Nā huahana

Monogalactopyranosides o ka fluorescein a me ka fluorescein methyl ester: synthesis, enzymatic hydrolysis e biotnylated β-galactosidase, a me ka hoʻoholo ʻana o ka unuhi diffusion coefficient

Ua hoʻomākaukau ʻia nā monoglycosides Fluorescein (d-galactopyranoside (FMG) a me d-glucopyranoside) a me kā lākou methyl ester (MFMG) mai acetobromoglucose/galactose a me fluorescein methyl ester i nā hua maikaʻi.Ua hana ʻia nā hoʻokolohua hydrolysis enzymatic (e hoʻohana ana i ka biotinylated β-galactosidase) o nā derivatives galacto a ua helu ʻia nā ʻāpana kinetic.Ua ʻike ʻia ka piʻi ʻana o ka fluorescence he 15-20 manawa i ka wā o ka hydrolysis.Ua ʻike ʻia ka piʻi ʻana o ka fluorescence i ka manawa pōkole a me ka haʻahaʻa haʻahaʻa o ka substrate, e hoʻohana ana i kēia mau pūhui i mea pono a naʻauao no nā galactosidases.ʻOi aku ka nui o ka waiwai mau Michaelis–Menten (Km) no MFMG ma mua o ka FMG e hōʻike ana i kahi hoʻololi conformational hiki ke hoʻololi i ka substrate fluorogenic.ʻOi aku ka haʻahaʻa o ka helu Km no ka β-Gal biotinylated me FMG ma mua o kēlā no ka enzyme maoli.Hōʻike kēia ʻike i ka pili ʻana o ka substrate kiʻekiʻe o ka enzyme biotinylated i hoʻohālikelike ʻia me ka enzyme maoli.Ua ana ʻia nā ʻāpana diffusion unuhi, no nā substrates fluorogenic a me nā huahana ʻelua, e hoʻohana ana i ka spectroscopy correlation fluorescence.Ua ana ʻia nā mea hoʻoheheʻe unuhi no nā substrate fluorogenic a me nā huahana hydrolysis enzymatic e like, ma ka laulā o 3.5–4.5 × 10−10 m2 s−1.No laila, ʻaʻole i ʻike ʻia ka hoʻonui a i ʻole ka hoʻihoʻi ʻana o nā kinetics enzymatic ma muli o ka ʻokoʻa o ka neʻe ʻana o ka substrate a me ka huahana.