I-4-Nitrophenyl beta-D-galactopyranoside CAS:200422-18-0

Isiphumo: I-ONPG yi-substrate esetyenziselwa ngokukodwa ukubona ubukho kunye nomsebenzi we-enzyme i-β-galactosidase. Xa i-enzyme ye-β-galactosidase ikhona kwaye isebenza, iqhekeza i-ONPG ibe ziimveliso ezimbini: i-o-nitrophenol kunye ne-galactose derivative. Ukukhululwa kwe-o-nitrophenol kubangela utshintsho lombala otyheli, olunokulinganiswa kusetyenziswa i-spectrophotometer.

Isicelo: I-ONPG inezicelo ezininzi kuphando lwebhayoloji yemolekyuli kunye ne-biochemistry:

Ukuchonga umsebenzi we-β-galactosidase: I-ONPG isetyenziswa rhoqo ukulinganisa nokulinganisa umsebenzi we-enzyme ye-β-galactosidase. Izinga lokwakheka kwe-o-nitrophenol, elihambelana ngqo nomsebenzi we-enzyme, linokulinganiswa nge-spectrophotometric.

Ukubonakaliswa kwezakhi zofuzo kunye nolawulo: I-ONPG idla ngokusetyenziswa kwiimvavanyo ezinxulumene nokubonakaliswa kwezakhi zofuzo kunye nezifundo zolawulo. Isetyenziswa rhoqo kwiimvavanyo zeprotheyini yokudibanisa, njengenkqubo yokuhlanganiswa kwe-lacZ esetyenziswa rhoqo, ukufunda ukubonakaliswa kwezakhi zofuzo phantsi kolawulo lwabakhuthazi abathile. Umsebenzi we-beta-galactosidase olinganiswe kusetyenziswa i-ONPG unika ulwazi malunga nenqanaba lokubonakaliswa kwezakhi zofuzo.

Ukuhlolwa komsebenzi we-β-galactosidase: I-ONPG ingasetyenziswa njengendlela yokuhlola i-colorimetric kwitekhnoloji ye-DNA ephinda-phindayo ukuchonga ubukho kunye nokusebenza kwejini ye-LacZ, ebhala i-β-galactosidase. Le ndlela yokuhlola inceda ekuchongeni ii-clones eziqulethe ijini enomdla.

Izifundo ze-enzyme kinetics: I-ONPG ikwaluncedo ekufundeni i-kinetics ye-enzyme ye-β-galactosidase. Ngokulinganisa izinga le-enzyme-substrate reaction kwii-substrate concentrations ezahlukeneyo, kunokwenzeka ukumisela iiparameter ze-kinetic ezifana nee-Michaelis-Menten constants (Km) kunye ne-maximum reaction rates (Vmax).