NGHIEN CUU SlTDU^dNG HOA CARBOHYDRATE CUA RONG NAI SARGASSUM BANG AXIT
STUDY OS SACCHARIFICATION OF C.^RB0HnR4TE OF BROWN SE WEED SARGASSUM BY ACID TREATMEST
Le Thi Tirdng' Ngiy rt,i„bk.22/M015, Ngi, phi, I . i „ , h 6 . „ „ ,0,6701!: Noi, taj, d i . , Ivj.^oie TOIM TAT
"'^'«'''"''''S'ti^''cdnnd,-nhdm,icdinhioa,aeidyiencdiiakiendi,dngh6acarboh,dratecnarong
» » . ^ ^ ™ , „ ; v c v ™ , „ i „ „ j „ ( U ™ c„„c*„ „ s * ^
rong bien Ket qua nghien cua cho thiy dirdng hoa carbohydrate cno rong nin Sargassun, poiycyslnm hdng
hoe carbohydrate tn Sargassam p„i,eyslun, bing a,yii sun/uric ii f - I2(rc, ning dd aw sunfnnc ' %y ,i
"••" Sinn thiy phin 12t)phal tha nhin him Inong dirong eao nhil la 47,3Jnig/.Sg min i„ng kho "
Tit khoa: Rong bien, rong niu Sargassam polycyslam, m l sanfiiric ABSTRACT
The purpose of this study was to determ.ne whal kind of ae.d and saecharification eonditions of
bio-ethonol from carbohydrate of brown algae The results showed thac the efficiency of earholiydrje
« c * „ y i « of Sargassam polycy-slam by sulfuric acid Irearme.n was ovo folder higher than Hat by '^l»''>'''^'''t''-'''>,'ntSa,,able parameters to sacchanfy. carbohydrate of Sargassam poLsiun, by,nlfanc aeul were ach,eved a, die lempera.ure, the acid com-entration and the duration were 1 VC rf. volnme 121, minntes, respectively The highe.l sugar w-as 47,33mg,,gram algae ,in dried weight,
Keywonls: Seew-eed Sargassum poiycyslnm. sulfuric acid
I. OAT V A N Bt
Hl^n nay, san lu'p'ng rang bien tren the gim
«9t Ithoang 7.000.000 t i n rang turn thiFung phim/nam, trong do hon 50% san lu'ong la do nu6i trong. tSp tmng 4 cac nu'*c Trung Quoc.
Han Quoc vS Nhjt Ban [7]. Cdn o Viet Nam.
sSn luvng rong bien cung co the khai th^c len il4n 79.126.3 tan rang kho/nSm. RISng tai khu VU'C Vinh Nha Trang. KhSnh Hoa trO lirong rong n^u len i3en 4840,4 tan kh6/nam, rong do
1^ 231,97 tan kho/nam v i rong luc la 16,53 tan kh6/naml1,2].
Rong blln du'oc xem l i mot nguon nguygn lieu doi dao, re va co gia tri. Ngoai viec si> dung rong bien lam thijc an cho gia sijc, gia c i m nhu tn/oc dSy thi hien nay cac nghien ciiu chiet njt va irng dung cac chat co hogt tinh Sinh hoc t i l rong bien trong y duoc va thuc pham kha ttianh cSng o nhieu nuoc tren th4 gioi trong dd cd Viet Nam. Va nghien cftu san xuat
' -ThS. L« ThI Tudng: Khoa C « n , n g h j Biire p h i m . Tnrtng O j i hoc N h , Trang
TRUONG OAI HOC NHA TRANG . 6
Tgp chi Klioa lioc - Cdng nghe Thuy smi ethanol smh hoc lu nguon rong biin duac xem la mpt hudng nghien CLCU mdi. Mac du hien nay con nhiiu tranh cai v i tinh hieu qua ciia no song nhiiu nha khoa hoc da khing dinh ring, rong bien duac xem nhu mot nguin nguyen lieu mdi, thay the mot phin cac cay luang thuc de san xuit ethanol smh hoc cho tuang lai [7,9,11,12,14,15,16,18]
Tuy nhien, 6e co ca chat cho qua trinh len men san xuit ethanol sinh hoc tu rong bien can phai co qua trinh dudng hoa carbohydrate Id rong biin [19], lam ca chit cho qua trinh len men, Hien nay tren t h i gidi da co nhiiu cong trinh nghten cuu dudng hoa carbohydrate tu rong biin bang nhiiu phuang phap khac nhau nhu phuong phap hoa hoc (dung axiE, kiem), phuong phap enzyme Trong dd, phuang phap dung axit dudng hoa carbohydrate tu rong bien duac danh gia la hieu qua nhit [10.13)
II DOI TLfONG VA PHU'aNG PHAP NGHIEN GUI' 1. Doi tirp-ng, vat lieu nghien ciju
Rong nau Sargassum mcclure (S mcclurej.
Sargassum polycystum (S polycystum).
Sargassum microcystum (S microcystum).
Sargassum binden (S binderi) duac mua tai
So 1/2016 cac dau ngu tu Nha Trang, Van Gia va Ninh Hoa. Rong da dupe phai kho va chua trong cac tui polypropylen, duac bao quan d nai kho rao, thoang mat. Sau dd van chuyen ve phong thi nghiem diJng dan.
Acid sunfuric (H^SOJ. acid ascorbic (CfiHgOJ, sat (III) sunfat (Fe,(S0J3), kal.naln tartrat (C^H^O.KNa. .4H,0), xut (NaOH), acid acetic (CH3COOH), natn acetat (CH^COONa), ding sunfat (CuS0^.5H,0). Tat ca cac hoa chat tren co nguon goc tu Merck - Due, dupe sd dyng pho biin trong phan tich hda hoc.
2. Phu'O'ng phap nghien CIFU 2 1. Cach lay mSu va chuin bi miu
Cach tay mau: Rong nau duac thu mua ti>
cac dau nau tai Nha Trang, Van Gia va Ninh Hoa Mii vung thu mua ngiu nhien 50kg rong nau kho. Rong da duac phai kho va bao gbi trong cac tui polypropylen, dupe bao quan d nai kho rao, thoang mat Sau dd van chuyin ve phdng thi nghiem
Chuan bi m i u : Rong nau kh6 dupe phan Ioai, lam sach, sau dd xay nho roi cat giij' trong cac till polyetylen Rong duac bao qu^n d nhiet dp thudng, nai kho rao. thoang mat M6i thi nghiem dung 5g rong kho da xay nho.
^'1 • mu'ONG DAI HOC NHA TPANG
ISEShilOmaJioc - Ciing mjhe TImy san 2 2, Quy trinh nghien ciru ting guat
Rong nau — • Lam sach, phan loai khd
Aacoinn nicumy ^ ^ , „ . .
" " " " a phan " ^ - "
Bo tri thf nghiem.
Xac dinh loai rong nau ohira ham lirong carbohydrate cao nhit. Sir dung 4 loai rong nau kho Sargassum mcclure (S.mcclure).
Sargassum polycystum (S.polycystum), Sargassum microcystum (S.microcystum), Sargassum binderi (S bmderi) da xay nhd, mdi mau Ihi nghiem la 5g Xac dinh ham lirong carbohydrate cua rong theo phuong phap gan dung cua Nguyen, 2011 [21]
Xac dinh loai acid thich hop de dudng hda carbohydrate cua rong nau- l«6i mau thi nghl6m can 5g rong nau khd da duoc xir ly, xay nhd, cho vao binh tam giac 250ml Sau dd chd them 100ml nudc cat, tien hanh boc kin binh, dem thuy phan o nhiet dp 110«C, thdi gian 100 phut vdi CiSc m j u nhu sau- m i u 1 (khdng bo sung acid), mJu 2 ( bo sung 1% acid sunfuric), mau 3 (bo sung 1% add ascorbic) Tiep dd loc loai ba va xac dmh ham luong dudng khir cua cac miu dich loc b^ng phuong phap Bertrand [3] Tip dd sd sanh. lira chdn loai acid thich hop cho dudng hda carbohydrate cua rong nau
Xac dinh cac dieu kien du'dng hda carbohydrate thich hop bJng acid sunfuno IMdi mau thi nghiem can Sg rang nau khd da duoc xii- \)/, xay nho, cho vao binh tam giac 250ml Sau dd cho them 100ml nudc cat, bd sung acid sunfunc d cac neng dp 0 5%; 1%, 2%; 3%
4%; 5%; 6%, tien hanh bee kin binh, dem thuy phan d nhiet dd 110"C, thoi gian 100 phut Tiep do ipc loai ba va xac dmh ham luong dudng khu' cua cac m i u d|Ch loc bing phuong phap Bertrand Tu do lira chon ndng dp acid thich hop cho qua trinh du'dng hda carbohydrate.
Xay nhd Xac dinh ham luong carbohydrate
Thuy phan bing . acid
- Loai acid - Ndng dd acid - Nhiet dd thuy phan t - Thdi gian thuy phan Bd tri thf nghiem tirong tu ddi vol nhiet dp va thdi gian thuy phan, vol dai nhiet dp va Ihdi gian thijy phan i i n luot la 100"C, 105"C, 110"C, 115"C; 120"C, 125«C va thdi gian 60 phut, 90 phlit, 120 phut, 150 phut va 180 phut 2 3, Phuong phap phan tich
Xac dmh ham luong carbohydrate ciJa rong bien theo phuong phap gan dung (Nguyen, 2011) Carbohydrat (%) = 100% - (Ham iuong arn-r iipit thd * protein thd y tro) Xac dinh lirong dudng khu- then phuong phiSp Bertrand XiSc dmh thanh phin ciSc loai dudng bing sac ky trap doi ion hieu nang cao tren miSy GC- FID Agilent (My) HP3ICS-3000. cdt phan cue nhe. SP 17 A, d i u do PID Xac d|nh khoi luong cac mau rang bing can phan tich cd dd chinh XiSc lO^g, Shimadzu Nhat XiSc dmh dd am cua rcng thed TCVN 3700-90 Xac dmh ham iuong tro cua rong theo AOAC 938 08 Xac dinh ham luong protein thd theo TCVN 3705- 90. XiSc dmh chit beo thd bing phu'ong phiSp Folch theo nguyen t i c diing hdn hop dung mdi Chloroform.Methanol vdi ti le 2 1 de hda tan tat CiS chat beo trong rong, liSch Idp va chiet qua pheu loo nhiiu l i n Sau khi liSm bay hoi hdt dung moi, can chat b<io cdn lai via tinh ra hiSm lu'ong hpit the trong lOOg rong
2 4 Phuvng phap xCr ly s6 heu
Tai CI3 cac thi nghiem deu duoc [lin hanh lap lai 3 lan So tieu duoc xu iy thong ke ANOVA de biit so khac biel cd y nghia thdng ke giua CiSc giiS tn trung binh vdi a = 0,05% viS Post Hoc Test sau ANOVA d l bidl cu the su khiSc biet cd y nghTa thdng ke giua cap gia tn Irung binh bing phan m i m SPSS 16 0
TRUONG OAI HOC NHA TRANG . 71
Tup L hi Khoa hoc - Cdng nghe Thuv
1 Kit qua xac dinh ham lu^ng carbohydrate cua rong nau khai thac tai cac viing bien Khanh Hda Bang 1. Ham lu'p'ng carbohydrate cua 4 loai rong nau khai thac tai
cac viing biin cua Khanh Hda {% rong kho tuyet doi)
Ten rong nau 1 S bmden 1 S mcclurei
S microcystum 1 S polycystum
1 Thanh phan 1 Carbohydrate (%) 1 Carbohydrate (%) 1 Carbohydrate (%) 1 Carbohydrate (%) Ham luong carbohydrate trung binh
Nha Trang 53,9^
54,9^"
ss.S"- 67,2^"
:ua rong nau Ninh H6a
54,5'=
57,2""
56,4^- 68,3"'
Vgn Gia 53,6=
55,2^'=' 56,4"' 68,2''' khai thac tai Khanh Hoa
Gia tri trung 54^
55,8»
56,2"
67,9=
58 5 Ghi chu ChO
Nhan xef va thao luan kit qua- Ket qua phan tich the hien 6 bang 1 cho thay, ham luang carbohydrate cua 4 loai rong nau khai thac tai 3 vung biin Khanh Hoa dao dong kha cao, tu 53,9% d i n 68,3% Tuy nhien, ham luung carbohydrate trong mpi loai rong nau khong co su dao ddng lan giii'a cac vung mien
Ham luang carbohydrate trung binh cua 4 loai rong nau khai thac tai Khanh Hoa la 58,5%.
trong do cao nhat la S polycystum (67,9%) va thap nhaiiaS bmden (54%) K i l qua nay cung
su khac tjiet co y nghTa thdng ke giim c$p gia In trung binti phu hap vai ket qua nghien cuu nhom tac gia cua Vfen hai duang hoc Nha Trang khi khao sat ham luang carbohydrate trong rong nau tai mot s6 vung biin cua Viet Nam la 55%- 62% [5] Tuy nhien, khi so sanh ham luang carbonhydrate trung binh cua rong nau tai vung bien Ireland thi ham lugng carbohydrate trung binh cua rong nau tai vung bien Khanh Hoa thap ban 2% (Reith va cong su 2009).
2. Ket qua xac djnh loai acid dung du'dng h6a carbohydrate cua rong nau S.polycystum
1 1
l l i n l i 2 Kha nang dirimi; hiia i i i r b u l n d r a l i - ci 1,1 l,„ ( „ , flai loai arid kiiiic ntiai Acid sunfunc va acid ascorbic la nhu'ng acid duoc dung kha ph6 bien trong nghien cuu thuy phan do nang luc hoal hoa cQa nd khiS cao va It tac dung oxy hoa manh |4] khi so siSnh cung nhom acid vo co hay huu co Ngoai ra, nhieu kel qua nghien cuu ngoai nu-o-c khang dmh,
^ : • I-IJ-O-NG DAI HOC .NHA IRANG
liln nau S,polye\
acid sunfuncla acid dai dien chonhdm acid voce ViS acid asccrbic la acid dai dien cho nhom acid hu-u CO CO khiS nang thuy phan carbohydrate rong bien cho hieu qua cao [10,13] Vi vay tac giiS chpn hai acid niSy de khiSo sat quiS tnnh dudng hda carbohydrate cua rong nau
S polycystum Ket quia thu diroc Old hien d hmh 2 cho ihiSy m i u duong hda carbohydrate bang acid sunfunc cd hiSm lirong dudng khir tao ra giSn gap ddi so voi m i u dodng hda carbohydrate bang acid asbortiic viS cao hon gap 10 lan so voi m i u ddi chdng. Dieu nay chdng td, acid ascorbic vi, acid sunfunc deu cd kha nang duong hda carbohydrate cua rong nau S.polycystum, luy nhien sir dung acid sunfuric cho hieu qua dudng hda cao hon nhieu so vdi acid ascorbic
Muc dich chinh ciia quiS trinh dudng hda carbohydrate nhim c i t dul cac lien ket glucdcid trong hop chit carbohydrate d i tao thiSnh ciSc
monosacchande hda tan. ddng thdi cung nham tao dieu kien IhuiSn loi d i cac dirong hda fan ben trong ciia rong khuech tan vao dich Ihuy phan nhanh hon Tuy nhien muc dd c i t dul CiSc hen ket phu thudc vao tiing loai acid khac nhau. acid vd co cd nang luc hoal hda cao hon so vdi acid hCru co nen muc dd cat dul CiSc lien k i l cung cao hon. K§1 quii niSy dung vdi ket quiS nghien cuu ciia Leilei Ge vis cong su 2011
3. Ket quiS XiSc dinh diiu kien dyd'ng hda carbohydrate ciia rong S,polycystum bang acid sunfuric
3 1 Kk qua xac dmh ndng do acid
Nhan xet va thiSo luan kit qua Ti> ket quiS phan tich t h i hien d hmh 3 cho thay hiSm luong duong khu tao thanh ciia qua trinh Ihdy phan phu thudc nhieu vao ndng dp acid sunfuric Khi ndng dd acid sunfunc tang tir 0.5% den 3% Ihl hiSm lirong dudng khir cung tang va dat cue dai tai ndng dd 3% 135,67 mgl nhung lai khdng khiSc biel Cd y nghia v i mat thdng ke so vdi ndng dd acid sunfunc 2S i34,4mg) Neu tiep tuc tang ndng dd acid sunfunc hon 3% thi ham lupng duong khu lai giiSm VI vay, ndng dp acid sunfunc 2-: duoc xem la thich hop cho
qua tnnh thiiy phan carbohydrate tao thiSnh dudng khii-
QuiS tnnh thuy phiSn rong biin bang acid la do su xuc tac ciia nhiet dd viS acid trong mdt thai gian nhit oinh lam cac hen k i l trong ciSc hop phin cao phan tu bi phan o i l sau dd siSn phim thuy phan duoc tiSch ra khdi co chat va khuech tan vao dung dich Hieu suilt thuy phan phu thudc kha nhieu vao ndng do acid Ndng dd acid thap thi hieu suat thuy phan khdng cao.
tuy nhien ndng dd acid qua cao se pha huy biSn chat ciia cac duong lam suy giam ham luong duong trong d'Ch thuy phan [6]
~-'C"-iG D^] H^C NHA TPANG • -•
Tap c hi Khoa hen - Cong nghe Thuy san 3 2 Ket qua xac dmh nhiet do thuy phan
Nhan xet va thao luan kit qua Ket qua phan tich t h i hien a hinh 4 cho thay, nhiet do anh huang dang ke den hieu qua thuy phan carbohydrate bing acid sunfunc Cu the, khi tang nhiet do tu' 100°C den 120^C thi ham luang duang khic tao ra tang lin luat tu 22,7mg/5g mau len d i n 41,5mg/5g miu, nhung khi nhiet do thuy phan tang d i n 125*^^ thi ham luang duang khu lai giam Diiu nay cd t h i giai thich, nhiet do thiiy phan cang cao thi cac t i bao rong nau gian na cang nhieu, su khuich tan acid vao cac lap ben trong nhanh ho'n, hieu qua thiiy phan cao ban, cac lien ket cao phan tu se nhanh chong bl be gay Nhiet do thuy phan thap Ihi kha nang luong tac giua co chat va chit xuc tac
yeu han, kha nang tuang tac len cac hen kit cao phan tu cung kem ban, nen phan ung thuy phan diin ra cham han, do do ham luvng duang khu tao ra it ban. Tuy nhien, khi nhi^t do qua cao thi mot phan cac duang dan se b|
pha hiiy trong moi truang acid, vi vay cd sg' giam ham lugng duang khu khi tang nhiet 6Q thuy phan Do do, nhiet do 120''C duac xem la thich hgp de ihuy phan carbohydrate cua rong nau S polycystum. K i t qua nay phu hap VO'I DuBok Choi va cong su, 2009; Leilei Ge va cdng su, 2011 khi cac nhom tac nay deu cho ring nhiet do thiiy phan carbohydrate tcr rong bien bang acid sunfunc phii hgp nhat d 121''C trong thai gian til' 1-3h
3.3 Kit qua xac dmh that gian thuy phan
iRUONG DA) HOC NHA TRANG
- Cong nghe Thuy san Si I 21116 Nhan xet viS IhiSo luan kdt quiS-
_ Ket qua phan tich the hien d hinh 5 cho thay thdi gian thuy phan cd iSnh hu-dng d i n ham luong dudng khip tao thanh. Luong duong khi, tang khi thdi gian thuy phan tiSng tu 60 phtit den 120 phut. Tuy nhien, khi h i p tuc tang thdi gian thuy phan thi hiSm lirong dudng khil tad ra lai giam. Diiu niSy cd the giai thich ring thdi gian thuy phan cang diSi thi cang lam tang kha nang tiip xuc cua acid vol CO chit, tad dieu kien cho cac SiSn phim thuy phan khuich tiSn vao dung dich. Neu thdi gian thuy phan qua ngin thi phiSn irng dien ra
chua tnet de. su tiip xuc giua acid viS co chit quiS ngin nen chua du luc de pha vd va c i t dut CiSc lien k i t trong phan til- polysaccharide, tdc do thuy phan cham, do dd ham luong duong khu- tao thiSnh khdng cad. Tuy nhien, niu thuy phan trong thdi gian diSi (hon 2 gid) d nhiet dp cao (120-'C) trong mdi tru'dng acid thi mdt sd Idai du-dng sau khi tao ra tti- quia trinh thuy phan se bi phiS buy [6] nen ludng dudng khir thu duoc cung se khdng cad
4. XiSc dinh cu the cac loai dyd'ng trong dich du'trng hda rong nau S,polycystum Bang 2. Kit qua xac djnh cy t h i cac loai du.d.ng trong dich du'dng
hoa rong nau S,polycysturn
Ket quiS phan tich sac ky cho thay trong 5g S.polycystum nguyen lieu sau khi dudng hda bing acid sunfunc d dieu kien nhiet dd 120°C.
ndng do acid 2%v, thdi gian thuy phan 120 phut chd hiSm iuong dudng manitol cao nhit (24%), Hep theo liS fructose (13,3%), galactose (9%).
Nhdm cd hiSm luong tuong ddi thip la xylose 1,7% va mantose 1,25% va glucose (0,72%)
••• KEiT LUAN VA KIEN NGir 1. Ket luan
1.1. Ham luong caibohydrate cua rong nau S,polycystum la cao nhit, chiim 67,9%, trong khi S bmden, S mcclure, S.microcystum lin luot la 54%, 55,8% va 56,2%.
1.2. Ciing khoi lu'ong m i u 5gram rong kho.
ciing dieu kien thuy phan d nhiet dd 110^C, thdi gian 100 phut nhung acid sunfuric da t h i hien kha nang thuy phan carbohydrate cua rong nau S,potycystum cao g i n g i p ddi so vdi dung
acid ascorbic, ham luong dudng khu lin lu'dt liS 29mg va 16,9mg
1.3 Oudng hda carbdhydrate cua rong nau S polycystum bing acid tot nhat d diiu kien nong dd acid sunfunc 2%v nhiet dd 120°C va Ihdi gian 120 phiit Kit quia thu duoc hiSm lu'ong dudng khi> cao nhal la 47,33mg/5gram rong khd, trong dd manitol chiim 24%, fructose.
13.3%, galactose: 9%, xylose 1,7%, mantose:
1,25% va glucose 0,72%
2. Kien nghi
De tang hieu quiS dudng hda carbohydrate cua rong nau S.polycystum. c i n flip tuc nghien cipu qua trinh nin xii- ly rong bang acid sau dd thuy phiSn bing enzyme, tren co sd dd danh giiS, lua chon phuong phiSp dudng hda carbdhydrate cua rong nau S polycystum hicu quiS nhat, liSm co sd cho nghien cuu san xuit ethanol smh hoc tir- carbohydrate cua rong blin sau nay.
TRUONG OAI HOC NHA TRANG . 75
Tap chi Khaa Ink - Cong ,ighc Thu\ san
TAI I IKl T H \ M K H \ 0 Tiing I let
Li Nhu Hau ^ a coiia -ir COOOl De lai • N^hicn cmi \ a di- vi.al giai pliap khai chac hop Iv va K-n \ imi! cho roni;
nau}Cn heu san \ u a i clhanol ci \ c n bicn M i a Trang"". Ti 1-23
Lc Miir Hau \ a cong six CO lOi Tiem nang rong bien lam nguyen lieu san \ u a t ethanoi nhien lieu lai Viei Nam
\^M cao h6i nghi Khoa lioc k\ mem 3> nam \'ien Khoa hoc va Cong nghi' Viet Nam - |-la Noi Tr 260-265 Lc Thanh Mai va cong su (20041 Cac phtrong phap phan tich nganh cong nghe len men, N X B Klioa hoc \a kv ihuat Tr 52-54
Tran Thi Luvcn \ a cong sir. (20001 Cac phan irng co ban va bifin doi cna ihirc pham trong qua trinh c o n - n^^hc
\ X R Niing nghiep T r 2 - J 6 '^ ""
Tran Thi LuNtJn. Do Mmh Phung Nguyin Anh Tuan, Ngo Dang Ngliia. (2004) Che bien roni; bien N X B Noni;
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Le Ngoc Tu va cong str. ( I 9 ' J " | . Hoa Mnh hoc cong nghit;p N X B Khoa hoc va k\ ihuJI Tr 2 4 9 - 2 M
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" l ! r ' ? ' " V ' r > " ' ' ' ' ' " ' ^'•'"^•'' ^ ' ^ ' ^ • ^ > ^ ^ ^ ' - ' " " ' - Toshilsugu Sakou, (2007) Seaweed Bioelhanol Pmduciion in Japan I he Ocean Sumrisc Proieci
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