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OALib Journal期刊
ISSN: 2333-9721
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瑞氏木霉内切葡聚糖酶ii在大肠杆菌中的重组表达及重组酶性质测定
, PP. 92-98
Keywords: 羧甲基纤维素钠盐,内切葡聚糖酶,大肠杆菌,β葡聚糖,瑞氏木霉
Abstract:
将不含信号肽的瑞氏木霉内切葡聚糖酶ii(cel5a)的cdna克隆到pet-28a(+)表达质粒上,与其n末端6个组氨酸标签序列融合,构建成pet-28a(+)-egl2表达质粒,在大肠杆菌bl21(de3)中诱导表达.利用低温诱导策略,成功表达出活性重组蛋白.westernblot结果显示重组cel5a相对分子分量大约为43000.进一步对重组cel5a进行ni-nta亲和层析柱纯化并对其进行酶学性质测定,结果显示以羧甲基纤维素钠为作用底物时重组酶最适作用ph为4.5,最适作用温度为60℃;重组酶热稳定性较好,在65℃及以下保温1.5h,活性仍相当稳定.mn2+对cel5a的酶活有促进作用,fe3+和cu2+有抑制作用,其它金属离子和edta对酶活没有明显影响.底物特异性研究表明,重组cel5a只能水解混合键葡聚糖和羧甲基纤维素钠,对混合键葡聚糖的水解能力是其对羧甲基纤维素钠水解能力的6.7倍,但对微晶纤维素、glassmicrofiberfilters、木聚糖、阿拉伯木聚糖和木葡聚糖没有水解作用.
| [1] | zhangyh,himmeme,mielenzjr.outlookforcellulaseimprovement:screeningandselectionstrategies[j].biotechn-
|
| [2] | penttileme,andrel,saloheimom,eta.lexpressionoftwotrichodermareeseiendoglucanasesintheyeastsaccharomyces
|
| [3] | shoemakers,schweickartv,ladnerm,eta.lmolecularcloningofexo-cellobiohydrolaseiderivedfromtrichodermareesei
|
| [4] | teeritt,salovuorii,knowlesj.themolecularcloningofthemajorcellulasegenefromtrichodermareesei[j].bio/tech-
|
| [5] | nology1,1983:696-699.
|
| [6] | ular-massendoglucanasefromtrichodermareeseiqm9414[j].applenvironmicrobio,l1998,64:555-563.
|
| [7] | saloheimom,nakari‘setalat,tenkanenm,eta.lcdnacloningofatrichodermareeseicellulaseanddemonstrationofen-
|
| [8] | doglucanaseactivitybyexpressioninyeast[j].eurjbiochem,1997,249:584-591.
|
| [9] | saloheimoa,henrissatb,hoffr-nam,eta.lanove,lsmallendoglucanasegene,egl5,fromtrichodermareeseiisolated
|
| [10] | byexpressioninyeast[j].molmicrobio,l1994,13:219-228.
|
| [11] | bypretreatmentwithfungalpectinasesandegtainvitro[j].plantphysiology,2008,147:1874-1885.
|
| [12] | sandhudk,kalramk.productionofcellulase,xylanaseandpectinasebytrichodermalongibrachiatumondifferentsub-
|
| [13] | no,l1998,16(2):54-60.
|
| [14] | okadah,sekiyat,yokoyamak,eta.lefficientsecretionoftrichodermareeseicellobiohydrolaseiiinschizosaccharomy-
|
| [15] | cespombeandcharacterizationofitsproducts[j].applmicrobiolbiotechno,l1998,49:301-308.
|
| [16] | oryzaeandtheirenzymaticproperties[j].jbiotechno,l1998,65:163-171.
|
| [17] | macarronr,acebalc,castillonmp,eta.lmodeofactionofendoglucanaseiiifromtrichodermareesei[j].biochemj,
|
| [18] | 1993,289:867-873.
|
| [19] | oladv,2006,24:452-481.
|
| [20] | cerevisiae[j].yeast,1987,3:175-185.
|
| [21] | strainl27[j].bio/technology1,1983:691-696.
|
| [22] | penttil-m,lehtovaarap,nevalainenh,eta.lhomologybetweencellulasegenesoftrichodermareesei:completenucleotide
|
| [23] | sequenceoftheendoglucanaseigene[j].gene,1986,45:253-263.
|
| [24] | saloheimom,lehtovaarap,penttil-m,eta.legiii,anewendoglucanasefromtrichodermareesei:thecharacterizationof
|
| [25] | bothgeneandenzyme[j].gene,1988,63(1):11-22.
|
| [26] | okadah,tadak,sekiyat,eta.lmolecularcharacterizationandheterologousexpressionofthegeneencodingalow-molec-
|
| [27] | barnettcc,berkarm,fowlert.cloningandamplificationofthegeneencodinganextracellularβglucosidasefromtri-
|
| [28] | chodermareesei:evidenceforimprovedratesofsaccharificationofcellulosicsubstrates[j].biotechnology(ny),1991,9:
|
| [29] | 562-567.
|
| [30] | wangt,liuxm,yuq,eta.ldirectedevolutionforengineeringphprofileofendoglucanaseiiifromtrichodermareesei
|
| [31] | expressedinsaccharomycescerevisiaewithhigherglycosylationandstability[j].proteinexpresspuri,f2008,58:162-167.
|
| [32] | zhaoqx,yuans,wangx,eta.lrestorationofmatureetiolatedcucumberhypocotylcellwallsusceptibilitytoexpansin
|
| [33] | strates[j].transbrmycolsoc,1982:409-413.
|
| [34] | logemannj,schellj,willmitzerl.improvedmethodfortheisolationofrnafromplant-tissues[j].analbiochem,
|
| [35] | 1987,163(1):16-20.protein-dyebinding[j].analbiochem,1976,72:248-254.
|
| [36] | hanningg,makridessc.strategiesforoptimizingheterologousproteinexpressioninescherichiacoli[j].trendsbiotech-
|
| [37] | takashimas,iikurah,nakamuraa,eta.loverproductionofrecombinanttrichodermareeseicellulasesbyaspergillus
|
| [38] | [j].biomoleng,2005,22:89-94.
|
| [39] | qinyq,weixm,liuxm,eta.lpurificationandcharacterizationofrecombinantendoglucanaseoftrichodermareesei
|
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