SỬ DỤNG SÓNG SIÊU ÂN TRÍCH LY polysaccha

Su dung song siéu :im trich ly polysaccharide LV M€J DAU Hon mfit nua dim so trén thé giai trfing va sit dung city thuong thuc nhu la ngufin luong thuc chu dao nhu: cay lua, cay lua mi, dai mach Nhiing thong tin khoa hpc gan day da nghién corn va phat hien lao gifing cay lvong thtic la chia khoa viec phat trién thuc ph:inn (Mutisya v:i cong su 2009) Polysaccharide va lignin duos xem la nhiing phan chinh co c:iy lupng thiic dupc xem la nhfiri g phan chinh va quan trong viec car thién sirc khoe (Yang va cfing su 2008a) nhu: day manh su trao dfii chat, xoa d|u than kinh, loai bo chfmg m:it ngu (Yang v:i cong sir 2008b; 2009) Polysaccharide trich ly tit nguon dong vat va thuc vat dune biét nhu I:i ngufin phu gia tu nhién cho nhiéu nganh cfing nghiép, dam biét la thuc pham va dune pham (Forabosco va cfing su 2006) Polysaccharide gin vat trfi quan trong sinh truong va phat trién cua thé b:to v:i dune nghién elm nhiéu nhiing nam g:in dfiy co nhfrng dupe tinh sinh hoc (Schepetkin va Quinn 2906) Rat it bao cao khoa hoc thuc ph:im vé trich ly polysaccharide co c:iy luong thuc, mfit nhfmg ly lii thiéu cfing nghé v:i phuong phap, phuong phap trich ly truyén thong bang hoi nude, bang dung moi thuéng khfing mang lai hieu qua cao, ton chi phi nang lvpng trich ly o nhiét cao, thai gian dai hieu suat thiip (Li v:i cfing su 2007) Do nhfrng han ché do, co mot nhu cau cho mfit phuong phap tot hon ma hiéu qu:i dé pha vo cac lién két mfi t thai gian ngan v:i thu dune nhiéu polysaccharide Nding luong sie‘ti rim da cho thfiy mot xu htifing trién vpng de‘ dat dupc muc tie‘ii Song siéu rim la ung dung cua cac song ém tan sfi v:i cuéng dfi cao (khoang 20— 100 kHz) de truyén Qua ch:it long ho:ie Tac dong chu yéu cua song siéu ém la truyén va tuong tac lain thay dfii tinh chat vat ly va hoa hoc cua vat liéu tae dung xiim thuc Xam thuc gay cue bfi o nhiét dfi cao v:i ap luc cao, két qu:i tao nhiéu gfic tu do, chang han nhu OH-, H ‘, va H;O;, do tang cufing cite ph:in ring hoa hpc Hieti ting cua sie‘u rim gin p thing cuong sts xam nhap ctia cac dting moi va nhiet vao té bao nguyén liéu do cai thién tot kha uang truyén khoi Song siéu am ciing co tac dung sir pha vo cac té bao sinh hpc de tao thuan loi cho viec giai phong dich can trich ly Nhom Trang I Su dung song siéu :im trich Iy polysaccharide 1.1 Polysaccharide v:i frng dpng Polysaccharide la cac gluxit phuc v‹ii phs tit r:it 1‹in gfim nhiéu don vi monosaccharide lién két v‹ii tao nén Polysaccharide khong co v| ngot nhu monosaccharide hay disaccharide, khfing tan nuoc ma chi tao dung d]ch keo Day la nhom chart him cc bién va co khoi luorig lan nhat tre‘n trai diit Polysaccharid rat da dang vé chung loai Trong cc the smh vfit co rat nhie‘u loai polysaccharide khac nhau, bién nh:it Ta tinh bot, glycogen, cellulose 1.1.1 Tinh b jot Tinh bot la ch:it du trii r:it phfi bien ‹ thuc vat Co nhiéu cac nhu hat, cu Tinh bot khfing phai la don chat ma 1:i hon hop cac chuoi thang cac phan tu amylose va chuoi phan nhanh la amilopectin Ty lé nhom chat tinh bot qii yét dinh cac tinh chfit ly - hoa cua chimg, quyét dinh chat lti9ng cua chung (do deo, nd ) * A invlose Amylose l:i polysaccharide dune tao nén ter cac M o.D.glucose Cac o.D glucose lién két voi bang lién két (I o - phan 4) glucozid tao nén chii oi polysaccharide Moi lién két glucozit ducrc tao se‘ loai mot phan tu HCO Do chi co loai lien ke't (In - 4) glticozid chin tao ne‘n amylose ne‘n phan tu amylose co cau trite mach thang Hinh : Mach amylose Nhom Trang Su dung song siéu :im trich ly polysaccharide Amylose dune tao tu 5000 - 1000 phan tu o.D glucose (co chi khoang 250 - 300 phan tu) Chuoi phan tu glucose xoan lai voi theo hinh xoan 1o xo St hinh dang xoan hinh thGh cac lién két hydro giua cac glucose tao Moi vong xoan co don vi glucose va dvpc dtiy tri boi lién ket hydro vcri cac vong xoan ké bén Khoang khfing gian gifra cac xoan co kich thuoc phu hpp cho mot khac ién két v:to, vi du nhii iod Khi phan ttt iod ién két vao vfin g xo an se lam cho cac phan tu glucose thay dfii vi tri chut it v:i tao nén phuc in:iu xanh dam trung Dang xoan cua amylose chi tao dung dich c nhiét thuéng Khi c nhiet dfi cao chuoi xo¼ se bi duoi thug va khfing co kha nang lie‘n két voi cac phs tu khac * Am vlopectin Amylopectin co cau tao phiic tap hon Tham gia cau tao amylopectin co khoang 500.000 dén I trieu phan tu o-D-glucose lién két voi Trong amylopectin co loai lién két: — Lién két o (I — 4) glucozid tao mach thang - Lién két o (1- 6) glucozid tao mach nhanh Hinh 2: Mach amyl opectin Chi khoang 24 - 30 don vi glucose trén mach lie‘n két o (1 - 6) glticozid de‘ tao mach nhanh Trén mach nhanh cap lai hmh mach nhanh cap 2, cv nhv vay Su dung song siéu :im trich ly polysaccharide phan tfr amylopectin ph:in nhanh nhiéu c:ip r:it phuc tap.Trong tinh bot ty lé amylopectin chiém khoang 80%, amylose chiém 20% Ty lé n:iy thay doi o cac nhom sinh vat khac nhau.Tinh bfit I:i nguyén liéu du trii thuc viit Day la dang du trn thich hpp nhat vi tinh bot khfing co kha nang thfim qua mang Ie bao nén khfing thé that thoat khoi té bao 1.1.2 Glycogen Glycogen lii polysaccharide du trfr o dong vat, la tinh bfit ‹ dong vat Cau trtic cua glycogen giong tinh bfit nhirng muc ph:in nhanh nhiéu hon fi tinh bot, cu khoang - 12 don vi glucose da co mot lién két o (I — 6) glucozid dé tao nh:inh mii O dong vat v:i nguoi, glucogen dune du trii chin yéu o gan Su phan huy v:i tong hop glycogen dune thong cac hoocmon diéu khién mot cach chat ché dé diéu hon su on dinh lvpng glucose mau luon la so I% 1.1.3 Cellulose Trong cac hop chat hiiu cc co cc thé smh vat thi cellulose co ty lé cao hon ca No lii phs chinh cua té b:to thuc vat Ciing nhu amylose, amylopectin, cellulose la chat triing hop tu nhiéu don ph:in Thanh ph:in don phan cua cellulo se la Q-D-glucose Cac ph:in ter Q-D-glucose lién két vfii bang lién két Q (I — 4) glucozid thay "siip" v:i I "ngua" Su thay dfii vé th:inh phiin v:i ciiu tao dan dén su khac biét vé tinh chat giiia cellulose va amylose Phan tfr cellulose khfing cufin xoan nhu amylose ma chi co c:iu tribe dang mach thang Cau trim tao diéu kién hinh cac lién két hydro gifra cac phan tu cellulose nam song song vfii nhau, tao nén cau trtic inning cellulose va vi soi (micro fibrin) ciiu trim m:ing cellulose cua té bao thuc v:it Cac soi khfing tan nude, rat bén vé cc hoc nén tao nén lip mang cellulose bén chac Hinh :Cau trtic lién két Q (I — 4) glucozid 1.2 Song siéu am 1.2.1 D{nh nghia Siéu am (ultrasound) la song cp hpc hinh su lan truyén dao dong cua cac phan tu khong gian co tfin so tin hon gicri han trén nguéng nghe cua ngucri (16-20kHz) Ngoai ra, song siéu ann co b:in chat lii song doc (longitudinal wave) hay song nén (compression wave), nghia I:i trufing siéu am cac phan M dao dong theo phuong ciing voi phtiong tniyén ciia song Utrasound lnfr asound Upper-ranQng8als Bumble bee Middle U Human hearing Mos9uitu Power ultrasounds Crasshop9er Extended range High frequency Hinh 4: Phfin loai cac loai song theo tan so Su dung song siéu :im trich Iy polysaccharide 1.2.2 Born chfi t cfia séng fi m bén moi truéiig nay, no vi tri can bring be‘n Do tuong tac tao nén boi cac moi lién két vc i cac ph:in tir bén canh, mot m:it phs tit A b| kéo vé v| tri c:in bang, mot meat no ciing chin tae dung boi luc tac dong nén phan M A se di chuyén qua — lai quanh vi tri can bang, co nghia la ph:in tu A thuc hien chuyén dong duoi dang dao dong Hien tuong tiép tuc xay doi vat cac phan tfr khac cua moi truéng Dang dao dong cc, co tinh ch:it lap di lap lai, lan truyén moi tru‹ing dan hoi dune goi I:i song d:in hfii hay song cc, noi mot cach khac, song la mfit hién tuong vat ly nding luong dune an ttrruuyéénn duoi dang dao dong cua cac phan tu vat chat cua moi truérig truyén song Ve ban chat, song rim la song cc hoc, do no turn theo mpi quy liiat doi vcri song cc, co the tao song am bang cach tac dong mot Inc cc hpc vao moi truong truyén am Vd 1: Tac dong mot luc lam ning le‘n fim thoa, gfiy cho cac phfin ki khong bi nén lai hay dan lily theo hv rig chii ye‘n dong cua am thoa, phfin tu dfiu tién bi tac dong se anh huong dén phan tit ké tiép v:i cu thé ma co su lan truyén song m pi hii6ng (va cling nho thé ma tai ngufii o bat ky v] tri nao xung quanh finn thoa déu nghe du9c am vang cua finn thoa) Hien tupng tuong tu nhu ta tha mot vién soi v:to gifra long hfi dang lang sing, vién soi sé tao nhfrng gun song co hinh dang cac vfing trfin dong tain lan toa xung quanh ma Um cua chung la vi tri ma vién soi roi xtiong hfi nufic Vd 2: Danh vao mat trfi ng; tac dong dong dien lam rung mang loa; dan bay khong 1.2.3 Cite dai lu irng dac trirng ciia si'i ng Su dung song siéu :im trich ly polysaccharide Hinh bén duc i la hinh biéu dién cua song, no la mfi t tap hop cua cac l:in nén va dan thay dfii tu:in tit theo dang hinh sin, cac dinh song the hien ap luc cao nhat cac duty song thé hién ap luc thap nhat Hinh 5: Do thi biéu dié'nn ddaaoo dong cua song am Cite dai luong dam trung cua song bao gfi m: • Chu ky T = (s) la khoang thai gian ma song thuc hién mot lan nén va mfit lan dan • Tan so I= (Hz) la so chu ky thuc hien dupe giay • Van toc truyén cua song finn la qtiang dvong ma song rim tniyén dupc sau mot don vi thai gian • Do dai buoc song \ = (gin): la quang duéng ma song truyén duoc sau khoang thcri gian bang I chii ky (\ = v.T = v/f) Tre‘n hinh ve‘, ta thay btioc song \ la khoang cach gifra hai dinh hoac hai day nam ke' 1.2.4 Phén loai sling fim Phil n loai theo phirirng dao dong: dna vao cach truye‘n song, ngtifii ta chia song cc lam hai loai: song doc va song ngang Su dung song siéu :im trich Iy polysaccharide • Song ngang la song ma phuong dao dong cua cac ph:in tfr cua moi truéng vuong goc voi tia song Song ngang xufit hien cac moi truéng co tinh dan hfii vé hinh dang Tinh chat n:iy chi co o vat ran • Song doc lit song in:i phuong dao dong cua cac phiin tu moi trufing trting vfii tia song Song d pc xuiit hien ca moi truéng chin bién dang vé the tich, do no truyén duos cac vat ran ciing nhii moi trying long v:i Song siéu am ting dung sie‘ii am chfin doan thuoc loai song dpc Phil n lo9i theo tan so: song ann dvpc chia theo dai tin so vfing chinh • Song :im t:in ' cuc th:ip, hay got la song am (Infrasound): I 16 Hz Vi du: song dia chan • Song am tG so nghe thay dune (Audible sound): I= 16 Hz — 20 kHz • Song siéu am (Ultrasound): I > 20kHz Cac ngufin song siéu rim co tu nhién: Doi, mfit viii lo:ii ca bién phat song siéu am dé dinh hu6ng Koi chung cac song n:inn vfing t:in so 20 — 00 kHz Song sie‘ti rim kg ding y hoc co tG so tu 700 KHz den 50 MHz siéu am chan doan su dung cite tan so tit MHz dén 50 MHz 1.2.5 Ph9in vi frng dung Siéu ann la mot linh vuc dang diioc nghién cfiu va co tiém n:ing phat trién nganh cong nghé thtic phs Song siéu finn co tin so tit 20kHz den trén 25 MHz thuéng duoc ring dung rong rai nhiéu linh c Co linh vuc dupe ring dung chinh cfing nghiép thuc pham: T:in so cao va nang lupng th:ip: siéu ém chuan doan, khoang tan so MHz Phan duoc sts dung nhu mot ky thuat phan tich dam bao chat luong, qui trinh diéu khién v:i kiém tra khfing him pha huy ciiu trfic, diéu nity dune frng dung xac dinh tinh chat thuc pham, toe dfi dong chiiy, kiém tra bao goi thuc phfim .( Floros & Liang, 1994; McClements, l995 ; Mason, Paniwnyk & Lorimer, 1996; Mason 1998) Su dung song siéu :im trich ly polysaccharide T:in so thap va siéu :im nang luong cao: Phan duoc ting dung rong rat nhu mot guy trinh ho tro h:ing 1oat cac tinh c nhu: két tinh, s:iy, bai khi, trich ly, loc, dong hoa, l:inn mém thit, qua trinh oxi hoa, qua trinh tiét triing (Floros & Liang, I 994;Gennano et al.,1999; Mason, l 998; Mason et al., 1996; McClements, l 995) Being : Mfii vai ung dung cua song siéu am cfing nghé thuc pham Eng dpng c:ic hieu frng c‹ hpc ting dpng hieu frng hfia hpc vii hfia sinh Qua trinh két tinh ch:it béo, duéng Diét khu:in Bai Xu ly dong ch:iy (nude thai) Pha bpt Diéu chinh ski sinh truén g ciia te' bao Trich ly cite chat thom song Ho tro lpc va say tach Bien doi hoat tinh enzyme him Lanh dong Tiét triing thiét bi cfing nghiép Khuay trfin v:i dong hoa Két cac hat bui In limg Lam mém thit 1.2.5.1 Béo quén thu’c phfim Vfi ho9t vi sinh vat Cfing nghiép ché bién thuc ph:inn thu‹ rig tap tiung chu yéu vao qua trinh vo hoat (inactivation) hoac tie‘u diet cac vi sinh vat va enzyme nham muc dich ke’o dai thcri gian biro quiin s:in phiim Rat nhiéu phuong phap vat ly dune ap dung v:to muc dich nay, phuong phap dung nhiet cao la bién va hieu qua tiéu diet vi sinh vat cao nhat The nhimg vi c su dung nhiet cao co the gay mot so bién doi bat loi doi vcri san pham nhu lam thiit thoat mo I so chfit dinh during va lam giam cac tinh ch:it cam quan cua thuc ph am Do do, yéu c:iu d(at cho cac nh:i nghién elm k thuat thuc pham la tim cac giai phap mii, cac phuong phap v:it ly khac dé thay thé ho:ie ho tro mo I phan phuong phap nhiét Qua trinh trying va tiét trying san ph:inn Su dung song siéu :im trich Iy polysaccharide Qua trinh siéu am két hop vat ap su:it (manosonication-MS) dune thuc hién bfii mot so tac gia nhu Sat a v:i cong su (1995), Man as va cong sir (2000) trén cac doi tuong vi sinh viit gay bénh ciing cho thay nhfrng két qua riit kha quan Cite nghién Thu cho thfiy tai nhie‘t dfi thuéiig qua trinh xv ly sie‘u ém khong high qiia vfii A monocytogenes (véi thoi gian chét nhiét la D - 4.3 phut) Bang phuong phap siéu am két hop voi ap suat (MS), gia tri D cua vi khu:in giam xufi ng l phut Khi gia nhiét dén 50’C, hiéu qu:i cua qua trinh xu ly khfing dang ké nhung o nhiét dfi cao hon thi hieu qua xfr ly lai tfi t hon r:i t nhiéu Mfi t diém dang luu y dfii voi kj thu at siéu ann so voi kj thuat triing nhiet truyén thong la c phuong phap nhiét, kha nang va cc ché dé khang cua vi khuan la yéu to quan can duoc quan tain qua trinh xfr ly Nhimg v‹ii ky thuat MTS va MS, kha nang dé khang nhiét cua vi khuan hfiu nhu khfi ng co Do do, ky thuat mang lai nhiéu trién vong hon bfii hiéu qua xu ly cua no khfing phu thufic veto diéu kién xfi ly va kha nang chfing chiu nhiet cua vi khuan thuc pham to vfii Qua trinh nhiet Bén canh do, nghie‘n cfru cua Pagan va cong su (I 999) cho thay rang ski ton thuong cua té b:to boi nhiét dfi la thu an nghich nhirn g ton thupng g:iy boi qua trinh xu ly MTS l:i bat thuan nghich Phuong phap su dung song siéu am cu rig cao la mot phuong phap hieu qua viec khiiay tron cac phan tu nho, tang kha nang truyén khfii Co che' va phvon g phap tac dong cua song siéu am dupc Penn va cfing su (1959) giai thich rang song siéu am va su ho tro cua may khuay nho sé tao bot gan bé mat cua chat ran co thé giam lip bién truyén khfii va do lam tang su truyén khfi i Co ché qua trinh tao nhii tuong: Khi tae dung song siéu :im vao ch:it long, su tao lo trfi ng xuat hién ap suat cua song dat dén mot gia tri xac dinh to i thiéu n:to do, gia tri nity dune goi I:i nguéiig tao lo trfing Trong thong dau/nuns, qua trinh nhfi hoa xay dat den mfrc nguérig Song sie‘ti rim co the ciing cap mot nding liiong du cho viec hinh th:inh bé in(at mii, vi thé no co thé lain cho qua trinh nhii hoa xay ca khong dung chat nhfi hoa Néu bong bong b| vo g:in 16p bién cua Su dung song siéu :im trich Iy polysaccharide bang may nghién bua Theo cac nha cung c:ip,qua ph:in tich HPLC phan xylan nguyén chat lfii trung binh I:i 32,6% Thiét bj siéu am va cirfrng sling am Bo hat dau dfi siéu :im: a Model 53000 (Misonix Inc, NY) vfii cfing sufit 350 W, tin so song am dao dong tronn khoang 20 kHz, b Model IL 1000-6/2 (cfing ty TNHH Saint-Petersburg, Nga), lam viec o tan so 22 kHz,cfi ng suat I OOO W Mat dfi nding luong song am (AED) dune khiio sat cho ca hai miiy siéu rim su dung nghién cfru Cuéng siéti am cua may phu thuoc vao khoi luong va the tich man lfiy phan tich Doi vcri cac mati phfin tich, nhiet (T) du9c ghi lai bang mot nhiet ké xem la mo I ham cua thoi gian Ty Ie thing nhiet (AT/At) dupc tinh khoang 30s/1:in Mat n:ing luong song am cho moi may diioc tinh bang cong suat trung binh chia cho thé tich nufic sit dung thti nghiém Két qua Mat dfi nang luong song rim cho cii hai may dune tom tat bring Xfr 1v/ s‹r bo ltii bii p bang phuong phap thfi ng thirfrng Mau Hi bap 10g dune ngam axit sulfuric lo iing (HMSO 2%, ty lé : 10 khfii Ivpng / the tich) fi 511 'C 24 gig (hinh l ) Chat trich ly da du9c tach bang Ipc Luong acid dv luong du9c ma sach bang nvoc tiucrc hap Xu ly so bo pha vfi cac lién két hoa tri gifra lignin va xylan loi bap, va giai phong xylan Su dung song siéu :im trich ly polysaccharide Hinh l6: So sanh phuong phap: xii ly thong thuéng va xii ly str dung song siéu ann Ngam lfii bap dung dich HMSO 2% tai 10,20,30 va 60 phut vfii mat dfi song ann lan luot 0.18W Xii l/v sir bo liii bép bang sling siéu fim Trich ly Can l0g Hi bap rig:inn axit sulfuric loang (HMSO, 2%, ty lé I : 10 khfii luong / thé tich) viing chac dé ty lé ch:it long) sau chiéu song siéu am 10, 20, 30 hoax 60 phut o mat dfi nding luong song am la 0,45 W / mL va 0, 18 W / ml Cac mau dem lpc va rtia sach bring voi nucrc truoc http (Hinh 16) Trich ly Trich ly Mau dune dat mot ong big thép khfing gi dem h:ip riot hap H:ip 30 phfit tai nhiét dfi 10, 118, hoax 121 ’C (Hinh 16) (Yang vii cfing su.) va nhimg khoang nhiét duoc xem trich ly toi da xylan va giam thiéu viéc giam duéng va cac san pham khfing mong mufin khac San hap, mati tron vfii 100 ml nufic o 16009 rad/ phiit phut Doi vfii phfin tich hoa hoc, chfit long thu dvpc sau trich ly dem loc Cac man l pc dem phfin tich hoa hoc su hien dién cua xylan Phil n tich hfia hpc cfin c:i c chit trich ly Xylan thu dune dau cac qua trinh trén dem di xac dinh big cach tong so duéng hoa tan Theo Yang va cong giai thich rang: tong so dufing hoa tan chart Nhom Trang 34 Su dung song siéu :im trich ly polysaccharide chiét bao gfim xylan hoa tan va xylooligosaccharides, nhung luong xylooligosaccharides la rfit thap v:i cellulose hoa tan lfii bap I:i kh fing dang ké quit trinh xu ly so bfi va trich ly, tong so hfia tan duéng l:i mfit phuong phap tvong doi chinh xac dinh lvpng xylan mati Mat khac, xylooligosaccharides ciing san phfim thuan loi cua qua trinh trich ly, boi vi sau enzyme chuyén dfii xylan, cac san pham chu yéu la xylooligosaccharides Do do, thu duos duoc xylan hoax xylooligo saccharides nhu qua trinh trich ly Boi vi diéu nay, nghién erm so luong nho cac xylooligosaccharides duoc tinh vao dich trich ly dé don gian xac dinh xylan Do do, luong xylan duoc tinh b:ing cach ham luong duéng tong so hfia tan nuns loc, nhu thé hién duoi day : Ltiong xylan dich trich ly Luong xylan co dich trich(% ' Luong xylan nguyén lieu ‘ 3.2.3 Két quit vii thao lu a( n Trich ly xylan sau xu ly so bfi (truyén thong) 24 h va hap 30 phfit, lap thi nghiém nhu trén ‹i cac nhiét dfi khac (I 10, 118, va 12 I 'C) nhu bang Khoang, 29,1 % xylan d:in tién bap duoc trich ly (Bang 2) Http mang lai hieu qua trich ly nhiéu xylan hon vi dp: ty Ie‘ trich ly du9c 7,3%, 8,2% va 8,8% tai 10, 118, va l2l 'C Diéu cho thay http nang cao trich ly xylan, co thé vi hai ly sau day: - Nhiéu lién két cong hoa tri cua lignin va carbohydrate khac b] pha huy qua trinh http, do lam tang kha nang trich ly — Mac dii lfii bap dune rira sach bang nucc tru‹ic hap nhung van cfin co mot so HMSO sot lai vii o nhiét dfi cao acid nity thuy phan ’ co thé diéu ra.Tuy nhién, truce dfiy phupng phap co the da dong mot vai tro lon trich ly xylan 110 B:ing 3: Cac thiing phap trich ly truyén thfi ng Thong ké Turkey chi rang ty lé trich ly http tai 118 and 121 'C khac dang ké vat ty le trich ly tai 10 'C (p