KHOA HOC C 6 N G N G H l
AIMH HLTOIMG CLIA T H O I GIAIM T H I J Y PHAIXI V A L O A I EIXIZYIVIE DEIM Q U A T R I I V H T H U Y P H A I M SLJIM CA M A P
[Carcharhinus dusmumierii B A I \ I G PROTEASE Dinh Hfti Dong^, VQ Ngpc BOi^ Nguyin Thi My Trang^
T6MTAT ^ Bai bao n^y trinh bay mot phan ket qua nghidn ctiu thuy phan sun ca map {Carcharhinus dussumieri) bdng
enzyme protease. Ket qua nghi&n ciiu cho thdy cac enzyme: neutrase, alcalase, flavour2yme, papain va h6n hgp enzym alcalase - papain deu co kha nSng thuy phan h6n hop sun ck mgp cr dilu ki^n pH tu nhi^n, ty le enzyme bo sung 0,2%, luong niroc bd sung 100% va nhi^t d6 thuy phan 50"C. Trong dieu ki^n tren, h6n hpp enzym alcalase - papain co kha nang thiiy phan sun ca map tao thanh lugng protein hoa tan, peptid, aad amin, chodroitin sulphate cao hon nhilu lan so voi viec thuy phan sun ca map bdng enzyme protease khac da thit nghidm.
Ti khda: Neutrase, alcalase, Qavourzyme, papain, chodroitin sulphate, sun ca map, thuypbin.
LMaOAU
Chondroitin sulphat Id thdnh phdn co ban cau tao ndn sun khdp vd cau tao ndn cdc td chirc soi chim (gan, co, day chdng...) giup cho su vdn dpng hnh ho^t vd tinh dan hoi trong hoat dpng khdp, tao do bdn khi bi nen ep. Chondroitin sulphat lam tang san xuat chat nhay vd kha nSng bdi tron cua dich khdp, dam bao chuc nang dinh duong va su van dpng linh boat ciia khdp. Ti vay, chondroitin sulphat duoc su dung de hd trp dieu tri cdc benh ly ve xuong khdp, ban chd qua trinh thodi hod khdp. Chondroitin sulphat con co vai tro bao ve sun khdp bdng each uc che cac enzyme phd huy sun khdp nhu coUagenase, phosphohpase A2, N-acetylglucosamindase. Ngoai ra, chondroitin sulphat cflng gop phan nudi dirong vd tdi tao cdc td bdo cua gidc miic mdt [5, 7].
Hidn chondroitin sulphat chii yeu dupe thu nhan hi cac ngudn nguydn hdu hj nhien. Trong do, sun cd md dac biet la sun ca map thuong co hdm lupng chondroitm sulphat cao hon cdc loai dpng vat khdc.
Chinh vi the, ngirdi ta thudng su dung sun h-ong san xuat chondroitin sulphat dung trong san xuat thuc pham chuc nang hd tro dieu tri cdc b^nh xuong khdp. Chondroitm sulphat thuong hen ket voi protem bdng Uen ket o-glycosid tao thanh mpt proteoglycan (PG) (glucoprotein) ndm trong cau tnic cua md sun
nen con nguoi rat kho hap thu [5, 7]. Hidn cd mdt so ngbifen ciru sir dimg tdc nhan hoa hpc hodc enzyme thuy phan sun cd khd hoac tuoi vd kdt tiia thu chondroitin sulphat dung trong san xuat thuc phdm chiic nang [5, 7]. Tuy vdy, viec chi thu nhdn chondroitin sulphat ddn den su lang phi ngudn cdc chat tu nhien hi sun ca vd hieu qua su dung khdng cao ciing nhu hidu qua san xudt han chd, gid thdnh san pham dat [5, 7], De khdc phuc hien tuong nay, da hdn hdnh nghien cuu su dung enzyme trong thuy phan sun ca map tuoi vdi mong mudn tao ra djch thiiy phan chiia cac chat tu sun ca vd chondroitin sulphat de dinh huong sii dung 1dm thuc pham cung cap cdc thdnh phdn thiet yeu giup ho trp chdng thoai hoa xuong khdp. Trong gioi han ctia bdi bdo ndy, chi cong bo phan nghien cihi lua chpn enzyme trong thuy phan sun cd map.
2 . NBUYBU VAT UEU VA PHUDNG PHH> NGHBII CUU 2.1. Nguyen vdt h$u
2.1.1. Sun camap
Cd mdp trdng {Carcharhinus dussumien (Muller
& Henle, 1839)) dupe thu mua nguydn con tai cdc tdu khai thdc t^i vung bien Khanh Hoa. Sau thu mua, ftu todn bp vdy cd, sun ca vd van chuydn ve phong thi nghiem. Tai phong thi nghiem, tien hanh xir ly loai bo thit, mo hen kdt, 1dm sach, cap dong va bao quan dong 6 -20*'C dd dung trong sudt qua tiinh nghidn
^ Truong D^i hpc C6ng ngliiep Thirc phdm TP. HCM ' Tniong D^ hpc Nha Trang
Email: [email protected] 96
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Hinh 1. Hinh dnh v^ vi sun cd m^p {Carvharhinus dussumieri) sau khi xii ^ 2.1.2. Enzym alcalase
Enzym alcalase 2.4L la chd pham protease thuong mai do hang Novozyme - Dan Mach cung cap. Alcalase thudc nhdm enzyme serine endopeptidase co cdc dac tinh ky thudt nhu; pH thich hpp trong khoang 6,0 - 8,0; nhi^t dp thich hpp 30 - 65''C, hogt tinh 2,4 AU/g dupe bao quan 6 0 - 5"C.
2.1.3. Enzym neutrase
Enzym neutrase 0.8L Id chd pham protease thuong mai do Cong ty Advanced enzyme Technologies Ltd - India san xuat va cung cap tai Vidt Nam. Neutrase thupc nhom enzyme endopeptidase CO cdc d§c tinh ky thuat nhir pH thich hpp 5,5 - 7,5, nhidt dp thich hpp 40 - 50*^, boat tinh 0,8 AU/g dupe bdo quan 60-5°C.
2.1.4. Enzyme Flavourzyme
Flavourzyme la che pham protease thuong mai do hang Novozyme Dan Mach cung cap.
Flavourzyme co ca boat tinh cua endopeptidase vd cua exopeptidase nhung chu yeu la exopeptidase, cd ngudn gdc hi Aspergillus oryzae [4], boat dp la 500 lAPU (Leucine Aminopeptidase Units)/g, dieu kien hoat dpng thich hpp Id 50 - 55°C, pH = 5,0 - 7,0.
2.1.5. Enzym papain:
Papain thuong mai cd boat tinh > 2,0 mAnsonU/mg (ca chat hemoglobme, pH 6, nhiet dp 35,5°C) do Merck - Diic san xuat Papain la mpt enzyme chiu dupe nhidt dp tuong ddi cao. O dang nhua khd, papain khdng bi bidn tinh trong 3 gio b 100°C, con b dang dung dich, papain bi mdt boat tinh sau 30 phiit b 82,5''C. Papain cd pH thich hpp 4,5 - 8,5, d§ bi bien tinh 6 pH<4,5 vd b pH >12.
2.2. Phuong phdp nghlto ciiu 2.2.1. Cic phuong phip phin tich
- Dinh lupng ham lupng nito axit amin (Naa) theo phuong phdp formol; dinh lirpng NH3 theo phuong phap chung cat loi cudn hoi nude [6].
*Jac dinh hoat dp protease tiieo phuong phip Anson [21
Nguyen tdc cua phuong phdp: dimg - protein casein lam co chat xac dinh boat dp thuy phan protein cua enzyme protease tren co sd dinh lupng san pham dupe tao thanh trong phan ung hdng phan ling mau vdi thudc thir FoHn - Ciocalteau. Dua vao do thi chuan tyrosine de dinh lupng tuong ling vol lupng san pham tao thdnh dudi tdc dung cua enzyme.
- Dinh lupng protein hoa tan theo phuang phap LowrylS, 91:
Nguyen tac cua phuong phdp Id cdc axit amin co vong thom, Tyr vd Trp co mat trong protein se phan img vol thudc thir Folin-Ciocalteau tap thanh phiic chat mdu xanh den co dp hap thu cue dai 6 budc song 650 nm. Dira vao ducmg chuan protem ngudi ta CO the dinh lupng hdm lupng protem.
- Xac dmh him lupng peptid [6, 10]; hdm lupng pephd dupe dinh lupng dua vdo dirong chuan tyrosine. Lay 1 g mdu thiiy phan, cho them 9 rid nude cdt sau dd khudy deu trong khoang 5 -10 phiit roi ly tam lay dich trong de xac dinh hdm lupng peptid nhu sau: lay 2 dng nghiem sach 1 dng thi nghidm va mot dng ddi chiing. Ong thi nghidm: hut chinh xde 2ml dung dich lpc 6 tr^n cong vdi 2 ml Trichloacetic acid (TCA> 20% de 30 phiit rdi lpc qua giay lpc thu dich lpc. Lay mpt dng nghiem sach cho vdo 1 ml dich lpc -(- 5 ml dung dich Na2C03 0,4 M lac deu, roi cho vao 1 ml Folin de 20 phut so mdu b buoc song 660 nm. Ong doi chiing: lay 1ml dung dich TCA 10% -1- 5 ml Na2C03 0,4 M -t- 1 ml folin de 20 phiit dem so mau.
Tinh ket qua: dua vao duong chuan dd tinh luong tysosin tirong ling.
Ham lupng peptid dupe tinh theo cong thiic:
2.2.2. Phuang phap dinh lupng chondroitin sulfate (CS) bing phuong phap so mau theo Famdale va cdng su [9J
Nguyen ly: Dua tr^n su thay ddi trong quang pho h ^ thu cua DMMB (1,9 Dhnethyhnethylene) khi tac dung vdi chondroitin sul&te (glycosammoglycan sulfate) b bude song 525 nm.
Dua vdo duong chuan cua chondroitin sulfate A {gdc 97
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sulfate gan o vi tri C 4 (chondroitin-4-sulfate), CS4) voi DMMB de xac dinh ham luong chondroitin sulfate. Phuong phap nay co dp nhay cao, co th^ dinh tinh va dinh lupng ham lupng CS b muc \ig.
2.2.3. Phuong phip bo tri thi nghiem B6 tri thi nghiem d^ lua chpn enzyme protease thuy phan sun ca map trang dupe trinh bdy b hinh 2.
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Hinh 2. So dd thi nghidm lua chpn l o ^ enzyme proetase thich hpp cho thuy phSn sun ca map
Vi vd sun cd map da xii ly duoc ra ddng, nra va xay nho bang may xay va dupe su dung d^ lam nguydn lieu nghifen ciiu thuy phan bdng cac enzyme protease: neutrase, flavourzyme, alcalase, papain.
Tidn hdnh 5 thi nghiem thuy phdn bdn hpp sun ca map bang cdc enzyme protease khdc nhau: mdu 1 thuy phan bdng enzyme neutrase, mdu 2: thuy phdn bdng enzyme alcalase, mdu 3: thiiy phan bdng enzyme flavourzyme, mdu 4: thuy phan bdng enzyme papain vd mdu 5: thiiy phan bang hdn hpp enzyme alcalase va papain theo ty le 1/1. Cac mdu tiiuy phan d^u su dung 2 kg hdn hop sun ca map, ty le enzyme su dung la 0,2% vd lupng nude bd sung Id 2 ht, pH thiiy phdn la pH tu nhien cua bdn hop sun ca map (pH 6,8) vd nhiet dp thuy phan 50°C. Sau cac khodng thdi gian: 0 gid, 2 gid, 4 gid, 6 gio, 8 gid va 10 gid, tien hdnh lay mdu dich thuy phan de ddnh gid ham lupng protein hoa tan, ham lupng peptid, ham lupng Nan, NNH3 ^3 hdm lupng chondroitin sulphate tao thanh. Kdt qua ddnh gia la co sd de lira chpn enzyme cho qua tiinh thuy phan.
2.3. Tliidt hi va hda chdt
- Thiet hi: su dung cac thiet hi hien cd tai Trung tdm Tlii nghiem Thuc hdnh - Trucmg Dai hpc Nha Trang vd Trudng Dai hpc Cdng nghiep Thuc pham - TP. HCM: May so mdu UV-VIS DR6000 - Hach
(My); bd on nhiet MEMMERT WNB14 - Diic; may ly tdm lanh tdc dp cao HERMLE Z36HK - Diic; he
thdng phan tich hdm lupng nito/protein theo phuong phap Dumas (Due); bon nuoc dieu nhiet Memmert WNB22 Ouc); ndi thuy phan dung tich 30 ht (Vi^t Nam),...
- Cac hda chat su dung ti-ong thi nghiem d^u Id boa chat tinh khiet do Hang Merck - Diic cung cap.
2.4. Phuong phdp xii 1;^ s6 l i ^
Mdi tbi nghiem deu tidn hanh lap lai 3 lan dOc lap va sd Ueu la ket qua trung bmh ciia cdc lan thi nghiem. Kiem tra su khdc biet giiia cac sd heu thdng ke hdng phdn mem Statgraphies Centurion XVII trial.
3. KFT QUA NGHBU CUU VA IHAO LUAN
3.1. Anh huong cua thdi gian thiiy phdn vk lo^i enzyme protease toi ham lugng protein hda tan
Tien hanh 5 mdu thi nghidm thuy phan hdn hpp sun cd map bdng 5 loai enzyme: neutrase, flavourzyme, alcalase, papain va hdn hpp enzym alcalase - papain va sau cac khoang thdi gian; 0 gid, 2 gid, 4 gid, 6 gid, 8 gid va 10 gid, tien hdnh lay mdu dich thuy phan de ddnh gid hdm lupng protem hoa tan Ket qua duoc thd bifen trfen hmh 3.
Hinh 3. Su thay ddi ciia hdm lupng protein theo flidi gian thiiy phdn vd loai enzyme protease Tir ket qua phan tich d bmh 3 cho thay theo thdi gian thuy phan ham lupng protein hda tan tao thdnh trong tat ca cdc mdu thuy phan dk\i tang nhung miic dp tang ciing khdc nhau tiiy thupc vao loai enzyme protease sir dung trong thi nghiem. Trong dd, hdm luong protein boa tan cua mau thuy phan hdn hpp sun ca map hdng hdn hpp enzyme alcalase + papain cd miic do tang nhanh theo thdi gian thiiy phdn va tang cao hon so vdi cdc mdu tiiuy phan khdc. Cu Ui^, d giai doan sau 2 gid thiiy phan, ham linmg protein cua mau thuy phan sun cd map bdng hdn hpp enzyme alcalase + papam cao gap 2,6 ldn so vdi ban ddu vd cao gap hrong ling 1,67 lan, 1,92 Idn, 2,02 ldn vd 2,19 ldn so vdi ham lupng protein boa tan t90
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thanh d mdu thuy phan bdng enzyme papain, alcalase, flavourzyme va neutrase. O thdi di^m sau 10 gid thiiy phan, ham lupng protein cua mdu thuy phan sun cd map bdng hdn hpp enzyme alcalase + papain cao gap tdi 5,6 ldn so vdi ban dau vd cao gap tuong ling 1,67 lan, 2,39 lan, 2,89 lan va 3,58 lan so vdi ham lirpng protein boa tan tao thdnh d mdu thuy phan bang enzyme papam, alcalase, flavourzyme va neutrase. Ket qua nay cho thay hdn hpp enzyme alcalase + papain cd kha nang thuy phan hdn hop sun ca map manh me hon so vdi cac enzyme papain, alcalase, flavourzyme va neutrase. Ket qua con cho thdy papain va alcalase khi boat ddng mpt minh ciing cd kha nang thuy phan hdn hop sun ca map tdt hon cac enzyme flavourzjmie va neutrase.
Kdt qua nay co the ly giai: cau tnic md sun ndi chung va sun cd map noi ri^ng, bSn ngodi dupe bao bdng cdc protein mo hen k^t, khong tan nhu elastin vd ben trong Id cdc protein khdng tan chiia cdc acid amin mach ben co cue vd ky nude [5, 7]. Trong khi dd elastin la protein chi bi thuy phan bdi papain. Do vay, khi su diing papain phdi hpp vdi alcalase thi papain se thuy phan protem md h6n kdt tao dieu kien cho alcalase phan cat cac protem ua nude d phia trong giai phdng ra dudi dang hda tan nen ham lupng protem hoa tan tao thdnh trong dung dich tang theo thdi gian thiiy phan.
Tir nhiing phan tich b trdn cho thay ndu xet theo khia canh ham lupng protem hda tan thi su diing hdn hpp enzyme alcalase + papain dd thuy phan sun cd map cd uu the la tao ra ham lupng protein hda tan h-ong dung dich tang nhanh theo thdi gian thuy phan vd tang cao hon cdc enzyme protease khac da su dung.
3.2. Anh hudng ciia thdi gian thuy phdn vd loai enzyme protease tdi hdm lugng peptid hda tan
Hinh 4. Su thay ddi ciia hdm lupng peptid thep thdi gian thiiy phan vd l o ^ enzyme protease
Tien hdnh 5 mdu thi nghidm thuy phdn hdn hpp sun ca map bdng 5 loai enzyme: neutrase, flavourzyme, alcalase, papam va hdn hpp enzym alcalase - papam va sau cac khoang thdi gian: 0 gid, 2 gid, 4 gid, 6 gid, 8 gid va 10 gid, tien hdnh lay mdu dich thuy phan de danh gia ham luong peptid hda tan Ket qud dupe the hien tren hinh 4.
Tir ket qua phan tich d hinh 4 cho thay ciing gidng nhu su thay ddi ciia hdm lupng protem trong qua hinh thuy phan, theo thdi gian thiiy phan ham lupng peptid tao thanh trong tat ca cdc mdu thiiy phan deu tang va miic dp tang cung khac nhau tiiy thupc loai enzyme protease. Trong do, hdm lupng peptid ciia mdu thuy phan hon hpp sun cd map bdng hdn hpp enzyme alcalase + papain tang nhanh hon theo thdi gian thuy phan va tang cao ban so vdi cdc mdu thuy phan khdc. Cu the, sau 2 gid thuy phan, ham lupng peptid cua mdu thuy phan su dung hdn hpp enzyme alcalase + papain cao gap 2,39 idn so vdi ban dau vd cao gap tuong ling 1,52 lan, 1,64 lan, 1,86 ldn vd 2,09 lan so vdi hdm lupng peptid tao thanh d mdu thiiy phan bdng enzyme papain, alcalase, flavourzyme vd neutrase. Tirong tu nhu vay, sau 10 gid thuy phan, ham lupng peptid ciia mdu thiiy phan sun ca map bang bdn hpp enzyme alcalase + papam cao gap tdi 2,96 lan so vdi ban ddu vd cao gap tuong ling 1,5 lan, 1,66 lan, 1,93 ldn va 1,99 ldn so vdi ham lupng peptid tao thanh d mdu thuy phan sun cd map bdng enzyme papain, alcalase, flavourzyme va neutrase. Ket qua ndy cho thay hdn hpp enzyme alcalase + papain c6 khd nang thuy phan protein cua sun cd map manh me hon so vdi cdc enzyme papain, alcalase, flavourzyme vd neutrase. Kdt qua cdn cho thay papain vd alcalase cting cd kha nang thuy phan protein ciia sun cd map tot bon cdc enzyme flavourzyme vd neutrase.
Tu nhiing phdn tich d tren neu xet theo khia canh ham lupng pephd tao thdnh thi su dung hdn hpp enzyme alcalase + papain de thuy phan sun ca map se tao ra thanh ham lupng peptid trong dung dich tang manh theo thdi gian thiiy phdn va tang cao hon cac enzyme protease khdc da su dung.
3.3. Anh huong ciia thdi gian thiiy phan vd loai enzyme protease tdi ham higng Naa
Tien hdnh 5 man thi nghiem thtiy phan hdn hpp sun ca map bdng 5 loai enzyme: neutrase, flavourzyme, alcalase, papain va hdn hpp enzym alcalase - papam vd sau cdc khoang thdi gian; 0 gid, 2
KHOA HOC CbNO NGHi gid, 4 gid, 6 gid, 8 gid va 10 gid, tidn hdnh lay dich
thuy phan de ddnh gid ham lupng Naa. Ket qua dupe the hien h-en hinh 5.
flavourzyme, alcalase, papam vd hdn alcalase - papain va sau cdc khodng thm •' gid, 4 gid, 6 gid, 8 gid vd 10 gid, ti^n n.-^
thiiy phan de danh gia hdm lupng sulphate.. K^t qua dupe the hi§n fren hinh
enzym ,.ji 0gid,2 idi lay dich
chodroitiii 6.
Hinh 5. Sy tiiay ddi cua hdm lugng Naa theo thdi gian thuy phSn vd l o ^ enzyme protease Ttr ket qua phan tich d hinh 5 cho thdy ciing gidng nhu su thay doi cua ham luong protein va peptid trong qua trinh thuy phan, theo thdi gian thuy phan ham lupng Naa tao thanh trong tat cd cdc mdu thuy phdn d^u tang vd miic dp tang cung khac nhau tuy thuOc loiu enzyme protease. Ham lupng acid amin tb^ hien qua chi sd hdm lupng Naa cua mdu thuy phan bang hdn hpp enzyme alcalase + papain tang nhanh theo thdi gian thuy phan va tang cao hon nhieu so vdi cac mdu thuy phan sun ca map bdng cdc enzyme protease khdc. Sau 10 gid thuy phdn, ham lupng Naa cua mau thuy phan sun cd map bang hdn hpp enzyme alcalase + papain cao gdp tdi 7,26 ldn so vdi ban ddu vd cao gap tuong ling 2,0 ldn, 2,11 lan, 2,61 ldn vd 3,06 lan so vdi ham lupng Naa tao thanh d mdu thuy phan sun ca map bdng enzyme papain, alcalase, flavourzyme va neutrase. Ket qua nay cho thdy hdn hop enzyme alcalase + papain cd kha nang thuy phan protein cua sun cd map thdnh acid amin manh me hon so vdi cdc enzyme papain, alcalase, flavourzyme vd neuh-ase. Ket qua ciing cho thay papam vd alcalase cung co kha nang thuy phan protein ciia sun ca map thdnh acid amine tot hon cac enzyme flavourzyme vd neutrase.
Tii nhiing phan tich d h-en ndu xet theo khia canh ham luong acid amm tao thanh thi su dung hdn hpp enzyme alcalase + papain de thiiy phan sun cd map se tao ra thdnh hdm lupng acid amin trong dung dich cao hon cdc enzyme protease khdc da sii dung.
3.4. Anh hudng ciia thdi gian thuy phan vd loai enzyme protease tdi hdm lupng chodroitin su^hate t^o thdnh
Tien hdnh 5 mdu thi nghidm thiiy phan hdn hpp sun cd map bdng 5 Ipai enzyme: neutrase.
Hinh 6. S^r thay ddi dia hdm lugng chondroitin theo thdi gian tiiiiy philn vd lo^i enzyme protease Tii ket qua phan tich d hinh 6 cho thay ciing gidng nhu d trgn, theo thdi gian thuy phan ham lupng chodroitin sulphate tao thdnh d tat ca cdc mdu thuy phan deu tdng nhanh va miie dd tang eiing khac nhau tiiy thupc loai enzyme protease. Trong do, hdm lupng chodroitin sulphate cua mdu thuy phdn sun ca map bang hdn hpp enzyme alcalase + papam tao thanh tang nhanh theo thdi gian thuy phan vd tang cao hon so vdi cdc mdu thuy phan bdng cac enzyme protease khac. Cu thd, sau 10 gid thiiy phan, hdm lupng chondroihn sulphate ciia mdu thuy phan s p cd map bdng hdn hpp enzyme alcalase + papain cao gap tdi 74,45 ldn so vdi ban dau va cao gap tuong ling 1,83 lan, 1,89 lan, 3,55 lan vd 4,92 lan so voi ham lupng chondrotin sulphate tao thdnh d mdu thiiy phan sun cd map bdng enzyme papain, alcalase.
flavourzyme va neutrase. Kdt qua nay mpt lan nia cho thay bdn hpp enzyme alcalase + papain co kha nang thuy phan sun cd map giai phdng chondroitin sulphate manh me hon so vdi cdc enzyme papain, alcalase, flavourzyme vd neutrase.
Tii nhirng phan tich d tren neu xet theo khia canh ham lupng chondroitin sulphate tao thanh thi sii dung hdn hpp enzyme alcalase + papain d^ thiiy phan sun ca map se tao ra thanh ham litp^ig chondroitm sulphate cao hon cac enzyme protease khac da su dung.
3.5. Anh hirdng cua tiidi gian thiiy phdn vd lo?i enzyme protease tdi ham hipng N^HS
Tidn hdnh 5 mdu thi nghiem thiiy phan hdn hcp sun cd map bdng 5 loai enzyme: neuhase, flavourzyme, alcalase, papam va hdn hpp enzym alcalase - papam vd sau cac khoang tiidi gian: 0 gicr, - 100
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gid, 4 gid, 6 gid, 8 gid vd 10 gid, tidn hanh lay dich tiiuy phan d^ ddnh gia hdm lupng NNH3 Ket qua dupe the hi^n trdn hmh 7.
Hmh 7. Sy: thay ddi ciia hdm lugng NNHS tiieo thdi gian thiiy phan vd lo^i enzyme protease Tir kdt qua phan tich d hinh 7 cho thay ham lupng NNH3 tao thdnh trong cdc mau thuy phan theo thdi gian d^u tang nhung miic dO tang chdm vd khac nhau khdng nhi^u. Cu th^, sau 10 gid thuy phan, cac mdu thuy phdn sun cd map bang enzyme: neutrase, alcalase, flavourzyme, papain va hdn hpp enzym alcalase - papam cd ham lupng NNH3 cao tuong ling so vdi ban ddu Id 1,3 lan, 1,27 ldn, 1,32 lan, 1,29 lan vd 1,27 ldn so vdi ham lupng N^HS ban ddu. Ket qua nay cho thdy cdc mdu thdy phan sun cd map bdng hdn hpp enzyme alcalase + papain, enzyme alcalase va papam cd ham lupng N^HS t^o tiianh thap ban chiit it so mdi cdc mdu thiiy phan khdc nhtmg miic dp chenh lech khdng dang ke.
4.K^UUiNViiKBttNGHI
- Cdc enzyme: neutrase, alcalase, flavourzyme, papain vd hdn hpp enzym alcalase - papain deu cd khd nang thuy phdn sun ca map d didu ki^n pH tu nhi^n, ty 1^ enzyme bd sung 0,2%, lupng nude bd sung 100% va nhiet do tiiuy phan 50*'C.
- Trong Cling dieu ki^n thuy phdn d pH tu nhien ciia hon hpp sun ca map, lupng nude bd sung 100%, ty le enzyme bd sung 0 ^ vd nhi^t dp thiiy phan 50"C, hon hpp enzym alcalase - papain cd kha nang thuy phan sun ca map tao thanh lupng protem hda tan, peptid, acid amin, chodroitin cao hon nhi^u lan so vdi cac enzyme khac da thu nghidm.
TAILJ£U THAM KHAO
1. Pham Thi Hai Au, Nguyen Tliuy Hudng (2010).
Nghiin cuu ung dung cong nghi enzym dS sin xuit sin phim protein thuy phin tirca tap viphSUiu tiong nhi miy chi' bii'n ca. Bdo cdo khoa hpc, Vidn Cdng
nghiep Thue pham Ha Ndi.
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KHOA HOC C b N G NGHfe
EFFECTS OF HYDROLVnC TIME AND PROTEOLYnC ENZYME TO THE HYDROLYHC PROCESS OF SHARK CARTHAGE {Carcharhinus dussumien)BY PRCfTEASE
Dmh Huu Dong, Vu Ngoc Boi, Nguyen Tin My Trang Summary
In this paper, we present a part of research results on hydrolyzing shark cartilage {Circharhmus dussumieri) by protease enzyme. Our results showed that the enzymes: neutrase, alcalase. flavourzyme, papam and the mixture of enzyme alcalase - papain all have the ability to hydrolyze shark cartilage mixture at natural pH, the enzyme supplement rate of 0.2%, 100% additional water and SO^C hydrolysis temperature.
Under the above conditions, the enzyme alcalase - papain enzyme is able to hydrolyze shark cartilage to create much higher soluble protein, peptide, ammo acid, chodroitin sulphate than hydrolyzed shark cartilage with other protease enzymes.
Keywords: Neutrase, alcalase, flavourzyme, papain, chodroitin sulphate, shark cartilage, hydrolysis.
Ngudi phdn bien: TS. Trdn Tlii Mai Ngdy nh&n bdi: 18/02/2020 Ngdy tiidng qua phan bi^n: 18/3/2020 Ngdy duyet dang: 25/3/2020
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