ʻAmidase (AMD)
Nā ʻenzyme:He mau mea hoʻouluulu olaola macromolecular, ʻo ka hapa nui o nā enzymes he mau protein
ʻAmidase:Hoʻoulu i ka hydrolysis o nā ʻano endogenous a me nā ʻano aliphatic a me nā ʻala ʻē aʻe ma ka hoʻoili ʻana i kahi hui acyl i ka wai me ka hana ʻana o nā waikawa manuahi a me ka ammonia. Hoʻohana nui ʻia nā waikawa Hydroxamic a me nā waikawa organik ʻē aʻe ma ke ʻano he lāʻau lapaʻau no ka mea he mau ʻāpana lākou o nā mea ulu, nā antibiotics a me nā mea hoʻopaʻa tumor. Hiki ke hoʻokaʻawale ʻia nā amidase i nā ʻano R a me nā ʻano S acylases e like me ka stereoselectivity catalyst.
Ma waho aʻe o ka hoʻoulu ʻana i ka hydrolysis o nā amides, hiki i ka amidase ke hoʻoulu i nā hopena hoʻoili acyl i ke alo o nā co-substrates e like me hydroxylamine.
He ʻokoʻa ke ʻano o ka substrate o nā amidase me nā kumu like ʻole, hiki i kekahi o lākou ke hydrolyze i nā amides aromatic wale nō, hiki i kekahi o lākou ke hydrolyze i nā amides aliphatic wale nō, a ʻo kekahi e hydrolyze i nā amides α-a i ʻole ω-amino. Loaʻa i ka hapa nui o nā amines ka hana catalytic maikaʻi no nā amides aromatic acyclic a maʻalahi paha, akā no nā aromatics paʻakikī, ʻo nā amides heterocyclic, ʻoi aku ka amides me nā ortho substituents, he haʻahaʻa ka hana (he kakaikahi wale nō nā enzymes e hōʻike ana i nā hopena catalytic maikaʻi aʻe).
ʻAno hana hoʻouluulu:
| Nā ʻenzyme | Kōmike Huahana | Kōmike Huahana |
| Pauka ʻenzyme | ES-AMD-101~ ES-AMD-119 | he hoʻonohonoho o 19 amidase, 50 mg kēlā me kēia 19 mau mea * 50mg / mea, a i ʻole ka nui ʻē aʻe |
| Pahu Nānā (SynKit) | ES-AMD-1900 | he hoʻonohonoho o 19 amidase, 1 mg kēlā me kēia 19 mau mea * 1mg / mea |
★ Kiʻekiʻe ke ʻano kikoʻī o ka substrate.
★ Koho chiral ikaika.
★ Ka pono hoʻololi kiʻekiʻe.
★ Emi nā huahana ʻaoʻao.
★ Nā kūlana pane iki.
★ Mālama i ke kaiapuni.
➢ Pono e hoʻokō ʻia ka nānā ʻana o nā enzyme no nā substrates kikoʻī ma muli o ke ʻano kikoʻī o ka substrate, a loaʻa i kahi enzyme e hoʻoulu ai i ka substrate i manaʻo ʻia me ka hopena catalytic maikaʻi loa.
➢ Mai hoʻopā aku me nā kūlana koʻikoʻi e like me: ke kiʻekiʻe o ka mahana, ka pH kiʻekiʻe/haʻahaʻa a me ka mea hoʻoheheʻe organik me ka nui kiʻekiʻe.
➢ ʻO ka maʻamau, pono i ka ʻōnaehana hana ke komo pū me ka substrate, ka hopena buffer (ʻO ka pH hopena kūpono o ka enzyme). Pono nā co-substrates e like me ka hydroxylamine e kū i loko o ka ʻōnaehana hana hoʻoili acyl.
➢ Pono e hoʻohui hope ʻia ka AMD i loko o ka ʻōnaehana hana me ka pH a me ka mahana hana kūpono loa.
➢ Loaʻa i nā ʻano AMD āpau nā kūlana hopena kūpono like ʻole, no laila pono e aʻo hou ʻia kēlā me kēia.
Laʻana 1(1):
Hana hydrolysis i nā ʻano Amide Substrates like ʻole
| Pākuʻi | Hana kikoʻī μmols min-1mg-1 | Pākuʻi | Hana kikoʻī μmols min-1mg-1 |
| ʻAsetamida | 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 |
| ʻIsobutyramide | 26.2 | ο-Toluamide | 0.3 |
| Pentanamide | 22.0 | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | ʻO ka nikotinamida | 1.7 |
| Sikloheksanamida | 19.5 | ʻIsonicotinamide | 1.8 |
| ʻAkrilamida | 10.2 | Pikolinamide | 2.1 |
| ʻO ka Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| ʻO Prolinamide | 3.4 | ʻO Indol-3-acetamide | 1.9 |
| ʻO Benzamide | 6.8 |
Ua hoʻokō ʻia ka hopena ma ka 50mM sodium phosphate buffer solution, pH 7.5, ma 70 ℃.
| ʻAmides | ʻO ka Hydroxylamine | ʻO ka Hydrazine |
| ʻAsetamida | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| ʻIsobutyramide | 25.0 | 22.7 |
| ʻO Benzamide | 9.2 | 6.1 |
Ua hoʻokō ʻia ka hopena ma ka 50mM sodium phosphate buffer solution, pH 7.5, ma 70 ℃.
Ka nui o ka reagent e pili ana: amides, 100 mM (benzamide, 10 mM); hydroxylamine a me hydrazine, 400 mM; enzyme 0.9 μM.
Laʻana 2(2):
Laʻana 3(3):
1. D'Abusco AS, Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, et al. Kaʻina Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al. Kemika Bioorganic, 2017, Loaʻa ma ka pūnaewele 7.
