Amidase (AMD)
Enzima:O ni macromolecular biological catalysts, o le tele o enzymes o porotini
Amidase:Fa'agaoioi le hydrolysis o le tele o amides aliphatic ma aromatic endogenous ma fafo e ala i le fa'aliliuina o le acyl group i le vai fa'atasi ai ma le gaosiga o free acids ma le ammonia. O hydroxamic acids ma isi organic acids e fa'aaogaina lautele o ni vaila'au ona o ni vaega ia o mea e tutupu ai le tuputupu a'e, antibiotics ma tumor inhibitors. E mafai ona vaevaeina amidase i le ituaiga R ma le ituaiga S acylases e tusa ai ma le stereoselectivity o le catalyst.
I le fa'aopoopoga i le fa'agaoioiga o le hydrolysis o amides, e mafai fo'i e le amidase ona fa'agaoioiga ni tali o le acyl transfer i le i ai o co-substrates e pei o le hydroxylamine.
E eseese le fa'apitoa o substrate o le amidase ma punaoa eseese, o nisi o ia mea e na'o le hydrolyze o aromatic amides e mafai ona faia, o nisi e na'o le hydrolyze o aliphatic amides e mafai ona faia, ma o nisi e hydrolyze le α-po'o le ω-amino amides. O le tele o amines e lelei le catalytic activity mo na'o acyclic po'o simple aromatic amides, ae mo complex aromatics, o heterocyclic amides, aemaise lava amides ma ortho substituents, e masani ona maualalo le gaioiga (e na'o ni nai enzymes e sili atu ona lelei le catalytic activity).
Faiga fa'aolaola:
| Enzima | Fa'ailoga o le Oloa | Fa'ailoga o le Oloa |
| Pauta Enzyme | ES-AMD-101~ ES-AMD-119 | o se seti o amidase e 19, 50 mg i le seti 19 mea * 50mg / mea, po'o se isi lava aofa'i |
| Meafaigaluega mo le Su'esu'ega (SynKit) | ES-AMD-1900 | o se seti o amidase e 19, 1 mg i le seti 19 mea * 1mg / mea |
★ Maualuga le tulaga fa'apitoa o le substrate.
★ Filifiliga malosi o le chiral.
★ Lelei maualuga o le liua.
★ E itiiti ifo oloa e maua mai ai.
★ Tulaga o le tali atu e le ogaoga.
★ Fa'alelei i le siosiomaga.
➢ E tatau ona faia le su'esu'ega o le enzyme mo ni substrates faapitoa ona o le tulaga faapitoa o le substrate, ma maua ai se enzyme e fa'agaoioia le substrate fa'amoemoeina ma le aafiaga sili ona lelei.
➢ Aua lava ne'i pa'i atu i ni tulaga ogaoga e pei o le: vevela maualuga, pH maualuga/maualalo ma le organic solvent e maualuga le malosi.
➢ E masani lava, e tatau ona aofia i le faiga o le tali atu le substrate, le buffer solution (O le pH sili ona lelei o le tali atu o le enzyme). E tatau ona iai ni co-substrates e pei o le hydroxylamine i le faiga o le tali atu o le acyl transfer.
➢ E tatau ona fa'aopoopo mulimuli le AMD i totonu o le faiga o tali fa'atasi ai ma le pH ma le vevela sili ona lelei o le tali.
➢ O ituaiga uma o AMD e iai tulaga eseese o le tali atu sili ona lelei, o lea e tatau ai ona suʻesuʻeina atili tulaga taʻitasi.
Faʻataʻitaʻiga 1(1):
Gaoioiga o le hydrolysis i Amide Substrates eseese
| Substrate | Gaoioiga faapitoa μmols min-1mg-1 | Substrate | Gaoioiga faapitoa μmols min-1mg-1 |
| Asetamida | 3.8 | ο-OH benzamide | 1.4 |
| Propionamide | 3.9 | p-OH benzamide | 1.2 |
| Lactamide | 12.8 | ο-NH2benzamida | 1.0 |
| Butyramide | 11.9 | p-NH2benzamida | 0.8 |
| Isobutyramide | 26.2 | ο-Toluamide | 0.3 |
| Pentanamide | 22.0 | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | Nikotinamida | 1.7 |
| Sikloheksanamida | 19.5 | Isonicotinamide | 1.8 |
| Akrilamida | 10.2 | Pikolinamide | 2.1 |
| Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Prolinamide | 3.4 | Indol-3-acetamide | 1.9 |
| Benzamide | 6.8 |
Sa faia le tali atu i le 50mM sodium phosphate buffer solution, pH 7.5, i le 70 ℃.
| Amides | Hydroxylamine | Hydrazine |
| Asetamida | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
Sa faia le tali atu i le 50mM sodium phosphate buffer solution, pH 7.5, i le 70 ℃.
O le malosi o le reagent e feso'ota'i i ai: amides, 100 mM (benzamide, 10 mM); hydroxylamine ma le hydrazine, 400 mM; enzyme 0.9 μM.
Fa'ata'ita'iga 2(2):
Fa'ata'ita'iga 3(3):
1. D'Abusco AS, Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, ma isi. Process Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, ma isi. Kemisi Fa'aolaola, 2017, Maua i luga ole initaneti 7.
