Amidase (AMD)
Enzymes:Yog macromolecular lom catalysts, feem ntau enzymes yog cov proteins
Amidase:Catalyze lub hydrolysis ntawm ntau yam endogenous thiab txawv teb chaws aliphatic thiab aromatic amides los ntawm kev hloov ib pawg acyl rau dej nrog zus tau tej cov acids dawb thiab ammonia.Hydroxamic acids thiab lwm cov organic acids tau dav siv los ua tshuaj vim tias lawv yog cov khoom siv ntawm kev loj hlob, tshuaj tua kab mob thiab qog nqaij hlav.Cov amidases tuaj yeem muab faib ua R hom thiab S hom acylases raws li cov catalyst stereoselectivity.
Ntxiv nrog rau catalyzing hydrolysis ntawm amides, amidase kuj tseem tuaj yeem ua rau cov kev hloov pauv acyl nyob rau hauv muaj cov co-substrates xws li hydroxylamine.
Amidase nrog ntau qhov chaw muaj qhov sib txawv ntawm cov substrate, qee qhov ntawm lawv tuaj yeem tsuas yog hydrolyze aromatic amides, qee tus tuaj yeem tsuas yog hydrolyze aliphatic amides, thiab qee qhov hydrolyze α-lossis ω-amino amides.Feem ntau ntawm cov amines muaj kev ua haujlwm zoo catalytic nkaus xwb rau acyclic lossis yooj yim aromatic amides, tab sis rau complex aromatics, heterocyclic amides, tshwj xeeb tshaj yog amides nrog ortho hloov, feem ntau tsis tshua muaj kev ua si (tsuas yog ob peb enzymes pom zoo catalytic nyhuv).
Catalytic mechanism:
| Enzymes | Khoom Code | Khoom Code |
| Enzyme hmoov | ES-AMD-101 ~ ES-AMD-119 | ib txheej ntawm 19 amidases, 50 mg txhua 19 yam * 50mg / yam khoom, lossis lwm yam |
| Cov khoom siv tshuaj ntsuam xyuas (SynKit) | ES-AMD-1900 | ib txheej ntawm 19 amidases, 1 mg txhua 19 yam khoom * 1mg / khoom |
★ High substrate specification.
★ Muaj zog chiral selectivity.
★ High conversion efficiency.
★ Tsawg los ntawm cov khoom lag luam.
★ Cov tshuaj tiv thaiv me me.
★ Ib puag ncig tus phooj ywg.
➢ Kev tshuaj xyuas Enzyme yuav tsum tau ua rau cov substrates tshwj xeeb vim yog cov substrate tshwj xeeb, thiab tau txais ib qho enzyme uas catalyzes lub hom phiaj substrate nrog cov nyhuv catalytic zoo tshaj plaws.
➢ Tsis txhob tiv tauj nrog cov xwm txheej hnyav xws li: qhov kub thiab txias, pH siab / qis thiab cov kuab tshuaj organic nrog siab concentration.
➢ Nquag, cov txheej txheem tshuaj tiv thaiv yuav tsum suav nrog substrate, tsis muaj tshuaj (Cov tshuaj tiv thaiv zoo pH ntawm enzyme).Co-substrates xws li hydroxylamine yuav tsum muaj nyob rau hauv acyl hloov cov tshuaj tiv thaiv kab mob.
➢ AMD yuav tsum tau ntxiv qhov kawg rau hauv cov tshuaj tiv thaiv nrog cov tshuaj tiv thaiv zoo pH thiab kub.
➢ Txhua yam ntawm AMD muaj ntau yam kev pom zoo, yog li txhua tus ntawm lawv yuav tsum tau kawm ntxiv ib tus zuj zus.
Piv txwv 1(1):
Hydrolysis kev ua si rau txawv Amide Substrates
| Substrate | Cov haujlwm tshwj xeeb hli min-1mg-1 | Substrate | Cov haujlwm tshwj xeeb hli min-1mg-1 |
| Acetamide | 3.8 | ο- OH benzamide yog | 1.4 |
| Propionamide | 3.9 | p- OH benzamide yog | 1.2 |
| Lactamide | 12.8 ib | ο-NH2tshuaj benzamide | 1.0 |
| Butyramide | 11.9 ib | p-NH2tshuaj benzamide | 0.8 ua |
| Isobutyramide | 26.2 ib | ο-Toluamide | 0.3 |
| Pentanamide | 22.0 Nws | p-Toluamide | 8.1 |
| Hexanamide | 6.4 | Nicotinamide | 1.7 |
| Cyclohexanamide | 19.5 Nws | 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 nyob rau hauv 50mM sodium phosphate buffer tov, pH 7.5, ntawm 70 ℃.
| Amides | Hydroxylamine | Hydrazine |
| Acetamide | 8.4 | 1.4 |
| Propionamide | 18.4 ib | 3.0 |
| Isobutyramide | 25.0 Nws | 22.7 ib |
| Benzamide | 9.2 ib | 6.1 |
Cov tshuaj tiv thaiv tau ua nyob rau hauv 50mM sodium phosphate buffer tov, pH 7.5, ntawm 70 ℃.
Muaj feem xyuam reagent concentration: 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, et al.Txheej Txheem Biochemistry, 2015, 50(8): 1400-1404.
3. Zheng RC, Jin JQ, Wu ZM, et al.Bioorganic Chemistry, 2017, Muaj nyob online 7.
