Yimaphi amazinga okushisa alungele umsebenzi we-benzonase?

Ukubuka konkeIBenzonaseUkusebenza kwe-enzymatic

IBenzonase iyinzyme eguquguqukayo edlala indima ebalulekile emkhakheni we-biotechnology, ikakhulukazi ekususweni kwama-acid ama-nucleic kusuka kumasampula endalo. Njenge-non endonuclease ethize ethathwe kuyoISerratia Marscens, Benzinase unamathela ngempumelelo zombili ama-DNA nama-rolekyuli e-rna. Lesi sici sikwenza kube nzima kakhulu kuzicelo ezahlukahlukene, njengokuhlanza amaprotheni aphindaphindwayo kanye nokuqinisekisa ubuqotho bezinqubo zokukhiqiza zokugoma. Abakhiqizi kanye nabaphakeli be-biochemical reagents bavame ukugcizelela ukusebenza kahle kweBenzonase kuma-nucleic acid angonakaliso ngaphandle kokulandelana - Izidingo ezithile, okuholela ekukhuthaleni kwayo -

Indlela yokwenza isenzo

I-Benzonase isebenza nge-hydrolyzing nge-phospolyzing ama-phosphate dider bond kuma-nucleic acid, akhiqiza ama-oligonucleotides ayi-5 ' I-enzyme isebenza kakhulu ngokumelene ne-single - ehlelwe futhi iphindwe kabili - ama-nucleic acid, kungaba aqondile, isiyingi, noma i-supercoiled. Leli khono lokunciphisa ukungcoliswa kwe-nucleic acid liyindlela ebalulekile ezinqubweni lapho izinsalela ze-nucleic acid zingaphazamisa izinhlelo zokusebenza ezansi.

Izicelo ze-benzonase e-biotechnology

Embonini ye-biotechnology, iBenzonase isetshenziswa kakhulu ngamakhono ayo we-nucleic acid acidi. Izicelo zalo zichithe izizinda ezahlukahlukene, kufaka phakathi ukukhiqizwa kokugoma, lapho ukususwa kwamaseli we-Hot Cell Nucleic Acid kubalulekile ekuphepheni nasekusebenzeni kahle komkhiqizo wokugcina.

Indima Emkhiqizweni Wokugomela

Imboni yezemithi ithembele eBenzonase ngobungcweti bayo ekunciphiseni ukungcoliswa kwe-nucleic acid kumazinga we-picogram. Lokhu kuqinisekisa ukuthi kugoma kanye namanye amaprotheni - Ama-The Lacheditics asekelwe kugcina amazinga aphezulu wokuhlanzeka. Abakhiqizi babeka phambili iBenzonase ezinhlangeni zabo zokukhiqiza ukuze bahlangabezane nezindinganiso eziqinile zokulawula.

Ukuphuculwa kwezinqubo zokuhlunga

I-Benzonase ayigcini nje ngokususa ukususwa kwe-nucleic acid kodwa futhi nengculazi ekunciphiseni ukubonwa kwe-cell Lysates. Lokhu kuncishiswa kwe-viscosity kukhuthaza ukugcwaliswa okusebenzayo kanye nezinqubo ze-ultrafiltration, okuholele ekuvuseleleni ngcono kanye nobumsulwa bemikhiqizo yemvelo. Abahlinzeki bemishini ye-bioprocessing bavame ukuncoma i-benzonase ukwenza ngcono ukugeleza komsebenzi wokuhlanzwa.

Ukuqonda izinga lokushisa elifanele lomsebenzi weBenzonase

Ukushisa kudlala indima ebalulekile ekusebenzeni nasekuzinzweni kwama-enzyme, kufaka neBenzonase. Ukuhlonza izinga lokushisa elifanele lomsebenzi walo lingathonya kakhulu ukusebenza kwezindlela ezisetshenziswe kuzo.

I-Enzyme Kinetics kanye lokushisa

I-Enzyme Kinetics iyazwela ukuhlukahluka kwezinga lokushisa, ngomsebenzi ovame ukukhuphuka ngamazinga okushisa akhuphukayo kuze kube yilapho inani eliphakeme lifinyelelwe. Ngale ndlela yokushisa efanelekile, umsebenzi we-enzyme wehla ngokushesha ngenxa yokonakaliswa. OkweBenzonase, izinga lokushisa elifanele livame ukubhekwa njengezindawo ezingaba ngu-37 degrees Celsius, ezihambelana nezimo zomzimba.

Lokushisa - Umsebenzi othembele weBenzonase

Umsebenzi weBenzonase uthonywa ikakhulukazi ngamazinga okushisa, athinta isilinganiso sawo se-caslytic nokuqina. Ucwaningo lukhombisa ukuthi emazingeni okushisa aphansi, umsebenzi we-enzyme uncishisiwe, kanti emazingeni okushisa aphezulu, ubungozi bokukhuphuka kwe-denaturation.

Ubufakazi Bokuhlola

Ezimweni zelebhu, amaBenzonase ekhombisa umsebenzi omkhulu cishe ngama-degree angama-37. Lokhu kushisa kuvumela ukuqina kwe-nucleic acid cleavage ngenkathi kugcinwa ukuqina kwe-enzyme. Amafektri abhekelela ekukhiqizweni kwe-enzyme kuvame ukulinganisa izinqubo zawo ukuze kuxhaswe la mazinga okushisa, aqinisekise ukusebenza kwe-enzyme okungaguquki.

Indima Yokushisa ku-Enzyme Evition

Ngenkathi amazinga okushisa aphezulu athuthukisa umsebenzi we-enzyme, angathonya futhi ukuba semaphukule ku-inhibitors. Ukugcina izinga lokushisa elifanele kubalulekile ekunciphiseni imiphumela yokuvinjwa engahle iphinde iphindekenze umsebenzi weBenzonase.

Inhibitor ukuzwela

Ukuba khona kwama-reagents athile kungavimbela umsebenzi we-benzonase kuze kufike ku-50% ekugxileni okuthile. Kodwa-ke, ukulondolozwa kwe-enzyme ekushiseni kwayo okuhle kunganciphisa le miphumela, kuqinisekiswe ukusebenza okwengeziwe ezisetshenziselwa izimboni.

Ukucatshangelwa okungokoqobo kokulawulwa kwezinga lokushisa ezinhlelweni zezimboni

Ukulawulwa kokushisa kubalulekile ukuze kwandise ukusebenza kahle kweBenzonase kwezimboni - Izinqubo ezikade. Bobabili abakhiqizi nabaphakeli badinga ukusebenzisa izinhlelo eziqinile eziqinisekisa izimo zokushisa ezingaguquki.

Ukuqapha okushisa kanye nokulungiswa

Ukuqapha njalo kwamazinga okushisa, kuhambisana nezinhlelo zokushintsha okuzenzakalelayo zokushisa, kungasiza ukugcina umsebenzi ofanele we-enzyme. Amafektri ajwayele ukutshala imali kubuchwepheshe bokubala lokushisa okuthuthukile ukuze aqinisekise ukuthi izinqubo zabo zokukhiqiza zihlala zisebenza kahle futhi zibizelisa - ngempumelelo.

Ukuzimisela kokuhlola kwezinga lokushisa eliphakeme

Ukunquma izinga lokushisa i-Optima le-benzonase kufaka phakathi ukuhlolwa okuhlelekile, okuvumela ukulinganiswa okuqondile kwezicelo zezimboni. Lokhu kuqinisekisa ukuthi amandla e-enzyme ahlanzeke ngokuphelele ezinqubweni ezahlukahlukene ze-biotechchological.

Izindlela Zokuhlola

Izivumelwano ezijwayelekile zokuhlola zibandakanya ukuhlola umsebenzi we-benzonase kuwo wonke amazinga okushisa. Abakhiqizi bavame ukuqhuba lokhu kuhlolwa ngesikhathi sesigaba sokuthuthukiswa kwe-enzyme ukusungula izimo zokushisa ezisebenzayo kakhulu zokuhanjiswa kwezicelo ezahlukahlukene.

Qhathanisa i-benzonase namanye ama-endonucleases

IBenzonase imele phakathi kwama-endonucleses ngenxa yomsebenzi wayo obanzi Izifundo zokuqhathanisa zisiza ukugcizelela lezi zinzuzo ezisetshenziselwa izicelo ezahlukahlukene.

Izinzuzo ngaphezu kwamanye ama-enzyme

Uma kuqhathaniswa namanye ama-endonucleases, i-benzonase inikeza ukucekelwa phansi okuphezulu kwe-nucleic acid kulo lonke uhla lokushisa olubanzi. Lokhu kuguquguquka kubalulekile ukuthi abakhiqizi bafuna izisombululo ezithembekile nezisebenzayo ze-enzymatic zezinselelo eziyinkimbinkimbi ze-bioproprocessing.

Izinkomba zokucwaninga zesikhathi esizayo ekusebenzeni kokushisa kwe-enzyme

Ucwaningo oluqhubekayo luyaqhubeka nokuhlaziya ukuqonda kwethu ukuqonda kwamandla okushisa kwe-enzyme, ngenhloso yokuthuthukisa ukusebenza kahle kweBenzonase kanye nokuqina. Lolu cwaningo lubalulekile ukuze kugcinwe umphetho we-enzyme emkhakheni wokuncintisana we-bioprocessing.

Izinto ezintsha ku-enzyme ubunjiniyela

Intuthuko e-Protein Engineering ingaholela ekuthuthukisweni kweBenzonase Variants ngokuqina kwezinga lokushisa okuthuthukile kanye nomsebenzi. Ngokugxila kulezi zinto ezintsha, abakhiqizi kanye nabahlinzeki banganikela ngezixazululo ezisebenzayo ze-enzymatic ezihambelana nezidingo ezithile zezimboni.

I-Bluekit ihlinzeka ngezixazululo

I-Bluekit inikeza izixazululo ezihambelana nezenzelwe ukwenza kahle umsebenzi weBenzonase kuzo zonke izimo zokushisa ezahlukahlukene. Izinhlelo zethu ze-bioprocessing ezithuthukisiwe ziqinisekisa ukulawulwa kokushisa okuqondile, ukuthuthukisa ukusebenza kahle kwe-enzyme nokuqina. Ngokuhlanganisa isimo - we - The - Ukuqapha ubuchwepheshe kanye nobuchwepheshe bokulungiswa, sisiza abakhiqizi kanye nabahlinzeki bagcina ikhwalithi yokukhiqiza engaguquki. Ubuchwepheshe bethu ezisoleni ze-enzymatic bukuhlomisa amathuluzi adingekayo ukubhekana nezinselelo eziyinkimbinkimbi ze-bioprocusting, uqinisekise ukusebenza kwe-enzyme kahle kanye nobumsulwa bomkhiqizo.