耐热?α?-葡萄糖苷酶的诱导条件及重组酶的固定化
DOI: 10.3969/j.jssn.1000-2006.2012.06.018, PP. 90-94
Keywords: 耐热?α?-葡萄糖苷酶,诱导条件,酶的固定化
Abstract:
以构建重组菌(jm109/pbv-220-glz)作为耐热?α?-葡萄糖苷酶的生产菌株,研究了诱导温度、诱导时间以及重组菌生长的不同阶段对重组菌生产耐热?α?-葡萄糖苷酶的影响,并对诱导表达的重组酶进行了固定化研究。结果表明,最佳的诱导条件是:诱导温度为40℃,诱导时间为14h,在重组菌?d?(600)为2?0时诱导的酶活最高。适宜的固定化条件为:海藻酸钠20g/l,cacl?2?20g/l?,硬化时间2h,前交联剂戊二醛的质量分数为1?0%,该条件下的酶回收率高达?81%。?制备的固定化酶有较好的储存稳定性和操作稳定性,重复操作15次后,酶活为初始酶活的75%。
References
[1] | 岳振峰,陈小霞,彭志英.?α?-葡萄糖苷酶研究现状及进展[j].食品与发酵工业,2000,26(3):63-67.?
|
[2] | henrissatb.aclassificationofglycosylhydrolasesbasedonaminoacidsequencesimilarity[j].biochemj,1991,280:309-316.?
|
[3] | chibas.molecularmechanismin?α??glucosidaseandglucoamylase[j].bioscibiotechbiochem,1997,61:1233-1239.?
|
[4] | 毕金峰,魏宝东.?α?-转移葡萄糖苷酶粗酶液的酶学性质[j].食品科学,2005,26(2):75-78.?
|
[5] | costantinohr,brownsh,kellyrm.purificationandcharacterizationofanalpha?glucosidasefromahyperthermophilicarchaebacterium,?pyrococcusfuriosus?,exhibitingatemperatureoptimumof105to115degreesc[j].jbacteriol,1990,172(7):3654-3660.?
|
[6] | 郑迎迎,李大力.壳聚糖载体的制备及脲酶的固定化研究[j].生物技术,2005,15(2):56-59.?
|
[7] | 苏二正,夏涛,宛晓春,等.单宁酶和β-葡萄糖苷酶的共固定化[j].食品与生物技术学报,2006,25(1):40-44.?
|
[8] | wangyh,jiangy,duanzy,etal.expressionandcharacterizationofan?α??glucosidasefrom?thermoanaerobacterethanolicus?jw200withpotentialforindustrialapplication[j].biologia,2009,64(6):1053-1057.
|
[9] | wimmerb,lottspeichf,ritterj,etal.anoveltypeofthermostablealpha?d?glucosidasefrom?thermoanaerobacterthermohydrosulfuricus?exhibitingmaltodextrinohydrolaseactivity[j].biochemj,1997,328:581-586.?
|
[10] | 杨磊,吕明生,王淑军,等.超嗜热古菌?thermococcus?sp.hj21产高温α-葡萄糖苷酶条件和酶学性质初步研究[j].食品与发酵工业,2008,34(7):1-6.?
|
[11] | shaowl.isolationandcharacterizationofhermicellulolyticenzymesfromanaerobicthermohphiles[d].athens:universityofgeorgia,1993.?
|
[12] | marteins.proteinengineeringofcellulases[j].biochembiophyacta,2000,1543:239-252.?
|
[13] | ohtat,tokishitas,imazukar,etal.β?glucosidaseasareporterforthegeneexpressionstudiesin?thermusthermophilus?andconstitutiveexpressionofdnarepairgenes[j].mutagenesis,2006,21(4):255-260.?
|
[14] | dekkerk,yamagatah,sakaguchik,etal.xylose(glucose)isomerasegenefromthethermophile?thermusthermophilus?:cloning,sequencing,andcomparisonwithotherthermostablexyloseisomerases[j].jbacteriol,1991,17(10):3078-3083.?
|
[15] | 丁莉,许伟,严明,等.?thermusthermophilus?hb8葡萄糖异构酶在大肠杆菌中表达[j].生物加工过程,2006,4(2):42-43.?
|
[16] | quinnz,zhouk,chenxd.immobilizationof?α??galactosidaseongraphitesurfacebyglutaraldehyde[j].jfoodeng,2001,48(1):69-74.?
|
[17] | 顾旭炯,刘?,朱巍,等.海藻酸钠包埋法共固定?α?-淀粉酶和糖化酶的研究[j].化学与生物工程,2006,23(5):26-28.?
|
[18] | 杨本宏,蔡敬民,吴克,等.海藻酸钠固定化根霉脂肪酶的制备及其性质[j].催化学报,2005,26(11):977-981.?
|
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