Ibicuruzwa

Monogalactopyranoside ya fluorescein na fluorescein methyl ester: synthesis, hydrolysis enzymatique na biotnylated β-galactosidase, no kugena coefficient de diffuzione yubusobanuro.

Fluorescein monoglycoside (d-galactopyranoside (FMG) na d-glucopyranoside) hamwe na methyl ester (MFMG) byateguwe kuva acetobromoglucose / galactose na fluorescein methyl ester mu musaruro mwiza.Ubushakashatsi bwa hydrolysis ya Enzymatique (ukoresheje biotinylated β-galactosidase) ikomoka kuri galacto yarakozwe kandi ibipimo bya kinetic byabazwe.Kwiyongera inshuro 15-20 kwiyongera kwa fluorescence byagaragaye mugihe cya hydrolysis.Kwiyongera kumurongo wa fluorescence byagaragaye mugihe gito hamwe nubushyuhe buke bwa substrate, bigatuma ibyo bikoresho bigira akamaro kandi byoroshye kuri galactosidase.Ubunini bwa Michaelis - Menten burigihe (Km) agaciro ka MFMG burenze ubw'ubwa FMG bwerekana ko impinduka zishobora guhinduka ziterwa na fluorogenic substrate.Km agaciro ka biotinylated β-Gal hamwe na FMG iri munsi yibyo kuri enzyme kavukire.Iri genzura ryerekana isano iri hejuru ya enzyme ya biotinylated ugereranije na enzyme kavukire.Ibipimo byo gukwirakwiza ibisobanuro byapimwe, kuri substrate ya fluorogenic n'ibicuruzwa byombi, ikoresha fluorescence ihuriro rya spekitroscopi.Coefficients yo gukwirakwiza ibisobanuro bya fluorogenic substrate hamwe nibicuruzwa bya hydrolysis ya enzymatique byapimwe bisa, murwego rwa 3.5-4.5 × 10−10 m2 s - 1.Rero, kuzamura cyangwa kudindiza kinetic enzymatique kubera itandukaniro ryimikorere yimikorere ya substrate nibicuruzwa ntabwo bigaragara.