Imikhiqizo

I-LecA (PA-IL) iyi-lectin eyi-cytotoxic kanye ne-adhesin ekhiqizwa i-Pseudomonas aeruginosa ehlanganisa i-hydrophobic galactosides ngokucaciswa okuphezulu nokuhambisana.Ngokusebenzisa ukuhlanganiswa kokuhumusha kwe-lecA-egfp kanye nokuhlaziywa kwe-immunoblot ye-biofilm extracellular matrix, sibonisa ukuthi i-lecA iboniswa kumaseli akhuliswe yi-biofilm.Ezivivinyweni ezimile ze-biofilm kukho kokubili i-polystyrene nensimbi engagqwali, ukujula kwe-biofilm nokumbozwa kwendawo kwehliswe ngokuguqulwa kwe-lecA futhi kwathuthukiswa kuhlobo olukhiqiza ngokweqile lwe-LecA i-PAO-P47.Ukumbozwa kwendawo ye-Biofilm wuhlobo lomzali, i-PAO-P47 kodwa hhayi i-lecA eguquguqukayo kumakhuphoni ensimbi futhi kwavinjwa ukukhula phambi kwe-isopropyl-beta-D-thiogalactoside (IPTG) noma i-p-nitrophenyl-alpha-D-galactoside ( NPG).Ngaphezu kwalokho, ama-biofilms ohlobo lwasendle avuthiwe akhiwe ngaphandle kwalawa ma-galactoside e-hydrophobic angahlakazwa ngokungezwa kwe-IPTG.Ngokuphambene, ukungezwa kwe-p-nitrophenyl-alpha-L-fucose (NPF) okunokuhambisana okuphezulu kwe-P. aeruginosa LecB (PA-IIL) lecti n akuzange kube nomthelela ekwakhiweni kwe-biofilm noma ekuhlakazweni.Ukukhula kwe-Planktonic ye-P. aeruginosa PAO1 akuthintwanga ukuba khona kwe-IPTG, NPG noma i-NPF, futhi uhlobo loshukela alukwazanga ukusebenzisa laba shukela njengemithombo yekhabhoni, okuphakamisa ukuthi imiphumela ebonwe ekwakhiweni kwe-biofilm yayingenxa yokuvinjwa kokuncintisana kwe-LecA-ligand. ukubopha.Imiphumela efanayo iphinde yatholwa kuma-biofilms atshalwa ngaphansi kwezimo zokugeleza okuguquguqukayo kumakhuphoni ensimbi, okuphakamisa ukuthi i-LecA inikela ekwakhiweni kwe-P. aeruginosa biofilm ngaphansi kwezimo zemvelo ezihlukene.

I-alpha-L-fucosidase (EC 3.2.1.51) ekwazi ukukhulula izinsalela ze-t-fucosyl kusukela kuchungechunge oluseceleni lwe-xyloglucan oligosaccharides iye yatholwa emaqabunga ezitshalo ze-Arabidopsis.Ngaphezu kwalokho, i-alpha-L-fucosidase enezici ezifanayo ze-substrate yahlanzwa emaqabunga eklabishi (Brassica oleracea) ukuze inikeze ibhendi eyodwa ku-SDS-PAGE.Ukulandelana kwe-peptide okubili kutholwe kuleli bhendi yephrotheni, futhi kwasetshenziselwa ukukhomba ikhodi yofuzo ye-Arabidopsis ye-alpha-fucosidase esiphakamisa ukuyibiza i-AtFXG1.Ngaphezu kwalokho, isakhi sofuzo se-Arabidopsis esine-homology ene-alpha-L-fucosidase eyaziwayo nayo itholakele, futhi sihlongoze ukuyiqamba ngokuthi i-AtFUC1.Kokubili i-AtFXG1 ne-ATFUC1 zivezwe ngendlela exakile kumaseli e-Pachia pastoris kanye nemisebenzi ye-alpha-L-fucosidase efihliwe endaweni yamasiko.I-alpha-L-fucosidase efakwe ikhodi yi-AtFXG1 ibisebenza ngokumelene nama-oligosaccharides asuka ku-xyloglucan XXFG kanye ne-2'-fucosyl-lactitol kodwa hhayi ngokumelene ne-p-nitrophenyl-alpha-L-fucopyranoside.Nokho, i-AtFUC1 evezwe nge-heterologously yayisebenza kuphela ngokumelene ne-2'-fucosyl-lactitol.Ngakho-ke, okokuqala kufanele kuhlobane ne-xyloglucan metabolism.