BAO CAO T 6 M TAT " H O I N G H I K H O A HQC K * N | | M 40 N A M (1975-2015) j THANH LJBIP VigN HAN L A M KHOA HQC VA CdNG NGHf V l ^ NAM"
i^.^y> m^G c A C D l l U K l | N T 6 | U U A N H H U ' d m G T O I Q U A T R I N H Ti^iO ^ 0 D A I o ! l A P R O T E I N M Y O S I N T R O N G CO CA 0 ^ S A N X U A T C H A C A SJBLCH T C I I C A D O
c O PTEROCAESIO DIGRAMMA (BLEEKER, 1 8 6 4 ) ' - NguySn Thu Hong^, Nguyen Van D § ^ Nguyin Thanh Blnh^ Shugo Watabe'*
^Vi'in HaiDi/ang hoc, 01 Cdu Oa, Nha Trang, [email protected] 'casdsdn xudt Cha cd Hdng, 18 Cu Chl, Nha Trang, [email protected]
'OQihQC YHd N$l, 01 Tdn That Tdng, Ha NQI, [email protected]
^Bgl hQC Kftasato, NhgtBdn, [email protected]
Ttnh cdp thiit: Chd cd dugc xem Id nguon tiiuc phim khde bdi gidu protein vdi .3thft^.acid amin thiit ylu khdng thay till vd It cholesterol. Nd dugc sdn xuit tCr nhilu lodi cd^ibdc nhau pho biln Id cdc lodi cd gid tri kinh t l thip cd gid chg bdn rd. Neu nhiing d l i tirgng ndy dugc i ^ e n cihi d l chl biln thdnh nhilu sdn phim cd gid tri dinh dudng vd kinh t l cao nhu chd cd thi 5€ gdp phin eho vipc sd: dyng hieu qua vd b i n vftng nguon Igi sinh hpc biln vd ndng cao thu nh|ip cho ngu ddn (Shimizu vd cs., 1981; Matsuoka vd cs., 2013).
T ^ nude ta, chd cd dang is trong tinh hang nhilm bin do ngudi chl biln sii dyng .'DhOng hda chat vd ph\i gia dpc hai nhu hdn the, ur€ vd chloramphenicol de t ^ dp dai nSn rit cd h ^ eho siie khde cua ngudi tifiu diing. Tgi Phii Yen, Ding Thdp, 100% cdc ssff sd sdn xuit chd cd dugc kilm tra dd bi nhilm cdc hda chit trfin (Sdr Y t l Phli Y6n, 2013; Sdf Y t l Ding Thdp, 2013), tii|m chi tinh tr^ng sdn xuit chd cd bin cung da duge de c|p tren chuong trinh thdi sir trong cd nude vd dugc xem Id tinh trgng nhiic
• •aa^ci h i ^ nay (VTVl, 2013). Nguyen nhdn cOa tinh trgng ndy Id qui trinh chl biln cdn
"^uy OmOc vdo cac hda chat vd ph\i gia tgo dai bSn ngodi md chua cd nghiSn ctiu, ting 4\ing d l ndng cao Idia nang vd hieu qud ctia cdc protein tgo d$ kit dinh c6 sdn trong cd.
Protein tgo dai ed mgt nhilu trong c a cd, vdi tiidnh phin chii ylu Id cdc myosin va actin, cd tdc dyng tgo dp ddn h i i eua co dudi dnh hudng ciia nhipt dp (Yasui vd cs., i980). Cde myosin (MHC) trong th|t cd s€ c h i ^ ^ thdnh cdc myosin don phdn MHC(n), tgo cdc mgng ludi gel dudd tdc dung ciia nhiet dd vd cdc dieu k i ^ khdc trong qud trinh chl bien (Shimizu va cs 1981). Chinh vl vdy, dya tren nghien ciiu trudc ve dnh hudng ciia nhipt dd len qud trinh tao gel ciia jn'otein tgo dai myosin cua cd dd cii ( s l hpu chua cdng b l ) chiing ^ i d& xdc dinluibi|t dp toi uu cho qud trinh tgo gel dat cue dgi dao ddng trong khodog 39 - 41''C, do dd trong thi nghidm ndy chiing tdi dd thii n h i ^ d0 tii uu 41''C dudi dilu ki$n chuin nhu pH (7.0), hdm lupng m u l i Id 3% vd
B t o cAa TdM TAT - H O I N G H I K H O A Hgc KV Nr^M U N A M (1S75-201S) T H A N H liP V||N H A N L S M KHOA HOC VA C 6 N G NGHf VlfT NAM"
thdi gian iS (30 phiit) di t ^ gel len thit xay nhuyin cua cd dd Cd Pterocaesio digramma (Bleeker, 1864) (Id lodi thudng dupe sd dvmg di ldm chd cd t^i Nha Trang vd mgt s6 tinh, thdnh khdc) di iing dung sdn xuit chd cd s^ich, khdng hdn the vi phu
Phuai^phip nghiin ciu .
Xdc djnh su ddn hii cda gel td th}t cd dd Cd Pterocaesio digramma (Bleeker, 1864) xay nhuyai (tiieo Shimizu vd cs., 1981) tp diiu ki$n da tiiiit Idp.
Xdc djnh sv phdn buy cua MHC thanh cdc don phdn MHC(n) tiong gel cda thjt ca dd Cd Pterocaesio digramma (Bleeker, 1864) bdng SDS-PAGE (Webem and Obsom., 1969)
Ap dvmg qui trinh sdn xuat chd cd sach ra th\rc ti tuc diiu i d ^ t6i uutrSiL
Tim tit kit qud vi thao lufn
Sir dan hdi cia gel tii thft cd dd cii: Cd Pterocaesio digramma (Bleeker, 1864) xay nhuyen khi toi uu hda cdc diiu kien (pH: 7.0, 3% NaCl, nhi$t dg tao dd dai tii da trong budc u on djnh gel: 41°C, flidi gian xd 1^ nhift 30 phut trong qud trinh u nhiet vd 20 phut cho qud trinh hip chin d 85-90°C).
•i ,
s *
»**fltr<5
' • J
»Jld«««3
Hinh L B ; bin IN) id i)« dai |Kj ciia eel th|t d M ci xay nhuyin i ntu k l ^ tffi ini hiSa
Gel duijc in nhi^ d 41°C t?o d§ bin li 12,71 iO,25 N. Dd bin ndy cao hon 2 lin dii vdi mlu d6i Chung (khdng xdlynhi?t, nlm ti«n tryc tung)va 1,2 lan so vdi dd dai cue dai cua cd md (Nguyen Thu Hong vi cs., inpiess). So sdnh kit qud vi djic tinh gel cua cd dd cu vdi cdc cd khdc sing tp vdng dn ddi da du9c nghien ciiu nhu cd minh thdi (walley pollack), cd dd (white croaker) vd mpt sd loii cd xuong cho Ally dd bin vd dd dai cda gel ci dd gIp tir 4,2 - 6,2 lin trong trudng hpp khdng in dinh nhi6t vi gap tir 5,2-8,2 lin trong ttudng hpp dupe u ttong khoang nhiet d§ 40-45''C teudc khi hap chin (Matsuoka vd cs., 2013).
Srphan huy cia myosin thanh cdc dan phdn trong gel cua thit ca id cu: So vdi
BAO CAO T6M TAT 'HOt NGHI KHOA HQC KY N | | M 40 N&M (1975-2015) THANH L5J» VlfN HAN IAM KHOA HQC VA C 6 N 6 NGHf V I | T NAM"
nmu thjt xay nhuyen (SM) vd ddi chiing (Cont) khdng dagc xii 1^ nhiet trudc Idii h i p chin, vgch (band) MHC ciia mdu gel dugrc xii ly nhi^t d 41°C md hon rit nhilu, ngbia Id MHC da ehuyin tiidnh cdc don phdn MHC(n). Su liSn kit cua cdc MHC ndy dudi tdc dyng cua TGase tgo n€n d$ b i n vd d$ dai cua gel dupe do c^ till d k i t qua 1 (Hinh 1). Ket qud ndy p h i hgp vdi kit qua nghien ciiu cua Shimizu vd cs. (1981), Wan vd cs.
(1994), vd Matsuoka vd cs. (2013) tiiy theo m l i lodi cd, dr dilu ki$n tdc dOng t i i uu s6 d ^ den su phan hiiy MHC thdnh don phdn MHC(n), tang cirdng dO dai ciia sdn phim chd cd tur lodi cd do.
^ ^ H MHC(iiJ H
^ ^ B MHC- H
'1^1
Hinh 2. SDS-PAGE cda myosin sau khi dirgrc tiCl ini hda
Nhu v$y, vdi dilu ki$n pH: 7.0,3% NaCl, nhi|t do on djnh gel Id 41''C, flieri gian xii ly nhiSt: 30 phiit trong qud trinh u nhi?t va 20 phiit cho qud trinh h i p chin d 85- 90*'C tiii dO dai ciia gel cd dd cii dat dugc 12,7 N - dO dai t i i da cCia chd cd tii ed dd Cii Pterocaesio digramma (BleekCT, 1864).
Nhdm tdc gid da ^p dyng thanh cdng gid trinh sdn xudt chd cd tu cd do: Cii Pterocaesio digramma (Bleeker, 1864) tii dilu kifn da duge nghien ciiu tren tao nhilu loai sdn phim cha cd sach (100% Id cd nguySn chit), khdng hdn the, ph\i gia vd hda chdt md ddm bdo dg dai ddn theo thi h i ^ cua ngudi tiSu diing (phin tnmg b ^ sdn phim).
Tic khda: Cd do cd, chd cd sach, gel, myosin, SDS-PAGE, thit cd xay nhuyin
