Amidase (AMD)
Enzymes:He whakakōkī koiora macromolecular, he pūmua te nuinga o ngā whākōkī
Amidase:Whakakotahitia te hauwai o nga momo amides aliphatic me nga amide kakara mai kee ma te whakawhiti i tetahi roopu acyl ki te wai me te hanga waikawa kore utu me te haukini.Ko nga waikawa Hydroxamic me etahi atu waikawa waro e whakamahia nuitia ana hei raau taero na te mea ko enei nga waahanga o te tipu, nga paturopi me te aukati puku.Ka taea te wehea nga amidases ki te momo R me te momo S acylases e ai ki te stereoselectivity catalyst.
I tua atu i te whakakorikori i te hydrolysis o nga amides, ka taea ano e te amidase te whakakorikori i nga tauhohenga whakawhiti acyl i te aroaro o nga taapiri penei i te hydroxylamine.
Ko te Amidase me nga punaa rereke he rereke te motuhake o te tïpako, ka taea e etahi o ratou te whakamaarama noa i nga amida kakara, ko etahi ka taea anake te whakamaarama i nga amida aliphatic, ko etahi o nga amida α-, ω-amino ranei.Ko te nuinga o nga amine he pai te mahi catalytic mo nga amida kakara acyclic, ngawari noa iho ranei, engari mo nga mea kakara uaua, ko nga amida heterocyclic, ina koa ko nga amides whai ortho substituent, he iti noa te mahi (he iti noa nga enzyme e whakaatu ana i nga paanga catalytic pai ake).
Hangahanga catalytic:
| Nga Enzymes | Waehere Hua | Waehere Hua |
| Te paura Enzyme | ES-AMD-101~ ES-AMD-119 | he huinga o 19 amidases, 50 mg ia 19 tuemi * 50mg / taonga, etahi atu rahinga ranei |
| Kete Tirohanga (SynKit) | ES-AMD-1900 | he huinga o 19 amidases, 1 mg ia 19 tūemi * 1mg / tūemi |
★ motuhake tïpako High.
★ He kaha te whiriwhiringa o te chiral.
★ te kaha o te hurihanga teitei.
★ He iti ake nga hua.
★ Nga tikanga tauhohenga ngawari.
★ He pai te taiao.
➢ Me mahi te tirotiro i te haukini mo nga taputapu motuhake na te mea he motuhake te taputapu, me te tiki i tetahi whaariki e whakakorikori ana i te tïpako i whäia me te whai hua whakatiki pai rawa atu.
➢ Kaua rawa e pa atu ki nga ahuatanga kino penei: te pāmahana teitei, te pH teitei/iti me te whakarewa pararopi me te kukū teitei.
➢ Ko te tikanga, me whakauru te punaha tauhohenga he tïpako, he wairewa buffer (Ko te pH tauhohenga tino pai o te enzyme).Me noho tahi nga taarua penei i te hydroxylamine ki te punaha tauhohenga whakawhiti acyl.
➢ Me whakauru te AMD ki te punaha tauhohenga me te pH tauhohenga tino pai me te pāmahana.
➢ Ko nga momo AMD katoa he rerekee nga tikanga tauhohenga tino pai, no reira me ako takitahi ia o aua momo.
Tauira 1(1):
Te mahi waiwai ki nga Raupapa Amide rereke
| Te papanga | Mahi motuhake μmols min-1mg-1 | Te papanga | Mahi motuhake μmols min-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 | Pikolinamide | 2.1 |
| Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Prolinamide | 3.4 | Indol-3-acetamide | 1.9 |
| Benzamide | 6.8 |
I mahia te tauhohenga i roto i te 50mM konutai phosphate wairewa buffer, pH 7.5, i te 70 ℃.
| Amides | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
I mahia te tauhohenga i roto i te 50mM konutai phosphate wairewa buffer, pH 7.5, i te 70 ℃.
Te kukū tauhohenga e pa ana: amides, 100 mM(benzamide, 10 mM);hydroxylamine me te hydrazine, 400 mM;whākōkī 0.9 μM.
Tauira 2(2):
Tauira 3(3):
1. D'Abusco AS, Ammendola S., et al.Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, et al.Tukatuka Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al.Bioorganic Chemistry, 2017, Kei runga ipurangi 7.
