|
OALib Journal期刊
ISSN: 2333-9721
费用：99美元
|
|
|
|
以类弹性蛋白多肽为标签纯化1,3-丙二醇氧化还原酶
DOI: 10.3724/SP.J.1145.2013.00709, PP. 709-712
Keywords: 类弹性蛋白多肽,纯化标签,可逆相变,重组酶纯化,非色谱法
Abstract:
以类弹性蛋白多肽（elps）为标签的非色谱纯化重组蛋白显示出良好的应用前景.本文使用自行设计的新型elps纯化标签，以低浓度盐诱发其可逆相变，从而更温和、高效地纯化融合表达的1,3-丙二醇氧化还原酶（pdor）.在测得elps-pdor融合蛋白在不同浓度na2co3溶液中相变温度的基础上，选取0.8mol/lna2co3溶液，在室温下使elps发生可逆相变，与目的蛋白pdor一同聚集，经可逆相变以离心法得以纯化，得到纯度约98%的纯酶，纯化倍数可达到8倍以上，是组氨酸标签的7倍多.融合酶的酶学性质表明elps作为纯化标签对目标蛋白空间结构影响微小，融合蛋白可直接作为酶催化化学反应.低盐诱发相变的方法，可避免盐浓度过高对酶结构及性能产生的不利影响，因此，在重组酶纯化上更具有优势.图3表2参17
| [1] | bankimr,fengl,wooddw.simplebioseparationsusingself-cleavingelastin-likepolypeptidetags[j].natmethods,2005,2:659-661
|
| [2] | baleyaf,wuwy,davidww.optimizationofelp-inteinmediatedproteinpurifcationbysaltsubstitution[j].proteinexpresspurif,2009,66:198-202
|
| [3] | chowd,nunaleeml,limdw,simnickaj,chilkotia.peptide-basedbiopolymersinbiomedicineandbiotechnology[j].matersciengrep,2008,62:125-155
|
| [4] | 黄凯宗,李晶晶,王文研,李巍,张光亚.类弹性蛋白多肽的从头设计、非色谱纯化及盐效应[j].生物工程学报,2011,27(4):661-666[huangkz,lijj,wangwy,liw,zhanggy.denovodesign,non-chromatographicpurificationandsaltseffectofelastin-likepolypeptides[j].chinjbiotech,2011,27(4):661-
|
| [5] | meyerde,chilkotia.geneticallyencodedsynthesisofprotein-basedpolymerswithpreciselyspecifiedmolecularweightandsequencebyrecursivedirectionalligation:examplesfromtheelastin-likepolypeptidesystem[j].biomacromolecules,2002,3:357-367
|
| [6] | 付晓平,王文研,张光亚.以类弹性蛋白多肽为标签的表达质粒构建及其用于木聚糖酶的非色谱纯化[j].微生物学报,2012,52(1):90-95[fuxp,wangwy,zhanggy.constructionofanexpressionvectorwithelastin-likepolypeptidetagtopurifyxylanase[j].actamicrobiolsin,2012,52(1):90-
|
| [7] | 方慧英,张成,诸葛斌,诸葛健.产1,3-丙二醇新型基因工程菌的构建[j].应用与环境生物学报,2009,15(5):708-712[fanghy,zhangc,zhugeb,zhugej.constructionofnovelrecombinantstrainscapableofproducing1,3-propanediol[j].chinjapplenvironbiol,2009,15(5):708-
|
| [8] | urrydw.physicalchemistryofbiologicalfreeenergytransductionasdemonstratedbyelasticprotein-basedpolymers[j].physchem,1997,51:11007-11028
|
| [9] | urrydw.freeenergytransductioninpolypeptidesandproteinsbasedoninversetemperaturetransitions[j].progbiophysmolbiol,1992,57:23-57
|
| [10] | limdw,kimberlytc,mackayja,chilkotia.improvednon-chromatographicpurificationofarecombinantproteinbycationicelastin-likepolypeptides[j].biomacromolecules,2007,8(5):1417-1424
|
| [11] | flossdm,schallauk,rose-johns,conradu,schellerj.elastin-likepolypeptidesrevolutionizerecombinantproteinexpressionandtheirbiomedicalapplication[j].trendsbiotechnol,2010,28:37-45
|
| [12] | trabbic-carlsonk,liul,kimb,chilkotia.expressionandpurificationofrecombinantproteinsfromescherichiacoli:comparisonofanelastin-likepolypeptidefusionwithanoligohistidinefusion[j].proteinsci,2004,13:3274-3284
|
| [13] | liw,ngis,fangbs,zhanggy.codonoptimizationof1,3-propanedioloxidoreductaseexpressioninescherichiacoliandenzymaticproperties[j].electronjbiotechnol,2011,14(4),doi:10.2225/vol14-issue4-fulltext-9
|
| [14] | ahrensk,menzelk,zenga,deckwerw.kinetic,dynamic,andpathwaystudiesofglycerolmetabolismbyklebsiellapneumoniaeinanaerobiccontinuousculture.iii.enzymesand?uxesofglyceroldissimilationand1,3-propanediolformation[j].biotechnolbioeng,1998,59:544-552
|
| [15] | 陈宏文,聂金峰,方柏山.克雷伯杆菌1,3-丙二醇氧化还原酶的分离纯化及其酶学性质[j].华侨大学报(自然科学版),2009,30:62-66[chenhw,niejf,fangbs.purificationandcharacterizationof1,3-propanedioloxidoreductasefromklebsiellapneumonia[j].jhuaqiaouniv(natsci),2009,30:62-
|
| [16] | choy,zhangy,christensent,saglelb,chilkotia,cremerps.effectsofhofmeisteranionsonthephasetransitiontemperatureofelastin-likepolypeptides[j].physchemb,2008,112(44):13765-13771
|
| [17] | gex,yangds,trabbic-carlsonk,kimb,chilkotia,filipecd.self-cleavablestimulusresponsivetagsforproteinpurificationwithoutchromatography[j].amchemsoc,2005,127:11228-11229
|
Full-Text
|
|
Contact Us
service@oalib.com QQ:3279437679 
WhatsApp +8615387084133
|
|
