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OALib Journal期刊
ISSN: 2333-9721
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木聚糖酶xynb分子ser/thr平面导入精氨酸对酶热稳定性的影响
DOI: 10.3969/j.issn.1000-2006.2014.04.020, PP. 107-112
Keywords: 定点突变,木聚糖酶,精氨酸,热稳定性
Abstract:
为提高木聚糖酶的热稳定性,通过定点突变的方法,在来源于黑曲霉aspergillusnigernl-1的木聚糖酶xynb分子ser/thr平面上引入5个精氨酸,实现了突变酶在毕赤酵母中的表达。通过酶学性质的研究表明,突变酶xynb-104和xynb-77的最适反应温度为50℃,且与原酶相比相对酶活力得到显著提高。在存在底物的情况下,50℃热处理2h,突变酶xynb-104和xynb-77残余酶活力较原酶的相对转化效率由26%提高到80%左右。突变酶xynb-104的最适反应ph与原酶一致,而突变酶xynb-77最适ph由5.0提高到5.5。结果表明,在木聚糖酶分子ser/thr平面不同折叠片引入精氨酸可以增强酶结构的稳定性,特别是能显著提高热稳定性。
| [1] | amalias,johanw,christophel,etal.theendoxylanasesfromfamily11:computeranalysisofproteinsequencesrevealsimportantstructuralandphylogeneticrelationships[j].journalofbiotechnology,2002,95(2):109-131.
|
| [2] | qiuzh,shipj,luohy,etal.axylanasewithbroadphandtemperatureadaptabilityfromstreptomycesmegasporusdsm41476,anditspotentialapplicationinbrewingindustry[j].enzymeandmicrobialtechnology,2010,46(6):506-512.
|
| [3] | khandeparkerr,numanmt.bifunctionalxylanasesandtheirpotentialuseinbiotechnology[j].journalofindustrialmicrobiology&biotechnology,2008,35(7):635-644.
|
| [4] | 杨浩萌,柏映国,李江,等.木聚糖酶xynb的n46d突变、表达及酶学性质变化[j].中国生物化学与分子生物学报,2006,22(3):204-211.yanghm,baiyg,lij,etal.n46dmutagenesisofxylanasexynbandenzymaticchangesofexpressionproduct[j].chinesejournalofbiochemistryandmolecularbiology,2006,22(3):204-211.
|
| [5] | turuneno,vuoriom,fenelf,etal.engineeringofmultipleargininesintotheser/thrsurfaceoftrichodermareeseiendo-1,4-beta-xylanaseiiincreasesthethermotoleranceandshiftsthephoptimumtowardsalkalineph[j].proteinengineering,2002,15(2):141-145.
|
| [6] | 周晨妍.宇佐美曲霉木聚糖酶基因的克隆、表达及定向进化研究[d].无锡:江南大学,2008.zhoucy.cloning,expressionanddirectedmutagenesisofxylanasegenefromaspergillususamii[d].wuxi:jiangnanuniversity,2008.
|
| [7] | lif,yangsy,zhaolg,etal.synonymouscondonusagebiasandoverexpressionofasyntheticxynbgenefromaspergillusnigernl-1inpichiapastoris[j].bioresource,2012,7(2):2330-2343.
|
| [8] | millergl.useofdinitrosalicylicacidreagentfordeterminationofreducingsugar[j].analyticalchemistry,1959,31(3):426-428.
|
| [9] | liumq,liugf.expressionofrecombinantbacilluslicheniformisxylanaseainpichiapastorisandxylooligosaccharidesreleasedfromxylansit[j].proteinexpressionandpurification,2008,57(2):101-107.
|
| [10] | fredf,mattil,jannej,etal.adenovodesignedn-terminaldisulphidebridgestabilizesthetrichodermareeseiendo-1,4-xylanaseii[j].journalofbiotechnology,2004,108(2):137-143.
|
| [11] | taox,qinw.directedevolutionofstreptomyceslividansxylanasebtowardenhancedthermalandalkalinephstability[j].worldjournalofmicrobiologyandbiotechnology,2009,25(1):93-100.
|
| [12] | chenxz,xusq,zhums,etal.site-directedmutagenesisofanaspergillusnigerxylanasebanditsexpression,purificationandenzymaticcharacterizationinpichiapastoris[j].processbiochemistry,2010,45(1):75-80.
|
| [13] | lif,zhaolg.,ligq,etal.cloning,expressionandcharacterizationofxylanasegenexynbfromaspergillusnigerinpichiapastoris[c]//sunruncang,fushiyu.4thisetpp.guangzhou:southchinauniversityoftechnologypress,2010:637-640.
|
| [14] | sriprangr,asanok,gobsukj,etal.improvementofthermostabilityoffungalxylanasebyusingsite-directedmutagenesis[j].journalofbiotechnology,2006,126(4):454-462.
|
| [15] | 杨浩萌,姚斌,罗会颖,等.木聚糖酶xynb分子中折叠股b1和b2间的疏水作用对酶热稳定性的影响[j].生物工程学报,2005,21(3):414-419.yanghm,yaob,luohy,etal.hydrophobicinteractionbetweenβ-sheetb1andb2inxylanasexynbinfluencingtheenzymethermostability[j].chinesejournalofbiotechnology,2005,21(3):414-419.
|
| [16] | kimsh,pokhrels,yooyj.mutationofnon-conservedaminoacidssurroundingcatalyticsitetoshiftphoptimumofbacilluscirculansxylanase[j].journalmolecularcatalysisb:enzymatic,2008,55(3):130-136.
|
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