L-thioglutamic acidyog ib qho khoom siv ntuj tsim uas tau hloov pauv los ntawm glucosinolate zoo li precursors hauv grannymonda zaub xws li kale, mustard zaub thiab zaub paj. Nws muaj ntau yam teebmeem kev noj qab haus huv, thiab tau dav siv los tiv thaiv cov kab mob ntev xws li mob qog noj ntshav, kab mob plawv, thiab ntshav qab zib. Nws yog dav pom nyob rau hauv cov nroj tsuag cruciferous, xws li kale, cauliflower, thiab cabbage. Nws muaj ntau yam tshuaj ua haujlwm, xws li tshuaj tiv thaiv kab mob, tshuaj tiv thaiv kab mob, tshuaj tua kab mob, tshuaj tiv thaiv kab mob, thiab lwm yam. Yog li ntawd, cov txheej txheem hluavtaws ntawm L-thioglutamic acid tau nyiam ntau yam.
Tam sim no, cov txheej txheem rau kev sib txuas L-thioglutamic acid tuaj yeem muab faib ua ob pawg: lom fermentation thiab tshuaj synthesis. Ob txoj kev no tau piav qhia hauv qab no.
L-Sulforaphane biological fermentation txoj kev:
Cov txheej txheem biological fermentation yog ib txoj kev ntawm kev sib txuas nrog cov kab mob. Los ntawm kev tshuaj xyuas cov kab mob thiab kev ua kom zoo ntawm kab lis kev cai, cov kab mob tuaj yeem tsim L-thioglutamic acid thaum lub sijhawm fermentation. Tam sim no nws tau pom tias ntau hom tuaj yeem tsim cov L-thioglutamic acid, xws li Escherichia coli, Streptococcus thermophilus thiab lwm yam. Ntawm lawv, txoj kev rau kev sib txuas L-thioglutamic acid los ntawm cov kab mob uas sawv cev los ntawm Escherichia coli yog feem ntau. Cov kauj ruam tshwj xeeb yog raws li hauv qab no:
1. cais L-thioglutamate-producing hom, xws li Escherichia coli, los ntawm cov av los yog cov ntaub so ntswg;
2. Txais cov kev cai nruab nrab thiab kab lis kev cai, xws li kub, pH nqi, kab lis kev cai lub sij hawm, thiab lwm yam., kom loj hlob Escherichia coli;
3. Sau cov Escherichia coli fermentation broth, nqa tawm tej yam kev ua thiab purification, thiab tau L-thioglutamic acid.
L-Sulforaphane chemical synthesis method:
Cov txheej txheem tshuaj synthesis yog siv cov txheej txheem ntawm cov tshuaj molecular cov tshuaj tiv thaiv los ua ke L-thioglutamic acid los ntawm kev sib txawv hluavtaws. Tam sim no, cov txheej txheem tshuaj synthesis feem ntau muab faib ua plaub hom: choline ntsev txoj kev, sulfuryl chloride txoj kev, carbonyl txo txoj kev, thiab agarose sulfate txoj kev.
1. Choline ntsev txoj kev:
Txoj kev no yog ib txoj hauv kev uas siv hluavtaws. Ua raws li hauv qab no:
① Ua ntej, L-methionine thiab choline reacted nyob rau hauv acidic tej yam kev mob los tsim ib tug tsawg nitrogen hloov ntawm L-thioglutamic acid.
② Cua sov thiab hydrolyzing nitrogen hloov rau hauv L-thioglutamic acid nyob rau hauv alkaline tej yam kev mob.
Qhov kom zoo dua ntawm txoj kev no yog tias cov tshuaj tiv thaiv yog yooj yim thiab yooj yim rau tus tswv, tab sis cov tawm los yog tsawg.
2. Txoj kev sulfonyl chloride:
Txoj kev no siv cov tshuaj electrophilic hloov pauv los ua ke L-thioglutamic acid. Ua raws li hauv qab no:
① Cov tshuaj tiv thaiv L-methionine thiab sulfonyl chloride nyob rau hauv alkaline tej yam kev mob kom tau L-thioglutamic acid sulfinate.
② Cov sulfinate yog hydrolyzed nyob rau hauv alkaline tej yam kev mob los tsim L-thioglutamic acid.
Qhov kom zoo dua ntawm txoj kev no yog tias cov tshuaj tiv thaiv tau yooj yim los tswj thiab cov txiaj ntsig tau siab, tab sis nws yuav tsum tau siv tshuaj lom sulfonyl chloride.
3. Carbonyl txo txoj kev:
Cov txheej txheem yog siv txo cov tshuaj tiv thaiv los ua ke L-thioglutamic acid. Ua raws li hauv qab no:
①L-methionine reacts nrog acetonimine los tsim pyridine pyrimidin-2-en-4-ib thiab pyrimidine pyrimidin-2-en-4-ib-5-sulfate tib lub sijhawm
②Pyrimidine pyrimidine-2-en-4-ib -5-sulfate txo kom tau L-thioglutamic acid.
4. Agarose sulfate txoj kev: Txoj kev no yog ua kom L-methionine thiab agarose sulfate nyob rau hauv acidic tej yam kev mob los tsim L-thioglutamic acid. Vim qhov zoo catalytic kev ua si ntawm sulfate, cov tshuaj tiv thaiv efficiency thiab selectivity yog txhim kho. Siab dua, tab sis ntau cumbersome rau kev khiav hauj lwm.
Hauv kev tshawb fawb tshawb fawb, kev tshawb pom ntawm L-sulforaphane tseem ceeb heev. Ntawm lawv, txoj kev agarose sulfate yog ib qho ntawm cov kev tshawb nrhiav classic. Txoj kev agarose sulfate thiab nws cov kauj ruam yuav piav qhia hauv qab no.
Kev sim txoj cai
Txoj kev agarose sulfate yog tsim los ntawm kev tsim cov xim daj ntawm L-thiogalactopicrin thiab barium sulfate, ntsev tsim nyob rau hauv acidic tej yam kev mob, siv agarose raws li ib tug scaffold cais L- Txoj kev sulforaphane. Cov txheej txheem yog raws li qhov tseeb tias nyob rau hauv acidic tej yam kev mob, lub daj complex tsim los ntawm cov tshuaj tiv thaiv ntawm barium sulfate thiab L-sulforaphane tsis yooj yim soluble nyob rau hauv dej thiab precipitates nyob rau hauv agarose, thiaj li mus cais thiab ntes L-sulforaphane.
Kev sim cov kauj ruam:
Kauj Ruam 1: Npaj lub agarose-barium sulfate ncua kev kawm ntawv
Ntsuas tus nqi tsim nyog ntawm agarose thiab barium sulfate, ntxiv ib qho dej distilled tom qab sib tov thiab do kom huv si, cua sov kom txog thaum yaj los yog sib tov sib xyaw, lim lub ncua kev kawm ntawv nrog lub lim nrog lub pore loj ntawm 0.22um, thiab ces boil rau 3 mus rau 5 feeb. Thaum qhov kev ncua txias mus rau qis dua 50 degree tab sis tseem tsis khov, ncuav cov tshuaj rau hauv Petri tais thiab muab tso rau hauv chav tsev kub kom txog thaum nws solidifies.
Kauj Ruam 2: Qauv npaj thiab ua tiav:
Ntsuas tus qauv lossis tus qauv yuav tsum tau kuaj, ntxiv kom txaus {{0}}.01 M sodium pyrophosphate tov kom yaj, ntxiv cov ntim ntim ntawm chloroform rau rho tawm, centrifuge rau 10-15 feeb , tshem tawm cov txheej txheej sab saud, evaporate cov chloroform kom khaws cov seem, ntxiv 2 mL dej thiab 2 mL ntawm concentrated hydrochloric acid, muab rho tawm dua nrog chloroform, centrifuged, thiab cov txheej sab saud raug coj mus. Siv 24mL ntawm cov kua dej sab saud, ntxiv 0.2 mL ntawm 2 feem pua tshuaj tooj liab nitrate thiab 0.4 mL ntawm 5 feem pua ntawm cov tshuaj nitrate, rho tawm ib zaug nrog chloroform, centrifuge, thiab nqa cov kua sab saud, uas tuaj yeem siv los ua cov qauv kev sim rau kev sim.
Kauj Ruam 3: Kev Tshawb Fawb Txog Kev Tshawb Fawb:
Cov qauv tau ntxiv rau agarose-barium sulfate ncua kev kawm ntawv thiab muab tso rau hauv ib da dej kom khov ntawm 80 degree rau 2 teev. Tom qab cov qauv yog solidified, lub agarose-barium sulfate ncua kev kawm ntawv yog txias rau hauv chav tsev kub, thiab cov agarose-barium sulfate-cov qauv complex yog yaj nyob rau hauv tsis muaj tej yam kev mob ntawm citric acid thiab ammonium ntsev. Thaum kawg, yaug nrog dej ua ntu zus, thiab pom cov xim ntawm lub complex nyob rau hauv lub agarose sphere.
Kauj Ruam 4: Kev Tshawb Fawb Txog Kev Tshawb Fawb:
Ntsuas qhov nqus ntawm cov lim dej tom qab reanalysis los tsim cov qauv nkhaus, yog li tau txais cov ntsiab lus ntawm L-sulforaphane.
Agarose sulfate txoj kev yog ib txoj kev classic rau kev sib cais thiab kuaj L-sulforaphane. Los ntawm ntau cov kauj ruam, L-sulforaphane tuaj yeem sib cais thiab kuaj pom nws cov ntsiab lus. Cov txheej txheem muaj qhov zoo ntawm kev yooj yim, siab rhiab heev, thiab cov txiaj ntsig raug, thiab feem ntau siv los kuaj xyuas L-sulforaphane hauv cov khoom noj, tshuaj pleev ib ce, thiab tshuaj.
Cov ntsiab lus: Nrog kev txhim kho ntawm kev tshawb fawb thiab thev naus laus zis, cov txheej txheem ntawm kev sib txuas L-thioglutamic acid tau ua kom zoo thiab txhim kho tsis tu ncua. Nyob rau tib lub sijhawm, txoj kev siv tshuaj lom neeg fermentation thiab cov tshuaj sib xyaw ua ke kuj muaj lawv tus kheej qhov zoo thiab qhov tsis zoo, thiab txoj kev siv twg yog nyob ntawm qhov xwm txheej tiag tiag.