Lub tswvyim ntawm immobilized enzymes thawj zaug tshwm sim nyob rau hauv ib nrab ntawm lub xyoo pua 20th. Lub caij no, rov qab rau xyoo 1916, nws tau pom tias sucrose sorbed ntawm carbon khaws nws cov haujlwm catalytic. Xyoo 1953, D. Schleit thiab N. Grubhofer tau ua thawj qhov kev sib txuas ntawm pepsin, amylase, carboxypeptidase thiab RNase nrog rau cov cab kuj insoluble. Lub tswv yim ntawm immobilized enzymes tau raug cai hauv xyoo 1971. Qhov no tshwm sim ntawm thawj lub rooj sib tham ntawm engineering enzymology. Tam sim no, lub tswv yim ntawm immobilized enzymes tau txiav txim siab nyob rau hauv ib qho kev dav dav dua li qhov kawg ntawm lub xyoo pua 20th. Cia peb saib ze rau ntawm pawg no.
cov ntaub ntawv dav dav
Immobilized enzymes yog cov khoom sib xyaw ua ke uas ua rau muaj kev cuam tshuam rau cov khoom thauj tsis zoo. Txawm li cas los xij, lawv khaws lawv cov khoom catalytic. Tam sim no, cov txheej txheem no tau txiav txim siab nyob rau hauv ob yam - nyob rau hauv lub moj khaum ntawm ib feem thiab ua tiav kev txwv ntawm kev ywj pheej ntawm kev txav ntawm cov protein molecules.
Nyob zoo ib tsoom niam txiv kwv tij neej tsas sawv daws.
Cov kws tshawb fawb tau tsim qee cov txiaj ntsig ntawm immobilized enzymes. Ua raws li heterogeneous catalysts, lawv tuaj yeem yooj yim sib cais los ntawm cov tshuaj tiv thaiv nruab nrab. Raws li ib feem ntawm kev tshawb fawb, nws tau pom tias kev siv cov immobilized enzymes tuaj yeem rov ua dua. Thaum lub sij hawm txoj kev khi, kev sib txuas hloov pauv lawv cov khoom. Lawv tau txais substrate specification thiab stability. Nyob rau tib lub sijhawm, lawv cov haujlwm pib nyob ntawm ib puag ncig ib puag ncig. Immobilized enzymes yog ruaj thiab muaj kev ruaj ntseg siab. Nws yog ntau dua, piv txwv li, ntawm cov enzymes dawb los ntawm ntau txhiab, kaum tawm txhiab zaus. Tag nrho cov no ua kom muaj kev ua haujlwm siab, kev sib tw thiab kev lag luam ntawm cov thev naus laus zis uas muaj cov enzymes immobilized tam sim no.
Media
J. Poratu tau txheeb xyuas cov khoom tseem ceeb ntawm cov ntaub ntawv zoo tshaj plaws los siv hauv immobilization. Bearers yuav tsum muaj:
- Insolubility.
- High lom lom thiab tshuaj tiv thaiv.
- Lub peev xwm ua kom sai sai. Cov neeg nqa khoom yuav tsum yooj yim ua reactive.
- Qhov tseem ceeb hydrophilicity.
- Nyob rau permeability. Nws qhov taw qhia yuav tsum muaj qhov sib npaug ntawm cov enzymes thiab coenzymes, cov khoom siv tshuaj tiv thaiv thiab cov substrates.
Tam sim no tsis muaj cov khoom siv uas ua tau raws li cov cai no. Txawm li cas los xij, hauv kev xyaum, cov cab kuj tau siv uas tsim nyog rau kev tsis tuaj yeem. Qee qeb ntawm enzymes nyob rau hauv tej yam kev mob.
Kev faib tawm
Nyob ntawm lawv qhov xwm txheej, cov ntaub ntawv, nyob rau hauv kev sib txuas nrog cov tebchaw uas hloov mus rau hauv immobilized enzymes, muab faib ua inorganic thiab organic. Kev khi ntawm ntau lub tebchaw yog nqa tawm nrog cov khoom siv polymeric. Cov ntaub ntawv organic no tau muab faib ua 2 chav kawm: hluavtaws thiab ntuj. Nyob rau hauv txhua tus ntawm lawv, nyob rau hauv lem, pawg yog txawv nyob ntawm seb tus qauv. Inorganic carriers feem ntau yog sawv cev los ntawm cov khoom siv iav, ceramics, av nplaum, silica gel, thiab graphite dub. Thaum ua hauj lwm nrog cov ntaub ntawv, qhuav chemistry txoj kev yog nrov. Immobilized enzymes tau txais los ntawm cov txheej txheem nrog cov yeeb yaj kiab ntawm titanium, aluminium, zirconium, hafnium oxides lossis los ntawm kev ua haujlwm nrog cov organic polymers. Ib qho txiaj ntsig tseem ceeb ntawm cov ntaub ntawv yog qhov yooj yim ntawm kev tsim dua tshiab.
YProtein carriers
Cov khoom nrov tshaj plaws yog lipid, polysaccharide thiab cov khoom siv protein. Ntawm cov tom kawg, nws tsim nyog hais txog cov qauv txheej txheem polymers. Cov no feem ntau suav nrog collagen, fibrin, keratin, thiab gelatin. Xws li cov proteins tau nthuav dav hauv ib puag ncig ntuj. Lawv pheej yig thiab pheej yig. Tsis tas li ntawd, lawv muaj ntau pawg ua haujlwm rau kev khi. Proteins yog biodegradable. Qhov no tso cai rau nthuav dav siv immobilized enzymes hauv tshuaj. Lub caij no, cov proteins kuj muaj cov khoom tsis zoo. Qhov tsis zoo ntawm kev siv cov enzymes immobilized ntawm cov neeg nqa khoom muaj protein ntau yog qhov kev tiv thaiv kab mob siab rau yav tom ntej, nrog raulub peev xwm los qhia txog qee pawg ntawm lawv rau hauv kev tawm tsam.
Y Polysaccharides, aminosaccharides
Ntawm cov ntaub ntawv no, chitin, dextran, cellulose, agarose thiab lawv cov derivatives feem ntau yog siv. Txhawm rau ua kom cov polysaccharides tiv taus ntau dua rau cov tshuaj tiv thaiv, lawv cov kab sib txuas yog sib txuas nrog epichlorohydrin. Ntau pawg ionogenic tau qhia pub dawb rau hauv cov qauv hauv network. Chitin accumulates nyob rau hauv ntau ntau raws li pov tseg thaum lub sij hawm ua lag luam ntawm cws thiab crabs. Cov tshuaj no yog tshuaj resistant thiab muaj ib tug zoo-txhais tus qauv ntxeem tau.
YSynthetic polymers
Cov ntaub ntawv no muaj ntau haiv neeg thiab siv tau. Nws suav nrog cov polymers raws li acrylic acid, styrene, polyvinyl cawv, polyurethane thiab polyamide polymers. Feem ntau ntawm lawv yog mechanically muaj zog. Nyob rau hauv tus txheej txheem ntawm kev hloov pauv, lawv muab qhov muaj peev xwm ntawm qhov sib txawv ntawm qhov pore loj nyob rau hauv ib qho dav dav, qhia txog ntau pawg ua haujlwm.
Kev khi txoj kev
Tam sim no, muaj ob txoj hauv kev sib txawv rau kev immobilization. Thawj yog kom tau cov khoom sib txuas yam tsis muaj covalent bonds nrog cov cab kuj. Txoj kev no yog lub cev. Lwm qhov kev xaiv suav nrog qhov tshwm sim ntawm kev sib cog lus nrog cov khoom siv. Nov yog chemical method.
Adsorption
Nrog kev pab los ntawm nws, immobilized enzymes tau txais los ntawm kev tuav cov tshuaj nyob rau saum npoo ntawm tus neeg nqa khoom vimdispersion, hydrophobic, electrostatic kev sib cuam tshuam thiab hydrogen bonds. Adsorption yog thawj txoj hauv kev los txwv kev txav ntawm cov khoom. Txawm li cas los xij, txawm tias tam sim no qhov kev xaiv no tsis tau poob nws qhov tseeb. Ntxiv mus, adsorption yog suav tias yog txoj kev immobilization feem ntau hauv kev lag luam.
Txoj kev ntawm txoj kev
Cov ntawv tshaj tawm tshawb fawb piav qhia ntau dua 70 enzymes tau los ntawm txoj kev adsorption. Cov cab kuj yog cov iav ntxeem tau, ntau yam av nplaum, polysaccharides, aluminium oxides, hluavtaws polymers, titanium thiab lwm yam hlau. Cov tom kawg yog cov feem ntau siv. Qhov ua tau zoo ntawm adsorption ntawm cov tshuaj ntawm tus neeg nqa khoom yog txiav txim siab los ntawm qhov porosity ntawm cov khoom thiab thaj chaw tshwj xeeb.
Mechanism of action
Enzyme adsorption ntawm cov ntaub ntawv insoluble yog yooj yim. Nws yog ua tiav los ntawm kev sib cuag ntawm aqueous tov ntawm cov tshuaj nrog cov cab kuj. Nws tuaj yeem dhau ntawm txoj kev zoo li qub lossis dynamic. Cov tshuaj enzyme yog tov nrog cov sediment tshiab, piv txwv li, titanium hydroxide. Lub compound yog ces qhuav nyob rau hauv me me tej yam kev mob. Enzyme kev ua haujlwm thaum lub sijhawm immobilization yog khaws cia los ntawm yuav luag 100%. Nyob rau tib lub sijhawm, qhov tshwj xeeb concentration nce mus txog 64 mg ib gram ntawm cov cab kuj.
Neegative moments
Qhov tsis zoo ntawm adsorption suav nrog lub zog qis thaum khi cov enzyme thiab cov cab kuj. Nyob rau hauv tus txheej txheem ntawm kev hloov cov tshuaj tiv thaiv tej yam kev mob, poob ntawm cov ntsiab lus, paug ntawm cov khoom, thiab protein desorption yuav raug sau tseg. Txhawm rau txhim kho lub zogkhi cov cab kuj tau hloov kho ua ntej. Tshwj xeeb, cov ntaub ntawv raug kho nrog hlau ions, polymers, hydrophobic compounds, thiab lwm yam polyfunctional agents. Qee zaum, cov tshuaj nws tus kheej raug hloov kho. Tab sis feem ntau qhov no ua rau txo qis hauv nws txoj haujlwm.
Inclusion nyob rau hauv lub gel
qhov kev xaiv no muaj ntau heev vim nws qhov tshwj xeeb thiab kev yooj yim. Txoj kev no yog tsim tsis tsuas yog rau ib tug neeg lub ntsiab, tab sis kuj rau multi-enzyme complexes. Kev koom ua ke rau hauv cov gel tuaj yeem ua tau ob txoj hauv kev. Nyob rau hauv thawj rooj plaub, cov tshuaj yog ua ke nrog ib tug aqueous tov ntawm lub monomer, tom qab uas polymerization yog ua. Raws li qhov tshwm sim, cov qauv spatial gel tshwm, muaj cov enzyme molecules hauv cov hlwb. Nyob rau hauv rooj plaub thib ob, cov tshuaj tau nkag mus rau hauv cov kev daws teeb meem ntawm cov polymer tiav. Nws muab tso rau hauv lub xeev gel.
Kev nkag mus rau hauv cov qauv translucent
Lub ntsiab lus ntawm txoj kev immobilization no yog kev sib cais ntawm cov kua dej enzyme los ntawm substrate. Rau qhov no, ib tug semi-permeable membrane yog siv. Nws tso cai rau cov khoom hnyav molecular tsawg ntawm cov cofactors thiab substrates kom dhau mus thiab khaws cov khoom loj ntawm cov enzymes.
YMicroencapsulation
Muaj ntau ntau txoj kev xaiv rau embedding nyob rau hauv cov qauv translucent. Ntawm cov no, microencapsulation thiab kev koom ua ke ntawm cov proteins rau hauv liposomes yog qhov txaus siab tshaj plaws. Thawj qhov kev xaiv tau npaj rau xyoo 1964 los ntawm T. Chang. Nws muaj nyob rau hauv qhov tseeb hais tias cov tshuaj enzyme yog nkag mus rau hauv ib tug kaw capsule, cov phab ntsa uas yog ua los ntawm semi-permeable.polymer. Cov tsos ntawm ib daim nyias nyias ntawm qhov chaw yog tshwm sim los ntawm cov tshuaj tiv thaiv ntawm interfacial polycondensation ntawm cov tebchaw. Ib tug ntawm lawv yog yaj nyob rau hauv cov organic, thiab lwm yam - nyob rau hauv lub aqueous theem. Ib qho piv txwv yog tsim ntawm microcapsule tau los ntawm polycondensation ntawm sebacic acid halide (organic theem) thiab hexamethylenediamine-1, 6 (feem ntau, aqueous theem). Lub thickness ntawm daim nyias nyias yog xam nyob rau hauv pua pua ntawm ib micrometer. Qhov loj ntawm cov tshuaj ntsiav yog ntau pua lossis kaum tawm micrometers.
Kev koom ua ke rau hauv liposomes
Txoj kev immobilization no yog ze rau microencapsulation. Liposomes muaj nyob rau hauv lamellar los yog kheej kheej systems ntawm lipid bilayers. Txoj kev no tau siv thawj zaug hauv xyoo 1970. Txhawm rau cais cov liposomes los ntawm cov tshuaj lipid, cov kuab tshuaj organic yog evaporated. Cov yeeb yaj kiab nyias nyias tau tawg mus rau hauv cov kua dej uas muaj cov enzymes tam sim no. Thaum lub sijhawm ua haujlwm no, kev sib sau ua tus kheej ntawm cov txheej txheem lipid bilayer tshwm sim. Xws li immobilized enzymes yog nrov heev hauv cov tshuaj. Qhov no yog vim lub fact tias feem ntau ntawm cov molecules yog nyob rau hauv lub lipid matrix ntawm biological membranes. Cov immobilized enzymes suav nrog hauv liposomes yog cov ntaub ntawv tshawb fawb tseem ceeb tshaj plaws hauv cov tshuaj, uas ua rau nws muaj peev xwm kawm thiab piav qhia cov qauv ntawm cov txheej txheem tseem ceeb.
Kev tsim cov ntawv cog lus tshiab
Immobilization los ntawm kev tsim cov kab sib txuas tshiab ntawm cov enzymes thiab cov nqa khoom yog suav tias yog txoj kev dav tshaj plaws rau kev tau txais cov khoom siv biocatalysts.qhov chaw. Tsis zoo li cov txheej txheem ntawm lub cev, qhov kev xaiv no muab ib qho irreversible thiab muaj zog daim ntawv cog lus ntawm molecule thiab cov khoom. Nws tsim yog feem ntau nrog cov tshuaj stabilization. Nyob rau tib lub sijhawm, qhov chaw ntawm cov enzyme nyob ntawm qhov deb ntawm 1st covalent daim ntawv cog lus txheeb ze rau tus neeg nqa khoom tsim muaj qee qhov teeb meem hauv kev ua raws li cov txheej txheem catalytic. Cov molecule raug cais tawm ntawm cov khoom siv los ntawm kev ntxig. Nws feem ntau yog siv los ua poly- thiab bifunctional agents. Tshwj xeeb, lawv yog hydrazine, cyanogen bromide, glutaric dialhedride, sulfuryl chloride, thiab lwm yam. Piv txwv li, kom tshem tawm galactosyltransferase, cov kab lus hauv qab no tau muab tso rau ntawm cov cab kuj thiab cov enzyme -CH 2 - NH-(CH2)5-CO-. Nyob rau hauv cov xwm txheej zoo li no, ib qho ntxig, ib qho molecule, thiab cov cab kuj muaj nyob hauv cov qauv. Tag nrho cov ntawm lawv yog txuas nrog covalent bonds. Qhov tseem ceeb ntawm qhov tseem ceeb yog qhov yuav tsum tau ua kom nkag mus rau hauv cov tshuaj tiv thaiv kev ua haujlwm uas tsis yog qhov tseem ceeb rau kev ua haujlwm catalytic ntawm lub caij. Yog li, raws li txoj cai, glycoproteins txuas nrog rau cov cab kuj tsis yog los ntawm cov protein, tab sis los ntawm cov carbohydrate ib feem. Yog li ntawd, cov enzymes ruaj khov thiab nquag immobilized tau txais.
Cells
Cov txheej txheem tau piav qhia saum toj no suav tias yog universal rau txhua hom biocatalysts. Cov no suav nrog, ntawm lwm yam, cov hlwb, cov txheej txheem subcellular, immobilization ntawm uas nyuam qhuav dhau los ua dav. Qhov no yog vim cov hauv qab no. Thaum cov hlwb raug immobilized, tsis tas yuav cais thiab ntxuav cov enzyme npaj los yog qhia cov cofactors rau hauv cov tshuaj tiv thaiv. Yog li ntawd, nws yuav ua taucov txheej txheem uas ua tiav ntau theem txuas ntxiv.
Kev siv cov enzymes immobilized
Hauv cov kws kho tsiaj, kev lag luam, thiab lwm yam kev lag luam, cov tshuaj tau los ntawm cov txheej txheem saum toj no yog nrov heev. Cov txheej txheem tsim tawm hauv kev xyaum muab kev daws teeb meem rau cov teeb meem ntawm kev xa tshuaj hauv lub cev. Immobilized enzymes ua rau nws muaj peev xwm tau txais cov tshuaj ntawm kev ua haujlwm ntev nrog kev ua xua thiab toxicity tsawg. Tam sim no, cov kws tshawb fawb tab tom daws cov teeb meem cuam tshuam nrog bioconversion ntawm huab hwm coj thiab lub zog siv cov txheej txheem microbiological. Lub caij no, thev naus laus zis ntawm immobilized enzymes kuj ua rau muaj txiaj ntsig zoo rau kev ua haujlwm. Kev cia siab rau txoj kev loj hlob zoo li dav heev. Yog li ntawd, nyob rau hauv lub neej yav tom ntej, ib tug ntawm cov tseem ceeb luag hauj lwm nyob rau hauv tus txheej txheem ntawm kev soj ntsuam lub xeev ntawm ib puag ncig yuav tsum tau mus rau cov tshiab hom kev tsom xam. Hauv particular, peb tab tom tham txog bioluminescent thiab enzyme immunoassay txoj kev. Cov txheej txheem Advanced yog qhov tseem ceeb tshwj xeeb hauv kev ua cov khoom siv raw li lignocellulosic. Immobilized enzymes tuaj yeem siv los ua cov teeb liab tsis muaj zog. Lub chaw nquag tuaj yeem raug cuam tshuam los ntawm tus neeg nqa khoom uas nyob rau hauv ultrasound, kev nyuaj siab, lossis raug hloov pauv ntawm phytochemical.
