Cov khoom

Monogalactopyranosides ntawm fluorescein thiab fluorescein methyl ester: synthesis, enzymatic hydrolysis los ntawm biotnylated β-galactosidase, thiab kev txiav txim siab ntawm translational diffusion coefficient

Fluorescein monoglycosides (d-galactopyranoside (FMG) thiab d-glucopyranoside) thiab lawv cov methyl ester (MFMG) tau npaj los ntawm acetobromoglucose / galactose thiab fluorescein methyl ester hauv cov txiaj ntsig zoo.Enzymatic hydrolysis thwmsim (siv biotinylated β-galactosidase) ntawm galacto derivatives tau ua thiab kinetic parameters raug xam.Ib qho 15-20 zaug nce ntawm cov fluorescence siv tau raug pom thaum lub sij hawm hydrolysis.Ib qho linear nce ntawm fluorescence tau raug sau tseg nyob rau lub sij hawm luv luv thiab tsis tshua muaj concentration ntawm substrate, ua rau cov tebchaw no pab tau thiab rhiab soj ntsuam rau galactosidases.Qhov loj ntawm Michaelis-Menten tas li (Km) tus nqi rau MFMG yog siab dua li ntawm FMG qhia txog qhov muaj peev xwm hloov pauv ntawm fluorogenic substrate.Km tus nqi rau biotinylated β-Gal nrog FMG yog qis dua qhov ntawd rau cov enzyme ib txwm muaj.Qhov kev soj ntsuam no qhia tau hais tias ntau dua substrate affinity ntawm biotinylated enzyme nyob rau hauv kev sib piv rau cov haiv neeg enzyme.Kev txhais lus diffusion coefficients tau ntsuas, rau ob qho tib si fluorogenic substrates thiab ob qho tib si cov khoom, siv fluorescence correlation spectroscopy.Translational diffusion coefficients rau fluorogenic substrates thiab cov khoom enzymatic hydrolysis tau raug ntsuas kom zoo ib yam, nyob rau hauv thaj tsam ntawm 3.5–4.5 × 10–10 m2 s−1.Yog li ib qho kev txhim kho lossis retardation ntawm cov enzymatic kinetics vim qhov sib txawv ntawm kev hloov pauv hloov pauv ntawm substrate thiab cov khoom lag luam tsis pom meej.