A1A0A2A3A4A5A6本科生畢業(yè)設(shè)計(jì)（論文）題目粗毛栓菌木聚糖酶的分離與純化姓名XXX學(xué)號(hào)XXX系別生命科學(xué)專業(yè)生物工程指導(dǎo)教師XXX職稱教授年月日教務(wù)處制A1A0A2A3A4A5A62中文摘要將粗毛栓菌木聚糖酶粗酶液經(jīng)SEPHADEXG150分子篩層析柱，最后通過(guò)活性聚丙烯酰胺凝膠電泳進(jìn)行進(jìn)一步分離純化，獲得具有木聚糖酶活性的組分。關(guān)鍵詞A7粗毛栓菌,木聚糖酶,分子篩層析，活性聚丙烯酰胺凝膠電泳ABSTRACTA8THECRUDEENZYMEFROMWHEATSTRAWCULTUREOFTRAMETESGALLICAFRWASAPPLIEDTOSEPHADEXG150GELFILTRATIONACOMPONENTWITHXYLANASEACTIVITYWASFINALLYOBTAINEDBYNATIVEPOLYACRYLAMIDEGELELECTROPHORESISKEYWORDSTRAMETESGALLICAFRA9XYLANASEA9SEPHADEXG150GELFILTRATIONA9NATIVEPAGEA10A11A12A13A14A15A163指導(dǎo)教師評(píng)語(yǔ)該生認(rèn)真查閱參考文獻(xiàn)，實(shí)驗(yàn)方案設(shè)計(jì)合理，在規(guī)定時(shí)間內(nèi)完成了預(yù)定任務(wù)，初步掌握了從事有關(guān)研究的基本實(shí)驗(yàn)技能，具備了一定的獨(dú)立從事科學(xué)研究的能力，達(dá)到了本科生的培養(yǎng)目標(biāo)。該文達(dá)到了學(xué)士學(xué)位論文水平。A17A18A19A20A21A22A23A24A25A26A27A28A29A30A29A30A29A30A31A31A32A33A34A35A36A21A37A38A23A24A25A26A39A40A41A42A43A41A42A31A32A44A37A23A24A25A26A45A46A47A48A49A50A51A52A53A54A55A53A56A57A58A59A60A61A54A62A63A60A64A65A66A67A68A66A69A70A71A72A73A74A72A75A76A67A77A78A79A80A81A82A83A84A85A86A87A88A89A81A86A89A90A91A92A93A94A95A96A86A89A97A84A85A72A98A99A100A101A72A102A103A104A105A85A72A106A107A108A109A85A72A110A111A112A111A58A113A114A90A115A106A116A67A117A118A119A94A81A120A121A93A122A123A110A1114目錄摘要1關(guān)鍵詞1ABSG87G85AG70G871G46EG92G90G82G85DS1G5353G1634011G7460G7021G994方G887311G171G7460G702111G171G171G1214G313211G171G17G21G16809G2070G211G171G17G22G9354液的G18209G2058G211G171G17G23菌G12193G211G17G21實(shí)驗(yàn)方G8873G211G17G21G171粗酶液的G11839G18252G19145G11428析G211G17G21G17G21SEPHADEXG150分子篩層析G221G17G21G17G22木聚糖酶的活性聚丙烯酰胺凝膠電泳G221G17G21G17G23木糖標(biāo)G1946G7366G13459的G8991定G231G17G21G175木聚糖酶活力的G8991定G231G17G21G17G25G15519G11345G2559G18339的G8991定G23G21G13479G7536G994分析5G21G171木糖標(biāo)G1946G7366G134595G21G17G21G15519G11345G2559G18339標(biāo)G1946G7366G13459G25G21G17G22粗毛栓菌木聚糖酶柱層析實(shí)驗(yàn)G13479G7536G25G21G17G23活性聚丙烯酰胺凝膠電泳后G7186G13479G7536G26G22G13479論G27參考文獻(xiàn)G28G14280G16886G28G1593211木糖G8999G5242G994G1821G4506G5242G1552的關(guān)G130075G32821G171木糖標(biāo)G1946G7366G13459G32825G159321G21G10287G15892G9177G15519G11345G8999G5242G994G1821G4506G5242G1552G1552的關(guān)G13007A124A124A124A1246G32821G17G21G15519G11345G2559G18339標(biāo)G1946G7366G13459G25G159321G22粗毛栓菌木聚糖酶G2520步G20600的純化G13479G7536G25G32821G17G22SEPHADEXG150柱層析G3282G16901G26G32821G17G23活性聚丙烯酰胺凝膠電泳后的G7186G14406G3282G26A120A121A93A122A123A110A1111粗毛栓菌木聚糖酶的分離和純化生G10301G5049G12255G999G1006學(xué)生G59G59G59G6363G4560G6957G5084G59G59G59摘要將粗毛栓菌木聚糖酶粗酶液經(jīng)SEPHADEXG150分子篩層析柱，最后通過(guò)活性聚丙烯酰胺凝膠電泳進(jìn)行進(jìn)一步分離純化，獲得具有木聚糖酶活性的組分。關(guān)鍵詞粗毛栓菌,木聚糖酶,分子篩層析，活性聚丙烯酰胺凝膠電泳ISOLATIONANDPURIFICATIONOFXYLANASEINTRAMETESGALLICAABSTRACTTHECRUDEENZYMEFROMWHEATSTRAWCULTUREOFTRAMETESGALLICAFRWASAPPLIEDTOSEPHADEXG150GELFILTRATIONACOMPONENTWITHXYLANASEACTIVITYWASFINALLYOBTAINEDBYNATIVEPOLYACRYLAMIDEGELELECTROPHORESISKEYWORDSTRAMETESGALLICAFRA125XYLANASEA125SEPHADEXG150GELFILTRATIONA125NATIVEPAGE引言木聚糖G7171G7905G10301G13466G14002G3733G1025G1039要的G2334G13432G13512G13044成分，G7171G14270G9994G11040G1025G1177G8437G1122G13432G13512G13044的G2499G1889生的G1028G4512生G10301G17176G9316A126A127A128A129。木聚糖G19489G16311G19668要木聚糖酶G13007的G2339G2528G1328G11004，G1039要G2265G6336G1328G11004G1122G1039G19154內(nèi)G18108木糖G14539鍵的G163G79，G23內(nèi)G2011木聚糖酶G11EG81DG82G79，G23G163G39XG92G79AG81XG92G79AG81G82HG92DG85G82G79ASE，G40G38G714G22G714G21G7141G714G27G12，將木聚糖分G16311成木G4533糖G2656G4581G18339的木糖G727G1328G11004G1122木聚糖G2656G1314聚木糖G19762G17836G2419G12483的G1631，G23G714G3818G2011木聚糖酶G708EXG82G79，G23一G163G39XG92G79AG81XG92G79AG81G82HG92DMG79ASE，G40G38G714G22G714G21G7141G714G26G12，產(chǎn)G10301為木糖G727G1328G11004G1122G1314聚木糖G19762末G12483G163木糖G14539酶G111，G23一G163G39XG92G79G82SIDASE，G40G38G714G22G714G21G7141G714G22G26G12，釋放出木糖A126A128A129。木聚糖酶在G14270G9994G11040廣泛分布，海洋及陸地G13466菌、海洋藻類、真菌、酵母、瘤胃G2656反芻動(dòng)G10301G13466菌、蝸G10287、甲殼動(dòng)G10301、陸地G7905G10301組織G2656G2520G12193無(wú)脊椎動(dòng)G10301G1025都存在。由G1122微生G10301來(lái)G9316的木聚糖G19489G16311酶普遍存在G1122G14270G9994G11040且G12193類繁多應(yīng)G11004領(lǐng)域廣泛A126A127A129。近年來(lái)，木聚糖酶在飼G7021、造紙及食品等行G1006均有廣泛的應(yīng)G11004，如紙漿生G10301漂G11345，功能性G1314聚木糖生產(chǎn)及面團(tuán)改良G2070，飼G7021添加G2070等A126A127A130A129。本文G1039要對(duì)粗毛栓菌木聚糖酶進(jìn)行了分離純化以便對(duì)其酶學(xué)性質(zhì)進(jìn)行進(jìn)一步的研究。1材料與方法A131A132A133A134A135A136A137211材料111儀器A138A139A140A141A142A143A144A145HISENSEA146A147A148A149A150A151A152A153A154A155A156A157A158A159A160A161A162A162A163A164A165A166A153A167A148A168A169A170A171A172A153A173A159A174A160A175A176A177A178A167A148A168A169A170A171A172A153A173A179A180A158A160A181A182A183A184A185A186A187A172A167A148A188A189A190A172A191A192A173A159A193A160A194A195A196A197A198A199A172A167A148A200A169A170A171A172A153A173A201A202A203A160A204A205A160A175A188A189A206A170A207A208A153A167A148A209A210A170A171A172A153A173A158A211A160A212A213A212A162A214A215A216A217A170A218A218A164A219A150A151A220A171A221A210A172A153A154A155A222A223A156A157A176A224A225A226A227A150A151A228A229A230A231A172A153A173112試劑SEPHADEXG150購(gòu)G14270PHARMACIA公司G727樺木木聚糖G708BIRCHWOODXYLAN）購(gòu)G14270美國(guó)SIGMA公司，其它G16809G2070均為國(guó)產(chǎn)分析純。113溶液的配制11313,5二硝基水楊酸（DNS）試劑甲液G1946確稱取10GNAOH，G11004去離子水G9354G16311，定容至G2100ML，將其分為50MLG2656150ML兩G18108分，在50MLNAOHG9354液G1025加入G21G苯酚，在150MLNAOHG9354液G1025加入10GG39NS，加熱G9354G16311G727乙液稱取G2200G酒石G18252鉀鈉G9354G1122500ML水G1025，加熱G9354G16311。待甲乙兩G12193G9354液冷卻后，將二者合并、混勻，加入0G175G亞G11839G18252氫鈉，攪拌、混勻，定容至1000ML。113202MOL/L醋酸鈉醋酸緩沖液PH50將G2600ML的0G17G21MG82G79/L醋G18252鈉G9354液G2656G2200ML的0G17G21MG82G79/L的醋G18252G9354液混合，G11004PH計(jì)調(diào)至PH5G170。11331樺木木聚糖溶液稱取0G175G樺木木聚糖，加入G220MLPH5G170的醋G18252鈉醋G18252緩沖液，加熱、攪拌G9354G16311，冷卻后G11004G2528樣的緩沖液定容至50ML，G23冰箱保存。1134考馬斯亮試劑稱取考馬斯亮藍(lán)GG2150100MGG9354G112250MLG285的乙G18267G1025，加入10MLG275的G11979G18252，G11004去離子水G12244釋到1000ML。114菌種本實(shí)驗(yàn)G4472保存的G10628成的具有G17751G5390G19489G16311木質(zhì)G13432G13512G13044能力的菌G7678粗毛栓菌G708TRAMETESGALLICAFR），G11345G14116G6297子菌粗毛栓菌G13007的G59G59G59G2350士G11221G28G28G26年G27G7388在G4677G1008G11477G14755G8913G5078G2283G18078的G6264G3490G3576G990，從G15999G11745G1252的G7484G7653G990分離得到的。該G18338生菌G7678G1122G2528年經(jīng)由G4677G1008G11477科學(xué)G19510生G10301研究A232A233A234A235A236A237A2383G6164微生G10301分類G4472的馬G2563G7138G1820生對(duì)其進(jìn)行了初步G18504定，定G2529為TRAMETESGALLICAA239A240A241。12試驗(yàn)方法121粗酶液的硫酸銨鹽析G2533粗酶液G1025添加G3278G1319G11NHA240G12A242SOA240至G220G20293G2656G5242，G23過(guò)G3824，離G5527G708G25,000G85/MIG81，G23）15MIG81，G19512去G7446G15519G11345G8797G9108G10301G727G9994后將G990G9177液的G11839G18252G19145G20293G2656G5242G990調(diào)至G265，G1889G1122G23G19757G13634過(guò)G3824，離G5527G708G26,500G85/MIG81，G23）10MIG81，G5335G990G9177，得G8797G9108G13434100ML，將G8797G9108G1122G210冰箱G1025保存，備G11004。將G8797G9108G17891析G708G6142G11053G183391G211G23G46G39），G8611G19560G22G23G4579時(shí)G6454一G8437G23去離子水，G11464到G17891析G15967內(nèi)酶液無(wú)G8543G11053的G11NHA240G12A242SOA240。G17891析液經(jīng)聚乙二G18267G708平均分子G18339G730G210,000）G8999G13565后，G1328為G991一步進(jìn)行分子篩層析的樣品。122SEPHADEXG150分子篩層析16CM115CM將G3800理G3921的凝膠放在G3835G9915G7491G1025，G11004去離子水反G3809沖G8939至G1025性，G1889G1100450MMG82G79/L的醋G18252鈉緩沖液G708PH5G170）沖G8939G1972G8437。將凝膠G16025G2058成1G17G25G104115G70M柱，G990樣G2081G11004G990G17860緩沖液平G15925柱，過(guò)G3824。G990樣時(shí)樣品G1319G12227G993G4464過(guò)多，G13434為柱G1319G12227的15。G8939G14085G13459G17907G5242為G25G70M/H，G8611G12661G6922G19610G21ML。G7828G8991G8981出液的酶活性，合并具有G11468G2528酶活性的組分，G9994后G1889如G990G8873進(jìn)行G11NHA240G12A242SOA240G11428析、離G5527G2656G17891析等步G20600A239A243A241。123木聚糖酶的活性聚丙烯酰胺凝膠電泳活性聚丙烯酰胺凝膠電泳G708G81AG87IG89EG51AGG40）A239A244A245A246A246A241G11004G1122木聚糖酶的G3250G6922G2058備及活性G7828G8991，樣品為經(jīng)過(guò)層析G8873分離的G18108分純化的酶樣品。G27分離膠的G2058備G708G210ML）取5G17G22MLG220凝膠G1660備液G708G2559G21G28丙烯酰胺G26561N,NG255亞甲G2464丙烯酰胺）、G21G175MLG22G170MG82G79/LG55G85ISHG38G79緩沖液G708PHG27G17G28）、0G171ML10G55G40G48G40G39、0G17G21ML10過(guò)G11839G18252G19145、11G17G28MLDHA242O，混勻，G9760入G39G60G60G2675G3423G3414G11464G7507電泳G8145的G10639G10839G7507間G19565G1025，G1363液膠的G20652G5242為G10639G10839G7507G13449G19283的G21/G22G22/G23。G990面G4565一G15192水層。G1363G10639G10839G7507在G220G5670G9213箱G1025聚合G210G220MIG81。5G8999G13565膠的G2058備G1110MLG12取1G17G26MLG220凝膠G1660備液、G21G175ML0G175MG82G79/LG55G85ISHG38G79緩沖液G708PHG25G17G26）、0G1705ML10G55G40G48G40G39、0G171ML10過(guò)G11839G18252G19145、5G17G255MLDHA247O，混勻，加G1122分離膠G990面的G12366G19565G1025。在G8999G13565膠的G990面G11053有1G21G70MG20652的G12366G19565G708G990樣G11004），G1889如G990G8873進(jìn)行G5670G9213聚合。進(jìn)行G2058備電泳時(shí)，將酶液G994G25G104加樣緩沖液G708G2559PHG25G17G26的0G17G265MG82G79/LG55G85ISHG38G79、G220G10988G8845、0G1701G9352酚藍(lán)）混合，G990樣后，G2045G11004PHG27G17G22的G55G85ISG10988G8700G18252緩沖G13007G13491G708G215MMG82L/LG55G85IS、G2150MMG82G79/LG10988G8700G18252），G1122G23進(jìn)行G5670G8981G70810MA）電泳，待樣品進(jìn)入分離膠后，G1889將電G8981G2331至15MA，G13499G13505電泳。電泳至G9352酚藍(lán)G6363G12046G2070離膠G5225面1G21G70M時(shí)G1584止電泳。取出并分離G10639G10839G7507，并G1363凝膠附著在一塊G10639G10839G7507G990。G11004刀片G13449G2533G2011去凝膠兩邊的G18108分G708G8611邊G134341G70M寬）。由G1122酶樣品在凝膠的兩邊沿G3800往往略呈弓形，G993G4464G11004來(lái)進(jìn)行酶的定位，故將凝膠的兩邊G18108分G2011G19512。G1889G11004刀片沿凝膠一側(cè)G13449G2533G2011去寬G134341G70M的膠條，將此膠A248A249A250A251A252A253A2544條的分離膠G18108分從G990到G991G2011成G21MMG19283的膠塊，放在G11468應(yīng)編號(hào)的G40G51G12661G1025，搗碎膠塊，G9994后G2533G8611個(gè)G40G51G12661G1025分別加入50L的1樺木木聚糖，G13634G1122G235水浴鍋G1025保G92131G175H，G1889G2533G8611個(gè)G40G51G12661G1025分別滴加100L1G39NS，沸水浴煮15MIG81。觀察G2520G12661G1025顏G14406變化。進(jìn)行G7186G14406反應(yīng)期間，G11004保鮮膜覆蓋剩余膠塊，并G13634G1122G23冰箱G1025暫時(shí)保存。124木糖標(biāo)準(zhǔn)曲線的測(cè)定G1946確稱取干燥至G5670重的木糖標(biāo)G1946品50MG，G11004蒸餾水G9354G16311，定容至50ML。按G991G15932在G2520G12661G1025加入G2520G12193G16809G2070A255A109A111A0A113A1A115A2A90A53A116A83A41A117A3A4A109A17A120A5A19A42A42A6A53A42A6A83A42A6A7A42A6A8A90A6A42A124A3A9A10A17A120A52A19A42A42A6A53A42A6A83A42A6A7A42A6A8A90A6A42A127A11A12A17A120A52A19A53A6A42A90A6A8A90A6A7A90A6A83A90A6A53A90A6A42A71A13A14A130A15A17A120A52A19A90A6A41A90A6A41A90A6A41A90A6A41A90A6A41A90A6A41將G990G17860G16809G2070加完后，混勻G2520G12661，沸水浴5MIG81，冷卻至G4472G9213，分別G11004去離子水將G2520G12661定容至G215ML，混勻。選G11004G1821G122551G70M的比G14406G7491在550G81MG3800比G14406，G11004G12366G11345G12661G1328對(duì)照，G8991G2520G12661G1821G4506G5242G1552，G8611G12661重G3809三G8437，求平均G1552。以木糖G8999G5242為橫坐標(biāo)，G1821G4506G5242G1552為G13449坐標(biāo)，繪G2058標(biāo)G1946G7366G13459A16A18A20。125木聚糖酶活力的測(cè)定采G11004G39NSG708DIG81IG87G85G82SAG79IG70G92G79IG70AG70ID）G8873即G22,5二硝基水G7484G18252G8873A16A21A22A23A24A20。取0G17G28ML由50MMG82G79/LNAAG70HAG70G708PHG23G175）緩沖液G18209G2058的1樺木木聚糖G1122G215ML刻G5242G16809G12661G1025，加入0G171ML適當(dāng)G12244釋的酶液，G1122G235水浴G1025保G9213G220MIG81后，加入G22ML1G39NSG16809G2070，混勻，在沸水浴G1025煮15MIG81，冷卻至G4472G9213，補(bǔ)加水至G215ML，混勻，G1122550G81MG3800G8991定吸G1821G1552。以G8611分鐘催化木聚糖水G16311生成1MG82G79木糖G6164G19668的酶G18339定為1個(gè)活力單位。木聚糖酶酶活力計(jì)算公式如G991木聚糖酶酶活力G708UMIG81A25A23MLA25A23）G81A/G708150G171G22G104G220）其G1025，G81為酶液G12244釋倍數(shù)，A為標(biāo)G1946G7366G13459G990吸G1821G1552G6164對(duì)應(yīng)的木糖微克數(shù)，150G171G22為木糖分子G18339，G220為酶G1328G11004時(shí)間G708MIG81）。126蛋白含量的測(cè)定參照BG85ADFG82G85DA16A26A20G8873取一G13007列G16809G12661，按G991G15932分別加入G991列G16809G2070A248A249A250A251A252A253A2545A115A2A90A53A116A83A41A79A14A132A17A90A42A42A27A5A28A120A52A19A42A42A6A53A42A6A83A42A6A7A42A6A8A90A6A42A135A29A30A31A32A140A40A53A41A42A17A120A52A19A41A41A41A41A41A41A127A11A12A17A120A52A19A90A6A42A42A6A8A42A6A7A42A6A83A42A6A53A42A6A42將G990G17860G16809G2070加完后，混勻，G4472G9213G19757G13634G21MIG81，選G11004G1821G12255為1G70M的比G14406G7491，以G993加G10287G15892G9177G11345G15519G11345G11BSAG12的G16809G12661為對(duì)照，在5G285G81MG3800比G14406。以G10287G15892G9177G11345G15519G11345的G8999G5242為橫坐標(biāo)，G1821G4506G5242G1552為G13449坐標(biāo)，繪G2058標(biāo)G1946G7366G13459。取待G8991樣品G9354液1ML，加入考馬斯亮藍(lán)GG21505ML，混勻，按G990G17860方G8873G8991定5G285G81M的G1821G4506G5242G1552。平行做G22份。比G14406后，從標(biāo)G1946G7366G13459G990查出待G8991樣品的G15519G11345質(zhì)G8999G5242。2結(jié)果與分析21木糖標(biāo)準(zhǔn)曲線在G990G17860條件G991，得木糖標(biāo)G1946品G8999G5242G994G1821G4506G5242G1552的關(guān)G13007G708G1593211）。A3311A34A35A36A37A38A39A43A37A44A45A46A47A124A3A48A49A50MG/ML020406081A96A51A54A55A17OD550A1901010280046406220747以木糖G2559G18339為橫坐標(biāo)，G1821G4506G5242G1552為G13449坐標(biāo)，繪G2058出標(biāo)G1946G7366G13459G708G32821G171）。計(jì)算出標(biāo)G1946G7366G13459方G12255為A0G17G2715X0G170G23G26G708RA2470G17G28G28G23）G13479G7536G15932G7138，在00G17G27MG/ML范圍內(nèi)木糖G8999G5242G994G1821G4506G5242G1552G13459性關(guān)G13007良G3921。A56A57A58A59A60A61A62A63A64A65A6622蛋白含量標(biāo)準(zhǔn)曲線在G990G17860條件G991，得G10287G15892G9177G15519G11345G8999G5242G994G1821G4506G5242G1552的關(guān)G13007G708G159321G21）A67A68A69A70A72A73A74A75A76A77A74A78A78A80A81A82BSAA146A84A85G/MLA8620406080100A87A88A84A89A85OD595A8602360469067607880906A91A92A93A94A95A97A98A99A100A93A94A93A101A102A102A103A104A92A93A94A105A105A101A95A93A93A94A98A97A93A93A94A106A93A94A101A93A94A107A93A94A95A97A97A94A106A108A110A112A114A114A118A118MG/MLA119A121A122A123A125A126OD550A128A129A131A133A134A136A137A138A139A141A142A143A144A1456以G10287G15892G9177G11345G15519G11345G8999G5242為橫坐標(biāo)，G1821G4506G5242G1552為G13449坐標(biāo)繪G2058出標(biāo)G1946G7366G13459G708G32821G17G21）。得到標(biāo)G1946G7366G13459的G3250歸方G12255為A0G1700G27G215X0G1711G26G11RA1470G17G28G261G12，G13479G7536G15932G7138，G10287G15892G9177G11345G15519G11345G18339在0G17G211MG/ML范圍內(nèi)G994G1821G4506G5242G1552G13459性關(guān)G13007良G3921G708G32821G17G21）。A148A149A150A151A152A153A154A155A156A157A158A15923粗毛栓菌木聚糖酶柱層析實(shí)驗(yàn)結(jié)果木聚糖酶粗酶液經(jīng)過(guò)G11428析離G5527，分子篩層析等純化步G20600G13479G7536見(jiàn)G1593213。A16013A161A162A163A164A165A166A167A168A169A170A171A172A173A174A175A176TABLE13SUMMARYOFTHEPURIFICATIONOFXYLANASEINTRAMETESGALLICAA177A178A179A180A181A182A183/UA181A184A185/MGA186A182A183A187U/MGA188A189A190A191A177A178A192A193PURIFICATIONTOTALTOTALSPECIFICRECOVERYPURIFICATIONSTEPACTIVITYPROTEINACTIVITYRATEFOLDA194A195A196A197A198454A199105100A1991044541001NH42SO4FRACTIONATIONA200A201A202A203A204286A199104225A1991021271163281SEPHADEXG150MOLECULARSIEVINGA194A195A196A197A198248A1991033596689660051519NH42SO4FRACTIONATIONSEPHADEXG150分子篩層析G8939G14085G7366G13459見(jiàn)G328213。A205A206A207A208A207A207A209A210A211A212A213A207A208A214A214A215A216A217A218A206A207A208A219A215A214A211A207A207A208A211A214A210A207A220A207A221A207A209A207A214A207A207A222A223A224A225A226A225A227A228A229UG/MLA230A231A232A233A234A235OD595A236A237A238A239A240A241A242A2437A244A245A246A247SEPHADEXG150A248A249A250A244A251FIG13ELUTIONCURVEOFTHESEPHADEXG150CHROMATOGRAPHY24活性聚丙烯酰胺凝膠電泳后顯色結(jié)果按照1G171G17G23G1025G6164G17860方G8873得到的G13479G7536如G32821G17G23G6164G12046A252A253A254A255A45A0A1A2A3A4A5A6A7A8A9A10A11A12A13A252從G7186G14406G13479G7536G990G2499以看出，1號(hào)到G25號(hào)的G40G51G12661的顏G14406比其它G2520G12661的顏G14406深，即分離膠G990從負(fù)極到正極第G211G21MM的位G13634G990有酶存在G727而第G210到第G21G27G12661的顏G14406也比其他它的G2520G12661的顏G14406深，G2499能G7171G9352酚藍(lán)G6363G12046G2070的干擾。3討論本G16809驗(yàn)從粗毛栓菌的木聚糖酶粗酶液G1025,經(jīng)過(guò)G11428析，離G5527G2656分子篩層析等操G1328后,G13479G7536得到一G12193木聚糖酶的混合組分，木聚糖酶粗酶液得到初步分離G2656純化。首G1820經(jīng)過(guò)G11839G18252G19145G11428析得到的組分經(jīng)SEPHADEXG150分子篩層析，此階段酶的純化倍數(shù)G7171G21G17G271，G3250G6922率G7171G25G17G22G727G1889經(jīng)過(guò)G11428析后，酶的純化倍數(shù)G2656G3250G6922率分別G717115G171G28G26560G1705。得到的組分通過(guò)活性聚丙烯酰胺凝膠電泳G2499以G11004G1122進(jìn)一步的分離純化G2656G3250G6922。經(jīng)SEPHADEXG150分子篩層析后得到的酶液純化倍數(shù)得到很G3835的提G20652，但G3250G6922率很G1314，酶的損失很G3835，并且純G5242G2656比活力均G1314G1122范美霞A14A15A16A17的G11468應(yīng)G13479G7536，其G2419因G2499能G7171G990樣G18339G17751G3835，造成樣品分布過(guò)廣且未充分G3250G6922。另G3818，分離純化步G20600過(guò)G1122單一，G1363該實(shí)驗(yàn)未能達(dá)到理想的純化效G7536。0100200300400500600A18A19A20A21A22A21A23A21A21A23A24A25A23A25A21A23A26A21A23A20A23A23A22A19A24A21A21A24A24A21A24A19A21A27A28A29A30A31MLA32A33A34A35UA36000501015020250303504A37A38A39A40A41A42A43A44A46A47A48A49A50A51A52A538參考文獻(xiàn)A54A55A56A57A58A59A60A61A62A62A63A60A64A65A66A67A68A69A70A71A72A73A74A75A76A77A78A79A80A60A81A76A82A83A84A85A86A87A88A89A61A90A91A55A92A93A94A55A92A88A60A54A61A56A95A96A60A61A62A62A63A60A97A98A99A100A101A68A69A70A94A67A68A69A70A102A100A101A68A69A70A94A103A104A70A105A106A70A78A107A108A109A110A71A111A70A84A112A113A114A115A116A117A118A84A119A120A121A122A86A93A94A93A60A54A87A56A123A124A60A61A62A62A125A60A126A127A128A97A98A99A67A68A69A70A78A129A108A102A70A84A130A131A132A133A60A134A135A81A76A118A84A136A120A121A122A86A61A94A61A60A54A137A56A138A138A138A115A139A140A141A115A142A143A144A60A61A62A62A61A60A145A146A147A148A149A150A151A152A67A131A153A154A155A78A156A157A132A133A86A64A65A66A84A158A159A86A61A61A89A55A90A91A61A137A94A61A125A60A54A93A56A160A161A162A115A163A164A165A115A166A167A168A169A170A171A172A173A174A175A176A177A178A179A180A181A182A183A184A185A186A186A187A188A189A190A