Kayayyaki

LecA (PA-IL) lectin cytotoxic ne da adhesin wanda Pseudomonas aeruginosa ya samar wanda ke ɗaure galactosides na hydrophobic tare da ƙayyadaddun ƙayyadaddun ƙayyadaddun alaƙa da alaƙa.Ta amfani da haɗin fassarar lecA-egfp da bincike na immunoblot na matrix na extracellular biofilm, muna nuna cewa an bayyana lecA a cikin sel masu girma na biofilm.A cikin gwaje-gwajen biofilm a tsaye akan duka polystyrene da bakin karfe, zurfin biofilm da ɗaukar hoto ya ragu ta hanyar maye gurbin lecA kuma an inganta shi a cikin LecA-overproducing iri PAO-P47.Halin yanayin yanayin biofilm ta mahaifar mahaifa, PAO-P47 amma ba mutancin lecA akan takaddun karfe ba shima an hana shi girma a gaban ko dai isopropyl-beta-D-thiogalactoside (IPTG) ko p-nitrophenyl-alpha-D-galactoside ( NPG).Bugu da ƙari, balagaggen nau'in halittun halittun daji da aka kafa idan babu waɗannan galactosides hydrophobic za a iya tarwatsa su ta hanyar ƙara IPTG.Sabanin haka, ƙari na p-nitrophenyl-alpha-L-fucose (NPF) wanda ke da alaƙa mai girma ga P. aeruginosa LecB (PA-IIL) lecti n ba shi da wani tasiri akan samuwar biofilm ko watsawa.Planktonic girma na P. aeruginosa PAO1 ba shi da tasiri ta kasancewar IPTG, NPG ko NPF, kuma nau'in ba zai iya amfani da waɗannan sugars a matsayin tushen carbon ba, yana nuna cewa tasirin da aka gani akan samuwar biofilm ya kasance saboda gasa na hana LecA-ligand. ɗaure.An kuma sami irin wannan sakamako don biofilms girma a ƙarƙashin yanayin kwarara mai ƙarfi akan takaddun karfe, yana nuna cewa LecA yana ba da gudummawa ga gine-ginen biofilm na P. aeruginosa a ƙarƙashin yanayin muhalli daban-daban.

An gano wani alpha-L-fucosidase (EC 3.2.1.51) wanda zai iya sakin ragowar t-fucosyl daga sashin gefe na xyloglucan oligosaccharides a cikin ganyen tsire-tsire na Arabidopsis.Haka kuma, an tsarkake wani alpha-L-fucosidase mai irin wannan ƙayyadaddun ƙayyadaddun kayan masarufi daga ganyen kabeji (Brassica oleracea) don ba da ƙungiya ɗaya akan SDS-PAGE.An samo jerin peptide guda biyu daga wannan rukunin furotin, kuma an yi amfani da su don gano lambar ƙirar Arabidopsis don alpha-fucosidase wanda muka ba da shawarar a kira AtFXG1.Bugu da kari, an kuma sami kwayar halittar Arabidopsis tare da homology tare da sanannun alpha-L-fucosidases, kuma mun ba da shawarar sanya masa suna AtFUC1.Dukansu AtFXG1 da ATFUC1 an bayyana su da yawa a cikin sel pastoris Pichia da ayyukan alpha-L-fucosidase da aka ɓoye ga matsakaicin al'ada.Alpha-L-fucosidase wanda AtFXG1 ya tsara yana aiki akan oligosaccharides daga xyloglucan XXFG da kuma 2'-fucosyl-lactitol amma ba akan p-nitrophenyl-alpha-L-fucopyranoside ba.Koyaya, AtFUC1 heterologously bayyana yana aiki ne kawai akan 2'-fucosyl-lactitol.Don haka, tsohon dole ne ya kasance yana da alaƙa da metabolism na xyloglucan.