Zogulitsa

Monogalactopyranosides wa fluorescein ndi fluorescein methyl ester: kaphatikizidwe, enzymatic hydrolysis ndi biotnylated β-galactosidase, ndi kutsimikiza kwa matanthauzo a diffusion coefficient.

Fluorescein monoglycosides (d-galactopyranoside (FMG) ndi d-glucopyranoside) ndi methyl ester (MFMG) yawo yakonzedwa kuchokera ku acetobromoglucose/galactose ndi fluorescein methyl ester pobereka bwino.Zoyeserera za enzymatic hydrolysis (pogwiritsa ntchito biotinylated β-galactosidase) zochokera ku galacto zachitika ndipo magawo a kinetic adawerengedwa.Kuwonjezeka kwa 15-20 kwa mphamvu ya fluorescence kwawonedwa panthawi ya hydrolysis.Kuwonjezeka kwa mzere wa fluorescence kwadziwika pakanthawi kochepa komanso kutsika kwa gawo lapansi, zomwe zimapangitsa kuti mankhwalawa akhale othandiza komanso ozindikira ma galactosidase.Ukulu wa mtengo wa Michaelis–Menten (Km) wa MFMG ndi wapamwamba kuposa wa FMG zomwe zikuwonetsa kusintha kosinthika kwa gawo lapansi la fluorogenic.Mtengo wa Km wa biotinylated β-Gal wokhala ndi FMG ndi wocheperako kuposa wa enzyme yakubadwa.Kuwona uku kukuwonetsa kuyanjana kwapang'onopang'ono kwa enzyme ya biotinylated poyerekeza ndi enzyme yakubadwa.Ma coefficients omasuliridwa ayesedwa, pamagawo onse a fluorogenic ndi zinthu zonse, pogwiritsa ntchito mawonekedwe a fluorescence correlation spectroscopy.Ma coefficients omasuliridwa a ma substrates a fluorogenic ndi zinthu za enzymatic hydrolysis adayesedwa kukhala ofanana, mumitundu ya 3.5-4.5 × 10−10 m2 s−1.Chifukwa chake kukulitsa kapena kuchedwetsa kwa enzymatic kinetics chifukwa cha kusiyana kwa kumasulira kwa gawo lapansi ndi zinthu sizikuwonekera.