Paecilomyces sp.FLH30内切β1 34葡聚糖酶基因和Selenomonas ruminantium木糖苷酶基因在赤酵母中的表达.docx

Expression Expression of the Paecilomyces sp．FLH 30 endo p-1，3(4) Glucanase Gene and Selenomonas ruminantium Xylosidase gene in Pich ia pastoris Abstract From Pecilomyces sp．FLH 30 13-1，3(4)一glucanase has an effect on the cell wall of plants，it could efficiently hydrolysis phm cell wallSeleno聊nas ruminantium is a dominant bacteria in the rumen of the rumen,The production of Selenomonas ruminantm plays a key role in the process of complete degradation of xylanase．111is exper硫nt takes Paecilomyces sp．FLH30 endo 13-1，3(4)glucanase gene and Selenomonasruminantium xylosidase gene as the research object．ne results are as folbws： Optimization ofcodon sequence ofPaecilomyces sp．FLH30 endo 13-l，3(4)glucanase，build new carrier，using Sac I enzyme digestion DNA，by 1 500V of the current stimulation,transduction into the strain GS 1 1 5，and the level of test tube，shake flask and fermentation tank were expressed respectively， the enzyme activity increased from the original 11．26 II『／mI，to 259．73 IU／mI，．，nle results of recombinam enzyme GLUnm measured in the exlDerimem showed：ne change of temperature in the temperature of 37．50。C is small，and the change is not remarkable证the category of pH 5．O一8．0． GLUnm could hydrolySiS of Lichenim and Barley 13-ghcan,it has hydrolysis effect on Oat spelt xylan,CMC-Na and pNPX． Optimize the synthesis of xylos idase gene，get a new xylos idase gene 3SAnm,construction of recombinant vector pPIC9K-XSAnm,。uS崦题，II enzyme digestion DNA，by 1500 V of the current stimulation，transduction into the strain GSl 15，It is expressed in three different expression environments，in the test tube，in shake flask,in 10 L fermentation tank to express，the recombinam enzyme activ毋was increased from 17．54 IU／mL to 287．61 IU／mL．m enzymatic property experimems showed：T】k recombinant xylosidase enzyme was stable ha pH 5．0—6．0 and the temperature was 40—55℃，The recombinam xylos idase can degrade pNPX,Oat spelt xylan，the recombinam xylosidase had no hydro舾is effect on CMC—Na，L