Iimveliso

I-Monogalactopyranosides ye-fluorescein kunye ne-fluorescein methyl ester: i-synthesis, i-enzymatic hydrolysis nge-biotnylated β-galactosidase, kunye nokumiselwa kwe-coefficient yokuguqulwa kwe-diffusion.

I-Fluorescein monoglycosides (d-galactopyranoside (FMG) kunye ne-d-glucopyranoside) kunye ne-methyl ester (MFMG) yazo zilungiswe ukusuka kwi-acetobromoglucose / galactose kunye ne-fluorescein methyl ester kwizivuno ezilungileyo.Uvavanyo lwe-enzymatic hydrolysis (usebenzisa i-biotinylated β-galactosidase) ye-galacto derivatives yenziwe kwaye i-kinetic parameters ibalwe.Ukunyuka kwamaxesha e-15-20 ye-fluorescence intensity iye yabonwa ngexesha le-hydrolysis.Ukunyuka komgca we-fluorescence kuye kwaphawulwa ngexesha elifutshane kunye nokuxinwa okuphantsi kwe-substrate, okwenza ezi zixhobo zibe luncedo kunye neeprobes ezibuthathaka ze-galactosidases.Ubukhulu bexabiso leMichaelis–Menten (Km) leMFMG liphezulu kunolo lwe-FMG lucebisa ukuba kubekho utshintsho olunokubakho lwe-fluorogenic substrate.Ixabiso le-Km le-biotinylated β-Gal ene-FMG lingaphantsi kunelo le-enzyme yendalo.Olu qwalaselo lubonisa ubudlelwane obuphezulu be-substrate ye-enzyme ye-biotinylated xa kuthelekiswa ne-enzyme yendalo.I-Coefficients yokusasazwa koguqulelo iye yalinganiswa, kuzo zombini ii-substrates ze-fluorogenic kunye neemveliso zombini, zisebenzisa i-fluorescence correlation spectroscopy.Ii-coefficients zokuguqulelwa kokusasazwa kwee-fluorogenic substrates kunye neemveliso ze-enzymatic hydrolysis ziye zalinganiswa ukuba ziyafana, kuluhlu lwe-3.5-4.5 × 10−10 m2 s-1.Ngaloo ndlela ukongezwa okanye ukucuthwa kwe-enzymatic kinetics ngenxa yomahluko wokuguquguquka kwe-substrate kunye nemveliso ayibonakali kangako.