Amidase (AMD)
Ngā whākōkī:He whakakorikori koiora macromolecular, ko te nuinga o ngā whākōkī he pūmua
Āmidase:Whakakorikori i te waikura o ngā momo amide aliphatic me te kakara taketake me ngā amide tauhou mā te whakawhiti i tētahi rōpū acyl ki te wai me te whakaputanga o ngā waikawa kore utu me te haukini. He whānuitia te whakamahinga o ngā waikawa hydroxamic me ētahi atu waikawa rauropi hei rongoā nā te mea he wāhanga rātou o ngā mea whakatipu, ngā paturopi me ngā aukati pukupuku. Ka taea te wehewehe i ngā amidase ki ngā acylases momo R me te momo S e ai ki te stereoselectivity o te whakakorikori.
Haunga te whakateretere i te waikura o ngā amides, ka taea hoki e te amidase te whakateretere i ngā tauhohenga whakawhiti acyl i te aroaro o ngā matū-tahi pērā i te hydroxylamine.
He rerekē te motuhaketanga o ngā matūriki o ngā amidase me ngā pūtake rerekē, ko ētahi o rātou ka taea anake te whakawhaiti i ngā amide kakara, ko ētahi ka taea anake te whakawhaiti i ngā amide aliphatic, ā, ko ētahi ka whakawhaiti i ngā amide α-, ω-amino rānei. He pai te mahi whakakorikori a te nuinga o ngā amine mō ngā amide kakara acyclic, ngā amide kakara māmā rānei, engari mō ngā kakara matatini, he iti te mahi a ngā amide heterocyclic, inā koa ngā amide me ngā whakakapinga ortho (he torutoru noa iho ngā whākōkī e whakaatu ana i ngā pānga whakakorikori pai ake).
Tikanga whakaterenga:
| Ngā whākōkī | Waehere Hua | Waehere Hua |
| Paura whākōkī | ES-AMD-101~ ES-AMD-119 | he huinga o ngā amidase 19, 50 mg ia 19 ngā tūemi * 50mg/ tūemi, tētahi atu rahinga rānei |
| Kete Tirotiro (SynKit) | ES-AMD-1900 | he huinga o ngā amidase 19, 1 mg ia 19 ngā tūemi * 1mg / tūemi |
★ Teitei o te motuhaketanga o te papa.
★ He kaha te whiriwhiringa chiral.
★ Te tino whai huatanga o te tahuritanga.
★ He iti ake ngā hua taha.
★ Ngā āhuatanga tauhohenga ngawari.
★ He pai mō te taiao.
➢ Me whakahaere te tātari whākōkī mō ngā matū motuhake nā te mea he motuhake te matū, ā, me tiki he whākōkī e whakaoho ana i te matū ūnga me te pānga whakaoho pai rawa atu.
➢ Kaua rawa e pā ki ngā āhuatanga tino kino pēnei i te: pāmahana teitei, pH teitei/iti me te whakarewa waro me te kukū teitei.
➢ I te nuinga o te wā, me whakauru te pūnaha tauhohenga i te matūriki, te otinga ārai (te pH tauhohenga tino pai o te whākōkī). Me noho ngā matūriki-tahi pēnei i te hydroxylamine ki roto i te pūnaha tauhohenga whakawhiti acyl.
➢ Me tāpiri whakamutunga te AMD ki te pūnaha tauhohenga me te pH me te pāmahana tauhohenga tino pai.
➢ He rerekē ngā āhuatanga tauhohenga tino pai o ngā momo AMD katoa, nō reira me ako anō ia āhuatanga takitahi.
Tauira 1(1):
Te mahi waikura ki ngā papa amide rerekē
| Papa raro | Mahi motuhake μmols min-1mg-1 | Papa raro | Mahi motuhake 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 |
| Pūtirimiti | 11.9 | p-NH2benzamida | 0.8 |
| Isobutyramide | 26.2 | ο-Toluamide | 0.3 |
| Pentanamide | 22.0 | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | Nikotīmida | 1.7 |
| Sikorohekanamida | 19.5 | Isonicotinamide | 1.8 |
| Akiriramiti | 10.2 | Pikolinamida | 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 buffer, pH 7.5, i te 70 ℃.
| Ngā Amides | Hydroxylamine | Hydrazine |
| Asetamida | 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 buffer, pH 7.5, i te 70 ℃.
Te kukū o te matū matū e pā ana: ngā amide, 100 mM (benzamide, 10 mM); hydroxylamine me te hydrazine, 400 mM; te 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. Te Matūkai Tukanga, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al. Matū Pūtaiao Koiora, 2017, Kei te wātea i te ipurangi 7.
