Amidase (AMD)
MaEnzayimu:Mapuroteni ndiwo mapuroteni anokonzera mamorekuru, ma enzyme mazhinji ndiwo mapuroteni
Midase:Kuvandudza hydrolysis yemarudzi akasiyana-siyana e endogenous neangaphandle aliphatic ne aromatic amides nekutamisa boka re acyl mumvura nekugadzirwa kwe free acids ne ammonia. Hydroxamic acids nedzimwe organic acids dzinoshandiswa zvakanyanya semishonga nekuti inhengo dze growth factors, antibiotics uye tumor inhibitors. Ma amidases anogona kukamurwa kuita R type ne S type acylases zvichienderana ne catalyst stereoselectivity.
Pamusoro pekukonzera hydrolysis yeamides, amidase inogonawo kukonzera acyl transfer reactions kana paine co-substrates dzakadai se hydroxylamine.
MaAmidase ane manyuko akasiyana ane ma substrate specificity akasiyana, mamwe acho anogona chete ku hydrolyze amides aromatic, mamwe anogona chete ku hydrolyze aliphatic amides, uye mamwe ano hydrolyze α-kana ω-amino amides. Mazhinji emaamine ane catalytic activity yakanaka chete kune acyclic kana simple aromatic amides, asi kune complex aromatics, heterocyclic amides, kunyanya ma amides ane ortho substituents, anowanzo kuve mashoma mukushanda (ma enzyme mashoma chete anoratidza zviri nani catalytic effects).
Maitiro ekugadzirisa:
| MaEnzayimu | Kodhi Yechigadzirwa | Kodhi Yechigadzirwa |
| Upfu hweEnzimu | ES-AMD-101~ ES-AMD-119 | seti ye19 amidases, 50 mg chimwe nechimwe 19 zvinhu * 50mg / chinhu, kana chimwe chiyero |
| Kiti Yekuongorora (SynKit) | ES-AMD-1900 | seti ye19 amidases, 1 mg chimwe nechimwe zvinhu 19 * 1mg / chinhu |
★ Hunhu hwepamusoro hwe substrate.
★ Kusarudza kwakasimba kwerudo rwechikara.
★ Kushanda zvakanyanya pakushandura.
★ Zvibereko zvishoma.
★ Mamiriro ekuita zvinhu zvisina kusimba.
★ Inochengetedza zvakatipoteredza.
➢ Kuongororwa kwema enzyme kunofanirwa kuitwa kune mamwe ma substrate nekuda kwehunhu hwe substrate, uye kuwana enzyme inokatyamadza substrate yacho nemhedzisiro yakanaka yekukatyamadza.
➢ Usambosangana nemamiriro ezvinhu akaipisisa akadai se: tembiricha yakakwira, pH yakakwira/yakaderera uye organic solvent ine huwandu hwakawanda.
➢ Kazhinji, reaction system inofanira kunge iine substrate, buffer solution (Iyo yakanyanya kunaka pH ye enzyme). Co-substrates dzakadai se hydroxylamine dzinofanira kunge dziripo mu acyl transfer reaction system.
➢ AMD inofanira kuwedzerwa pakupedzisira mu reaction system ine pH uye tembiricha yakakwana.
➢ Mhando dzese dzeAMD dzine mamiriro akasiyana-siyana ekuita zvakanaka, saka imwe neimwe inofanira kudzidzwa imwe neimwe.
Muenzaniso 1(1):
Basa reHydrolysis kune akasiyana maAmide Substrates
| Substrate | Chiitiko chaicho μmols miniti-1mg-1 | Substrate | Chiitiko chaicho μmols miniti-1mg-1 |
| Acetamide | 3.8 | ο-OH benzamide | 1.4 |
| Propionamide | 3.9 | p-OH benzamide | 1.2 |
| Lactamide | 12.8 | ο-NH2benzamide | 1.0 |
| Butyramide | 11.9 | p-NH2benzamide | 0.8 |
| Isobutyramide | 26.2 | ο-Toluamide | 0.3 |
| Pentanamide | 22.0 | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | Nicotinamide | 1.7 |
| Cyclohexanamide | 19.5 | Isonicotinamide | 1.8 |
| Acrylamide | 10.2 | Picolinamide | 2.1 |
| Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Prolinamide | 3.4 | Indol-3-acetamide | 1.9 |
| Benzamide | 6.8 |
Mhinduro yacho yakaitwa mu 50mM sodium phosphate buffer solution, pH 7.5, pa 70 ℃.
| Madhimoni | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
Mhinduro yacho yakaitwa mu 50mM sodium phosphate buffer solution, pH 7.5, pa 70 ℃.
Kuwanda kwe reagent kwakabatana: amides, 100 mM(benzamide, 10 mM); hydroxylamine ne hydrazine, 400 mM; enzyme 0.9 μM.
Muenzaniso 2(2):
Muenzaniso 3(3):
1. D'Abusco AS, Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, nevamwe. Process Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, nevamwe. Bioorganic Chemistry, 2017, Inowanikwa online 7.
