Amidase (AMD)
Ma enzyme:Ndi macromolecular biological catalysts, ma enzyme ambiri ndi mapuloteni
Pakati pa:Konzani hydrolysis ya ma aliphatic ndi aromatic amides osiyanasiyana amkati ndi akunja mwa kusamutsa gulu la acyl kupita kumadzi ndikupanga ma free acid ndi ammonia. Ma hydroxamic acid ndi ma organic acid ena amagwiritsidwa ntchito kwambiri ngati mankhwala chifukwa ndi omwe amapanga zinthu zokulitsa, maantibayotiki ndi zoletsa zotupa. Ma amidases amatha kugawidwa m'magulu a R type ndi S type acylases malinga ndi catalyst stereoselectivity.
Kuwonjezera pa kuyambitsa hydrolysis ya amides, amidase imathanso kuyambitsa acyl transfer reactions pamaso pa ma co-substrates monga hydroxylamine.
Ma amidase okhala ndi magwero osiyanasiyana ali ndi mawonekedwe osiyana a substrate, ena mwa iwo amatha kupopera amide a aromatic, ena amatha kupopera aliphatic amide, ndipo ena amatha kupopera α-kapena ω-amino amide. Ambiri mwa ma amines ali ndi ntchito yabwino yokonza ma catalytic kokha kwa acyclic kapena simple aromatic amides, koma kwa ma aromatic ovuta, ma heterocyclic amides, makamaka ma amide okhala ndi ortho substituents, nthawi zambiri amakhala ndi ntchito yochepa (ma enzyme ochepa okha ndi omwe amasonyeza zotsatira zabwino zokonza ma catalytic).
Njira yothandizira:
| Ma enzyme | Khodi ya Zamalonda | Khodi ya Zamalonda |
| Ufa wa Enzyme | ES-AMD-101~ ES-AMD-119 | seti ya 19 amidases, 50 mg iliyonse 19 zinthu * 50mg / chinthu, kapena kuchuluka kwina |
| Zida Zowunikira (SynKit) | ES-AMD-1900 | seti ya 19 amidases, 1 mg iliyonse Zinthu 19 * 1mg / chinthu |
★ Kuchuluka kwa substrate.
★ Kusankha bwino kwa chiral.
★ Kutembenuza bwino kwambiri.
★ Zogulitsa zochepa.
★ Zinthu zofatsa zomwe zimachitika chifukwa cha kukwiya.
★ Yosamalira chilengedwe.
➢ Kuwunika ma enzyme kuyenera kuchitika pa ma substrate enaake chifukwa cha kudziwika kwa substrate, ndikupeza enzyme yomwe imayambitsa substrate yomwe ikufunidwa ndi zotsatira zabwino kwambiri zoyambitsa.
➢ Musagwirizane ndi zinthu zoopsa monga: kutentha kwambiri, pH yapamwamba/yotsika komanso organic solvent yokhala ndi kuchuluka kwakukulu.
➢ Kawirikawiri, dongosolo la reaction liyenera kukhala ndi substrate, buffer solution (PH yabwino kwambiri ya reaction ya enzyme). Ma co-substrates monga hydroxylamine ayenera kukhalapo mu dongosolo la acyl transfer reaction.
➢ AMD iyenera kuwonjezeredwa komaliza mu dongosolo la reaction lomwe lili ndi pH yabwino kwambiri ya reaction ndi kutentha.
Mitundu yonse ya AMD ili ndi mikhalidwe yosiyanasiyana yabwino yochitira zinthu, kotero iliyonse iyenera kuphunziridwa payekhapayekha.
Chitsanzo 1(1):
Ntchito ya hydrolysis ku ma substrates osiyanasiyana a Amide
| Pansi pa nthaka | Zochita zapadera μmols mphindi-1mg-1 | Pansi pa nthaka | Zochita zapadera μmols mphindi-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 | Nikotinamide | 1.7 |
| Cyclohexanamide | 19.5 | 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 |
Yankho linachitika mu yankho la sodium phosphate buffer la 50mM, pH 7.5, pa 70 ℃.
| Amide | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
Yankho linachitika mu yankho la sodium phosphate buffer la 50mM, pH 7.5, pa 70 ℃.
Kuchuluka kwa reagent komwe kumagwirizana: amides, 100 mM(benzamide, 10 mM); hydroxylamine ndi hydrazine, 400 mM; enzyme 0.9 μM.
Chitsanzo 2(2):
Chitsanzo 3(3):
1. D'Abusco AS, Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, ndi ena. Process Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, ndi ena. Bioorganic Chemistry, 2017, Ikupezeka pa intaneti 7.
