I-Amidase (AMD)
Iienzymes:Zii-catalysts zebhayoloji ze-macromolecular, uninzi lwee-enzyme ziiproteni
I-Amidase:Yenza i-hydrolysis ye-aliphatic kunye ne-aromatic amides ezahlukeneyo ze-endogenous kunye nezangaphandle ngokudlulisela iqela le-acyl emanzini kunye nemveliso ye-free acids kunye ne-ammonia. Ii-Hydroxamic acids kunye nezinye ii-organic acids zisetyenziswa kakhulu njengeziyobisi kuba zizinto ezibangela ukukhula, ii-antibiotics kunye nezithinteli ze-tumor. Ii-amidases zinokwahlulwa zibe yi-R type kunye ne-S type acylases ngokwe-catalyst stereoselectivity.
Ukongeza ekuvuseleleni i-hydrolysis ye-amides, i-amidase inokubangela i-acyl transfer reactions xa kukho ii-co-substrates ezifana ne-hydroxylamine.
I-Amidase enemithombo eyahlukeneyo ine-substrate specificity eyahlukileyo, ezinye zazo zinokusebenzisa i-hydrolyze amides e-aromatic kuphela, ezinye zinokusebenzisa i-hydrolyze aliphatic amides kuphela, kwaye ezinye zinokusebenzisa i-hydrolyze α-okanye i-ω-amino amides. Uninzi lwee-amines zinomsebenzi omhle we-catalytic kuphela kwi-acyclic okanye i-simple aromatic amides, kodwa kwi-complex aromatics, i-heterocyclic amides, ngakumbi i-amides ene-ortho substituents, ngokubanzi ayisebenzi kakhulu (zimbalwa ii-enzymes ezibonisa iziphumo ezingcono ze-catalytic).
Indlela yokwenza i-catalytic:
| Iienzymes | Ikhowudi yeMveliso | Ikhowudi yeMveliso |
| Umgubo we-Enzyme | ES-AMD-101~ ES-AMD-119 | iseti ye-19 amidases, 50 mg nganye Izinto ezili-19 * 50mg / into, okanye olunye ubungakanani |
| Isixhobo sokuvavanya (iSynKit) | I-ES-AMD-1900 | iseti ye-19 amidases, 1 mg nganye Izinto ezili-19 * 1mg / into |
★ Ukucaciswa okuphezulu kwe-substrate.
★ Ukukhetha okunamandla kwe-chiral.
★ Ukusebenza kakuhle kokuguqula.
★ Iimveliso ezingaphantsi kwezinye.
★ Iimeko zokusabela okuncinci.
★ Inobuhlobo nokusingqongileyo.
➢ Ukuhlolwa kwee-enzyme kufuneka kwenziwe kwiindawo ezithile ngenxa yobungakanani be-substrate, kwaye kufunyanwe i-enzyme evuselela i-substrate ekujoliswe kuyo enefuthe elihle kakhulu le-catalytic.
➢ Ungaze udibane neemeko ezinzima ezifana nobushushu obuphezulu, i-pH ephezulu/ephantsi kunye nesinyibilikisi sendalo esinoxinzelelo oluphezulu.
➢ Ngokwesiqhelo, inkqubo yokusabela kufuneka iquke i-substrate, isisombululo se-buffer (I-pH yempendulo efanelekileyo ye-enzyme). Ii-co-substrates ezifana ne-hydroxylamine kufuneka zibekho kwinkqubo yokusabela kwe-acyl transfer.
➢ I-AMD kufuneka yongezwe okokugqibela kwinkqubo yokusabela ene-pH kunye nobushushu obufanelekileyo.
➢ Zonke iintlobo ze-AMD zineemeko ezahlukeneyo zokusabela ezifanelekileyo, ngoko ke nganye yazo kufuneka ifundwe ngakumbi nganye nganye.
Umzekelo 1(1):
Umsebenzi we-Hydrolysis kwii-substrates ezahlukeneyo ze-Amide
| I-substrate | Umsebenzi othile i-μmols min-1mg-1 | I-substrate | Umsebenzi othile i-μmols min-1mg-1 |
| I-Acetamide | 3.8 | ο-OH benzamide | 1.4 |
| I-Propionamide | 3.9 | p-OH benzamide | 1.2 |
| I-Lactamide | 12.8 | ο-NH2benzamide | 1.0 |
| I-Butyramide | 11.9 | p-NH2benzamide | 0.8 |
| I-Isobutyramide | 26.2 | ο-Toluamide | 0.3 |
| I-Pentanamide | 22.0 | p-Toluamide | 8.1 |
| I-Hexanamide | 6.4 | I-Nicotinamide | 1.7 |
| I-Cyclohexanamide | 19.5 | I-Isonicotinamide | 1.8 |
| I-Acrylamide | 10.2 | I-Picolinamide | 2.1 |
| I-Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| I-Prolinamide | 3.4 | I-Indol-3-acetamide | 1.9 |
| IBenzamide | 6.8 |
Isiphumo senziwe kwisisombululo se-50mM sodium phosphate buffer, pH 7.5, kwi-70 ℃.
| Amide | I-Hydroxylamine | IHydrazine |
| I-Acetamide | 8.4 | 1.4 |
| I-Propionamide | 18.4 | 3.0 |
| I-Isobutyramide | 25.0 | 22.7 |
| IBenzamide | 9.2 | 6.1 |
Isiphumo senziwe kwisisombululo se-50mM sodium phosphate buffer, pH 7.5, kwi-70 ℃.
Uxinaniso lwe-reagent oluhambelanayo: ii-amides, i-100 mM(benzamide, i-10 mM); i-hydroxylamine kunye ne-hydrazine, i-400 mM; i-enzyme eyi-0.9 μM.
Umzekelo 2(2):
Umzekelo 3(3):
1. U-D'Abusco AS, u-Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, et al. I-Process Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al. Bioorganic Chemistry, 2017, Ifumaneka kwi-intanethi 7.
