Amidase (AMD)
Nā Enzymes:ʻO ka macromolecular biological catalysts, ʻo ka hapa nui o nā enzyme he protein
Amidase:Hoʻopili i ka hydrolysis o nā endogenous a me nā haole aliphatic a me nā amides aromatic ma o ka hoʻololi ʻana i kahi hui acyl i ka wai me ka hana ʻana o nā ʻakika manuahi a me ka amonia.Hoʻohana nui ʻia nā ʻakika Hydroxamic a me nā mea ʻokoʻa ʻē aʻe e like me nā lāʻau lapaʻau no ka mea he mau mea ia o nā kumu ulu, nā lāʻau antibiotic a me nā mea hoʻopaneʻe tumor.Hiki ke hoʻokaʻawale ʻia nā amidases i R type a me S type acylases e like me ka stereoselectivity catalyst.
Ma waho aʻe o ka hoʻopili ʻana i ka hydrolysis o nā amides, hiki nō i ka amidase ke hoʻololi i nā hopena acyl transfer i mua o nā co-substrates e like me hydroxylamine.
ʻO ka Amidase me nā kumu like ʻole he substrate specificity, hiki i kekahi o lākou ke hydrolyze aromatic amides wale nō, hiki i kekahi o lākou ke hydrolyze aliphatic amides wale nō, a me kekahi hydrolyze α-a i ʻole ω-amino amides.ʻO ka hapa nui o nā amine he hana catalytic maikaʻi wale nō no nā amides acyclic a maʻalahi paha, akā no nā mea ʻala paʻakikī, heterocyclic amides, ʻoi aku ka nui o nā amides me nā ortho substituents, he haʻahaʻa i ka hana (he mau enzymes wale nō e hōʻike ana i nā hopena catalytic maikaʻi aʻe).
ʻO ka mīkini hoʻoheheʻe:
| Nā Enzymes | Code Huahana | Code Huahana |
| ʻO ka pauka Enzyme | ES-AMD-101~ ES-AMD-119 | he set o 19 amidases, 50 mg i kēlā me kēia 19 mau mea * 50mg / mea, a i ʻole ka nui ʻē aʻe |
| Mea Hoʻomaʻamaʻa (SynKit) | ES-AMD-1900 | he set o 19 amidases, 1 mg kēlā me kēia 19 mau mea * 1mg / mea |
★ High substrate specificity.
★ Ke koho kiral ikaika.
★ kiʻekiʻe hoʻololi kūpono.
★ Liʻiliʻi nā huahana.
★ Nā kūlana hoʻonā haʻahaʻa.
★ Pili kaiapuni.
➢ Pono e hoʻokō ʻia ka nānā ʻana i ka enzyme no nā substrate kikoʻī ma muli o ke ʻano kikoʻī o ka substrate, a e kiʻi i kahi enzyme e hoʻolalelale i ka substrate i manaʻo ʻia me ka hopena catalytic maikaʻi loa.
➢ Mai hoʻopili aku me nā kūlana koʻikoʻi e like me: ka wela kiʻekiʻe, ka pH kiʻekiʻe/haʻahaʻa a me ka mea hoʻoheheʻe organik me ka ʻike kiʻekiʻe.
➢ ʻO ka mea maʻamau, pono e hoʻokomo i ka ʻōnaehana hoʻololi i ka substrate, ka solution buffer (ʻO ka pH hopena maikaʻi loa o ka enzyme).Pono nā mea hoʻohui like e like me ka hydroxylamine i loko o ka ʻōnaehana hoʻololi acyl.
➢ Pono e hoʻohui hope ʻia ka AMD i loko o ka ʻōnaehana pane me ka pH hopena maikaʻi loa a me ka mahana.
➢ Loaʻa i nā ʻano AMD a pau ke ʻano o ka hopena maikaʻi loa, no laila pono e aʻo hou ʻia kēlā me kēia o lākou.
Laʻana 1(1):
ʻO ka hana hydrolysis i nā substrate Amide like ʻole
| Pāpaʻa | Hana kūikawā μmols min-1mg-1 | Pāpaʻa | Hana kūikawā μmols min-1mg-1 |
| Acetamide | 3.8 | ο-OH benzamide | 1.4 |
| ʻO 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 | ʻO Isonicotinamide | 1.8 |
| Acrylamide | 10.2 | Picolinamide | 2.1 |
| Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Prolinamide | 3.4 | Indol-3-acetamide | 1.9 |
| Benzamide | 6.8 |
Ua hoʻokō ʻia ka hopena ma 50mM sodium phosphate buffer solution, pH 7.5, ma 70 ℃.
| Amides | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| ʻO propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
Ua hoʻokō ʻia ka hopena ma 50mM sodium phosphate buffer solution, pH 7.5, ma 70 ℃.
ʻO ka hoʻopaʻa ʻana o ka reagent pili: amides, 100 mM(benzamide, 10 mM);hydroxylamine a me ka 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, a me al.Kaʻina Biochemistry, 2015, 50 (8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al.Bioorganic Chemistry, 2017, Loaʻa ma ka pūnaewele 7.
