I-Amidase (AMD)
Ii-Enzymes:Ngaba i-macromolecular biological catalysts, uninzi lwee-enzymes ziiproteni
Amidase:Catalyze the hydrolysis of ezahlukeneyo endogenous kunye angaphandle aliphatic kunye amides aromatics ngokudlulisela iqela acyl emanzini kunye nemveliso yeasidi simahla kunye ammonia.I-Hydroxamic acids kunye nezinye i-asidi eziphilayo zisetyenziswa ngokubanzi njengeziyobisi kuba ziyinxalenye yezinto ezikhula, ii-antibiotics kunye ne-tumor inhibitors.I-amidases inokwahlulwa ibe yi-R kunye nohlobo lwe-acylases ngokwe-catalyst stereoelectivity.
Ukongeza kwi-catalyzing i-hydrolysis yee-amides, i-amidase inokuphinda ibangele ukusabela kokudluliselwa kwe-acyl kubukho be-co-substrates njenge-hydroxylamine.
I-Amidase enemithombo eyahlukeneyo ineenkalo ezahlukeneyo ze-substrate, ezinye zazo zinokwenza kuphela i-hydrolyze i-amides enuka kamnandi, ezinye zazo zinokwenza kuphela i-hydrolyze aliphatic amides, kunye ne-hydrolyze α-okanye ω-amino amides.Uninzi lwee-amines zinomsebenzi olungileyo we-catalytic kuphela kwi-acyclic okanye i-amides elula enuka kamnandi, kodwa kwi-aromatics entsonkothileyo, i-heterocyclic amides, ngakumbi i-amides ene-ortho substituents, idla ngokuba nomsebenzi ophantsi (zimbalwa kuphela ii-enzymes ezibonisa iziphumo ezingcono ze-catalytic).
Indlela yokwenza:
| Iienzymes | Ikhowudi yeMveliso | Ikhowudi yeMveliso |
| Umgubo we-Enzyme | ES-AMD-101~ ES-AMD-119 | iseti ye-19 amidases, 50 mg nganye 19 izinto * 50mg / into, okanye enye ubungakanani |
| Ikhithi yokuHlola (SynKit) | ES-AMD-1900 | iseti 19 amidases, 1 mg nganye 19 izinto * 1mg / into |
★ Ubucacileyo besubstrate ephezulu.
★ Ukukhetha okuqinileyo kwe-chiral.
★ uguqulo olusebenzayo.
★ Iimveliso ezingaphantsi kweemveliso.
★ Iimeko zokusabela ezithambileyo.
★ Ilungele indalo.
➢ Ukuhlolwa kwe-Enzyme kufuneka kuqhutywe kwi-substrates ezithile ngenxa ye-substrate ekhethekileyo, kwaye ufumane i-enzyme eyenza i-substrate ekujoliswe kuyo kunye neyona nto isebenzayo.
➢ Ungaze udibane neemeko ezigqithisileyo ezinje: ubushushu obuphezulu, i-pH ephezulu/ephantsi kunye ne-organic solvent ene-concentration ephezulu.
➢ Ngokuqhelekileyo, inkqubo yokusabela kufuneka ibandakanye i-substrate, isisombululo se-buffer (Eyona ndlela yokusabela i-pH ye-enzyme).Ii-co-substrates ezifana ne-hydroxylamine kufuneka zibekho kwi-acyl transfer reaction system.
➢ I-AMD kufuneka yongezwe okokugqibela kwisixokelelwano sokusabela nge-pH kunye nobushushu obuphezulu.
➢ Zonke iintlobo ze-AMD zineemeko ezahlukeneyo zokusabela, ngoko ke nganye kuzo kufuneka ifundwe ngakumbi ngayo.
Umzekelo 1(1):
Umsebenzi we-Hydrolysis kwii-Amide Substrates ezahlukeneyo
| Inxalenye engaphantsi | Umsebenzi othile μmols min-1mg-1 | Inxalenye engaphantsi | Umsebenzi othile μmols min-1mg-1 |
| IAcetamide | 3.8 | ο-OH benzamide | 1.4 |
| Ipropionamide | 3.9 | p-OH benzamide | 1.2 |
| ILactamide | 12.8 | ο-NH2benzamide | 1.0 |
| I-Butyramide | 11.9 | p-NH2benzamide | 0.8 |
| Isobutyramide | 26.2 | ο-Toluamide | 0.3 |
| I-Petanamide | 22.0 | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | INicotinamide | 1.7 |
| Cyclohexanamide | 19.5 | I-isonicotinamide | 1.8 |
| Acrylamide | 10.2 | I-Picolinamide | 2.1 |
| I-Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Iprolinamide | 3.4 | I-Indol-3-acetamide | 1.9 |
| I-Benzamide | 6.8 |
Ukusabela kwenziwa kwisisombululo se-50mM sesodium phosphate buffer, pH 7.5, ku-70 ℃.
| Amides | IHydroxylamine | IHydrazine |
| IAcetamide | 8.4 | 1.4 |
| Ipropionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| I-Benzamide | 9.2 | 6.1 |
Ukusabela kwenziwa kwisisombululo se-50mM sesodium phosphate buffer, pH 7.5, ku-70 ℃.
Ugxininiso lwe-reagent ehambelanayo: i-amides, i-100 mM (benzamide, i-10 mM);i-hydroxylamine kunye ne-hydrazine, i-400 mM;i-enzyme 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.Inkqubo ye-Biochemistry, i-2015, i-50 (8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al.I-Bioorganic Chemistry, ngo-2017, Ifumaneka kwi-intanethi 7.
