Amidase (AMD)
Cov Enzymes:Yog cov catalysts macromolecular biological, feem ntau cov enzymes yog cov proteins
Amidase:Ua kom cov hydrolysis ntawm ntau yam endogenous thiab txawv teb chaws aliphatic thiab aromatic amides los ntawm kev hloov ib pawg acyl mus rau hauv dej nrog rau kev tsim cov kua qaub dawb thiab ammonia. Hydroxamic acids thiab lwm yam organic acids yog siv dav ua tshuaj vim tias lawv yog cov khoom xyaw ntawm cov yam tseem ceeb loj hlob, tshuaj tua kab mob thiab cov tshuaj tiv thaiv qog. Cov amidases tuaj yeem faib ua R hom thiab S hom acylases raws li catalyst stereoselectivity.
Ntxiv rau qhov ua rau lub hydrolysis ntawm amides, amidase kuj tseem tuaj yeem ua rau cov tshuaj hloov pauv acyl hloov pauv nyob rau hauv qhov muaj cov co-substrates xws li hydroxylamine.
Amidase nrog cov peev txheej sib txawv muaj qhov tshwj xeeb ntawm cov substrate sib txawv, qee qhov ntawm lawv tsuas yog hydrolyze aromatic amides, qee qhov ntawm lawv tsuas yog hydrolyze aliphatic amides, thiab qee qhov hydrolyze α-lossis ω-amino amides. Feem ntau ntawm cov amines muaj cov haujlwm catalytic zoo tsuas yog rau acyclic lossis yooj yim aromatic amides, tab sis rau cov aromatics nyuaj, heterocyclic amides, tshwj xeeb tshaj yog amides nrog ortho substituents, feem ntau yog qis hauv kev ua haujlwm (tsuas yog ob peb lub enzymes qhia txog cov teebmeem catalytic zoo dua).
Cov txheej txheem catalytic:
| Cov Enzymes | Cov Cai Khoom | Cov Cai Khoom |
| Hmoov Enzyme | ES-AMD-101~ ES-AMD-119 | ib pawg ntawm 19 amidases, 50 mg txhua 19 yam khoom * 50mg / yam khoom, lossis lwm yam ntau npaum li cas |
| Cov Khoom Siv Tshuaj Ntsuam Xyuas (SynKit) | ES-AMD-1900 | ib pawg ntawm 19 amidases, 1 mg txhua yam 19 yam khoom * 1mg / yam khoom |
★ Kev tshwj xeeb ntawm cov substrate siab.
★ Muaj zog xaiv chiral.
★ Kev hloov pauv siab heev.
★ Cov khoom tsis muaj txiaj ntsig tsawg dua.
★ Cov mob ua xua me me.
★ Ib puag ncig zoo.
➢ Yuav tsum tau kuaj xyuas enzyme rau cov substrates tshwj xeeb vim yog qhov tshwj xeeb ntawm cov substrate, thiab tau txais cov enzyme uas ua rau lub substrate catalytic zoo tshaj plaws.
➢ Tsis txhob kov cov xwm txheej hnyav xws li: kub heev, pH siab/qis thiab cov kuab tshuaj organic uas muaj concentration siab.
➢ Feem ntau, lub kaw lus tshuaj tiv thaiv yuav tsum muaj cov substrate, buffer solution (Qhov pH zoo tshaj plaws ntawm cov enzyme). Cov co-substrates xws li hydroxylamine yuav tsum muaj nyob rau hauv lub kaw lus tshuaj tiv thaiv acyl.
➢ Yuav tsum ntxiv AMD rau hauv qhov system tshuaj tiv thaiv uas muaj pH thiab kub zoo tshaj plaws.
➢ Txhua hom AMD muaj ntau yam kev mob tshwm sim zoo tshaj plaws, yog li txhua yam ntawm lawv yuav tsum tau kawm ntxiv.
Piv txwv 1(1):
Kev ua haujlwm hydrolysis rau ntau yam Amide Substrates
| Cov khoom siv hauv qab | Kev ua ub no tshwj xeeb μmols min-1mg-1 | Cov khoom siv hauv qab | Kev ua ub no tshwj xeeb 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 | Picolinamide | 2.1 |
| Metacrylamide | 3.5 | 3-Phenylpropionamide | 7.6 |
| Prolinamide | 3.4 | Indol-3-acetamide | 1.9 |
| Benzamide | 6.8 |
Cov tshuaj tiv thaiv tau ua tiav hauv 50mM sodium phosphate buffer solution, pH 7.5, ntawm 70 ℃.
| Cov tshuaj amide | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| Propionamide | 18.4 | 3.0 |
| Isobutyramide | 25.0 | 22.7 |
| Benzamide | 9.2 | 6.1 |
Cov tshuaj tiv thaiv tau ua tiav hauv 50mM sodium phosphate buffer solution, pH 7.5, ntawm 70 ℃.
Qhov concentration ntawm cov reagent: amides, 100 mM (benzamide, 10 mM); hydroxylamine thiab hydrazine, 400 mM; enzyme 0.9 μM.
Piv txwv 2(2):
Piv txwv 3(3):
1. D'Abusco AS, Ammendola S., et al. Extremophiles, 2001, 5:183-192.
2. Guo FM, Wu JP, Yang LR, thiab lwm tus. Txheej Txheem Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, thiab lwm tus. Bioorganic Chemistry, 2017, Muaj nyob hauv online 7.
