Viscozyme L t h i hi#n d hinh 3 hinh 4 vd hinh 5.
Vi dnh thdnh l l bdo rong trudc va sau khi xi ly NaOH vd Viscozyme L quan sdt dudi kinh h i i vl bdng phuang phap nhupm kep dupc t h i hl$n i hinh 6, hinh 7 vd hinh 8.
12 • TRUONG DAI HOC NHA TRANG
Tap chi Khoa hoc - Cong nghe Thiiy sdn So 4/2014
Hinh 3. Anh chyp rong sun khi chu^ xijr K
- • v * . i ^
Hinh 4. Anh chyp rong syn sau khi xir 1;^
NaOH 6%, V thv^g, x^ 40 phUt
Hinh 5. Anh chup rong sun khi %v ly bang enzyme I %, T ^ 45 "C, pH= 4,6. T= 60 phiil 1.1. Mau sBc than rong
Mdu sdc cua thdn rong xt> ly bdng phuang phdp NaOH cd mau trang sang han (hinh 3, 4, 5) Do khi dtJng kilm (hinh 4) cd tdc dung bdo mdn thdn rong, pha vd nhieu Idp t l bao sac to, khu sac to trong rong nen than rong thu dupc cd mdu trdng sdng.
Trong khi dd, diJng enzyme (hinh 5) thi khi bdo m6n than rang Ihi mgt phan nd da phd vd Idp t l bdo sdc to nhung v i n chua tridt nen sac to v i n cdn luu lai mOt it trong than rang nen mdu sdc t l i han.
1.2. Kich thuoc than rong
Sau khi xu ly, kich thudc thdn rong c6 sv thay do! ro rgl, cd the nhgn bilt bdng mdt thudng (hinh 3, 4, 5). Cy t h i , kich thudc thdn rong sau khi xu 1;^
Hinh 7. Vi inh rong sun san klii xir ly bang NaOH 6%, T° thirimg, T= 40 phiit
Hinh 8. V i anh rang sun klii xir I j b i n g enzyme 1 % , 1*= 4S°C, pH = 4,6, T = 60 phiit vdi NaOH 6% gidm nhieu nhdl sau dd d i n than rong xu 1;^ bang enzyme. Do khi xi> ly rong bang NaOH vd enzyme ed tdc dyng bao mdn than rong Tuy nhidn, ngodi tdc dyng bao mdn than rong NaOH cdn tan cdng vdo cdc Idp t l bdo ben trong. Ddy cung Id ca sd d l glal thich nguyen nhan hieu suat thu hoi carrageenan eua phuang phdp xi> ly bdng NaOH thip han phuang phap xu ly bang enzyme.
1.3. Kich thuoc thanh te bao cua rong sun Thanh te bao ciJa rong sau khi xu ly deu gidm ro rdt, luy thudc vdo phuang phdp xi> 1;^ rong khac nhau ma muc dp gidm khde nhau. Doi vdi phuang phdp xO iii rong bang NaOH (hinh 4, 7) thi thanh t l bdo mat hdn vd cd t h i NaOH t i n cdng cdt
TRUC7NGDAI HOC NHA TRANG • 13
Tfip chi Khoa hoc - Cong nghe Thuy sdn
mgch Carrageenan. D 6 I vdi phuang phdp xu 1^ rong bang enzyme Viscozyme (hinh 5, 8) thanh le bao giam so vdi khi chua xu ly, c6 t h i mIt hin.
2. Ket qud danh gia c h i t luwng Carrageenan theo phuxyng phdp xi> ly bang enzyme vo-i p h y c n g phap
XLF 19 bang hda c h i t , Kit qud ddnh gid chit lupng Carrageenan theo phuang phdp xu 1;^ bing enzyme vdi phuang phdp xu^
bang hoa chit dupc t h i hi§n d bdng 1.
Kit qud d bdng 1 cho thiy:
Bang 1. Ket qud ddnh gia c h i t luwng Carrageenan theo phuvmg phap xi> IJ bdng enzyme vd-l phu'O'ng phdp x u ly bdng hda c h i t
STT
1
2
Mlu
Xi> ly bing NaOH
XLT I^ bang enzyme Viscozyme Hi#u sudt
16,76
29,31 Si>c d6ng
680±5
617±5 pH (Carrageenan 1%)
8,6
6,2
Mdu s i c
S tts^
2.1. V4 m$t hieu su^t
Sir dyng c h i phim enzyme Viscozyme L thu dupe Carrageenan eo higu suit thu nhdn cao han khi dung hoa chit Do enzyme Viscozyme L eo tdc dyng bao mdn thdnh l l bdo cua thdn rong, giOp cho qud trinh chilt dupc tnet d l vd de ddng han.
Cdn khi su dung NaOH de xt> ly rong, do NaOH Id mgt kilm manh se Idm anh hudng den mgch polysaccharide, cu the Idm cat m@ch lien kit d i n d i n rong bi gay vun, gdy Ihit thodt carrageenan, lam giam higu suaL
2.2. VesCecdong
Carrageenan thu dupc tt> phuong phdp su dung enzyme cd sdc dong cao hon han vi^c su dyng hda chit. Do enzyme c6 tdc dgng nhe nhang vdo thdnh l l bdo rong (hinh 5, 8) nen It dnh hudng den cau trtjc cCia Carrageenan, cdc lien kit glucoside khong b| cat dii't nen sue dong cao. Tuy nhien, khi su dung kilm thi ngoai tdc dgng bdo mdn thdnh t l bao rong, no con tdc dgng vao chuoi polysaccharide (hinh 4, 7) lam cdt dut cdc lien k i t glycoside ndn sire dong cua Carrageenan thip han xu ly bang enzyme.
2.3. Ve pH cCia dung dich Canageenan 1%
Carrageenan thu dupc theo phuang phap XLP ly bang enzyme Viscozyme L cd pH nam trong vCing trung tinh (pH 6,2), trong khi Can-ageenan thu Swfc theo phuang phdp xd 1^ bang NaOH ci5 pH nam trong vung kilm linh (pH 8,6). Qud trinh ngam xii IJ rong vdi NaOH 6% trong thdi gian 40 phut, NaOH khdng chi phd va thdnh le bdo ben ngodi cua thSn rong ma edn c6 thdi gian de tham thiu sSu vdo cac Idp t l bdo bdn trong thdn rong. Do do, sau khi xiS ly xong vdi kiem, mac du quy Irinh da ed cflng dogn ri>a trung tinh rong. Tuy nhidn, cdng dogn nay chi logi bo dupc Idp kiem bam d be mat than rong vS khdng ed tdc dyng vdi cdc Idp ben trong. Din Sk san pham thu dupc van edn Idn hda chat k i ^ . Ngodi ra, chinh vi cdng dogn rua trung tinh khong cd tdc dung vdi cdc Idp bdn trong dan d i n con t6n du NaOH trong cdc Idp t l bao thdn rong, ndn khi nlu chilt k l l hpp vdi nhl#l dO cao ldm cat mgch Carrageenan dan d i n siJc dong cua Canageenan gidm. Ngodi ra eon gay khd khan cho c6ng dogr tinh c h i vd t^ng dyng trong thuc pham do nhiem dl lupng NaOH.
14 • TI5UONG OAI HOC NHA TRANG
Tap chi Khoa hoc - Cong nghe Thuy s So 4/2014
2.4. Ve mdu sSc carrageenan
Khi xu ly vdi kilm ngodi tdc dyng bao mdn than rong cdn phd vd nhilu Idp t l bdo sdc to, khO sdc to trong rong va khd nang t l y trdng nen Carrageenan thu dupe cd mdu trdng sang han. Ngodi ra, mol trudng kiem khi> dupc lipid vd mgt so protein h6a tan trong kilm. Trudng hap XLF ly bang enzyme, trong qua trinh bdo mon thdn rong thi da phd vd mpt phan Idn t l bdo sdc to nhung v i n chua tri^t d l nen sdc to v i n c6n luu Igi mpt it. Do dd, sdn phim Carrageenan thu dupc cd mdu i l l han so vdi dung NaOH trong cOng dilu kidn Igc nhung khdng cheeh lech ddng ke.
IV. K^T LUAN VA KI^N NGHj 1.Kltlu$n
Si> dyng enzyme Viscozyme L xu ly rong cho hi^u suit quy trinh va sue dong carrageenan
cao han, khIc phyc han c h i cua qud trinh sdn xuat carrageenan theo phuang phdp st> dyng NaOH xu ly rong nhu: can-ageenan thu dupe l i n kiem, 6 nhilm moi trudng, dgc hgi cho ngudi sdn xult. Enzyme Viscozyme L c6 ca sd 1;^ thuylt eung nhu thpc tien de thay t h i hda chit xi> ly rong syn Kappaphycus alvarrezii (Doty) Doty trong qud trinh sdn xult Carrageenan.
2. Kien nghj
Tilp tyc nghien cuu thill k l thiet bj d l ca gidi Ida san xuat, hudng den viec thuang mgi hda sdn pham; Nghidn cuu thdnh phln ctJa dung djch hi§n dang bd di sau xt> ly enzyme. T u do, d l xult hudng tj-ng dyng thfch hpp, nhu c h i bien phdn bdn Id trong ndng nghi#p; Sau cong dogn Ipc, m^t lupng bd (xdc rong) tuang d6l idn duac loai ra, cd t h i nghien ct^u c h i biln dung Idm tht>c an chan nudi hodc Idm phdn bdn hCru ea
TAI LIEU THAIVI KHAO Tieng Vift
1. Le Thi Thiiy King, 2012. Nghi6n cihi sir diing Enzyme poiysaccharase dfi thu nhan CarTageenan tur rong si^n Kappaphycus alvarzeii (Doty) Doty Luan van Thac si, Tmdng Dai hpc Nha Trang.
2. Dao Trpng Hieu. 2007. Toi uu hoa quy trinh cong nghe lach chiet Carrageenan tir rong sun Kappaphycus alvarezii. Cue Ch8 bien, Thtrong mai Nong Lam Thiiy san va Nghe muoi, Ha Ngi.
3. TrSn Thi Luyen, 2010. Nghien ciiu xac lap quy trinh san xuat carrageenan ban tinh che tit rong sun Kappaphycus alvarezii Cam Ranh - Khanh Hoa, Tap chi Khoa hpc - Cong nghe Thiiy san, 1,3-10.
4. TrJin Van On (Chu bien), 2012. Thuc tap thyc vat va nhan biSt cSy thu6c D?i hpc Dupc Ha Ngi, Ha N$i.
TlingAnh
5. Craigie J.C, Leigh C, 1978 Carrageenan and agar. In handbook of phycoligical method. Physiological and Biochemical methods - Cambndge Univ. Press, 9 - 3 1 .
6. Soovendran A/I Varadarajan, Nazaruddin Ramli, Arbakanya Ariff, Mamot Said, Suhaimi Md Yasir, 2009 Development of high yieldmg carragenan extraction method from Eucheuma Coionii using cellulase and Aspergillus niger, Prosiding Seminar Kimia Bersama UKM-ITB Vm - 461.
TRUONG OAI HOC NHA TRANG • 15
