酶法合成右旋糖酐过程规律的研究
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摘要
右旋糖酐蔗糖酶可催化蔗糖产生D-葡萄糖基,并将其转移到糖链上聚合形成不同分子量的右旋糖酐,应用于食品、医药和化工等行业。区别于菌发酵,酶法合成右旋糖酐具有高效、绿色、反应体系简单等优点。文章通过构建单酶和双酶体系,探索右旋糖酐蔗糖酶及协同右旋糖酐酶催化蔗糖制备右旋糖酐的过程规律。结果表明:在单酶体系条件下,产物右旋糖酐分子量均处于百万级别以上;同时引入右旋糖酐酶构建双酶体系,反应趋向于合成低聚右旋糖酐;随着右旋糖酐酶加入时间的延长,中等分子量片段(10~5 Da10~6 Da)右旋糖酐所占比例越大,而低分子量片段(10~4 Da Dextransucrase can catalyze the synthesis of D-glucose from sucrose and transfer it to the sugar chain to form dextran with different molecular weights.It is used in food,pharmaceutical and chemical industries.Compare to the bacterial fermentation,the enzymatic synthesis of dextran has the advantages of high efficiency,green and simple reaction system.In this paper,the process law of dextran produced by dextransucrase and dextranase is investigated by constructing single and dual enzyme systems.The results show that under the condition of single-enzyme system,the molecular weight of the product dextran is more than one million.At the same time,dextrin is introduced to construct dual-enzyme system,the reaction system tends to synthesize oligodextrans;with the increase of adding time of dextranase,the middle molecular weight fragment(10~5 Da10~6 Da)of dextran increase,while the proportion of low molecular weight fragment(10~4 Da
引文
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[21]Robyt J F.Mechanisms in the glucansucrase synthesis of polysaccharides and oligosaccharides from sucrose[J].Adv Carbohydr Chem Biochem,1995,51:133-168.
[22]Kim Y M,Seo M Y,Kang H K,et al.Construction of a fusion enzyme of dextransucrase and dextranase:application for one-step synthesis of isomalto-oligosaccharides[J].Enzyme and Microbial Technology,2009,44(3):159-164.
[2]Gan W,Zhang H,Zhang Y,et al.Biosynthesis of oligodextrans with different Mw by synergistic catalysis of dextransucrase and dextranase[J].Carbohydrate Polymers,2014,112:387-395.
[3]Goulas A K,Cooper J M,Grandison A S,et al.Synthesis of isomaltooligosaccharides and oligodextrans in a recycle membrane bioreactor by the combined use of dextransucrase and dextranase[J].Biotechnology and Bioengineering,2004,88(6):778-787.
[4]Naessens M,Cerdobbel A,Soetaert W,et al.Leuconostoc dextransucrase and dextran:production,properties and applications[J].Journal of Chemical Technology and Biotechnology,2005,80(8):845-860.
[5]Bejar W,Gabriel V,Amari M,et al.Characterization of glucansucrase and dextran from Weissella sp.TN610 with potential as safe food additives[J].International Journal of Biological Macromolecules,2013,52:125-132.
[6]李大峰,贾冬英,姚开,等.食品添加剂在复合调味品中的应用现状与展望[J].中国调味品,2011,36(1):1-4.
[7]Falconer D J,Mukerjea R,Robyt J F.Biosynthesis of dextrans with different molecular weights by selecting the concentration of Leuconostoc mesenteroides B-512FMC dextransucrase,the sucrose concentration,and the temperature[J].Carbohydrate Research,2011,346(2):280-284.
[8]Shukla S,Shi Q,Maina N H,et al.Weissella confusa Cab3dextransucrase:properties and in vitro synthesis of dextran and glucooligosaccharides[J].Carbohydrate Polymers,2014,101:554-564.
[9]邹青松.右旋糖酐分子量定向控制过程及其机理研究[D].南宁:广西大学,2013.
[10]Robyt J F,Yoon S H,Mukerjea R.Dextransucrase and the mechanism for dextran biosynthesis[J].Carbohydrate Research,2008,343(18):3039-3048.
[11]Shukla S,Verma A K,Kajala I,et al.Structure modeling and functional analysis of recombinant dextransucrase from Weissella confusa Cab3 expressed in Lactococcus lactis[J].Preparative Biochemistry and Biotechnology,2016,46(8):822-832.
[12]张磊,江帆,高亚会,等.酶催化在聚合物合成中的应用进展[J].高分子通报,2016(2):23-31.
[13]Mountzouris K C,Gilmour S G,Rastall R A.Continuous production of oligodextrans via controlled hydrolysis of dextran in an enzyme membrane reactor[J].Journal of Food Science,2002,67(5):1767-1771.
[14]Zohra R R,Aman A,Ansari A,et al.Purification,characterization and end product analysis of dextran degrading endodextranase from Bacillus licheniformis,KIBGE-IB25[J].International Journal of Biological Macromolecules,2015,78:243-248.
[15]张义平.固定化双酶法定向合成右旋糖酐及其调控机理研究[D].南宁:广西大学,2015.
[16]Hou D Z,Pu Y Y,Zou Q S,et al.Improvement of dextransucrase activity by glucose acclimation cultivation and kinetics of sucrose consumption[J].Sugar Tech,2018,20(1):60-68.
[17]刘玫.右旋糖酐蔗糖酶分离纯化方法的研究[D].南宁:广西大学,2014.
[18]Vettori M H P B,Mukerjea R,Robyt J F.Comparative study of the efficacies of nine assay methods for the dextransucrase synthesis of dextran[J].Carbohydrate Research,2011,346(9):1077-1082..
[19]Bashari M,Eibaid A,Wang J,et al.Influence of low ultrasound intensity on the degradation of dextran catalyzed by dextranase[J].Ultrasonics Sonochemistry,2013,20(1):155-161.
[20]侯殿志.酶法耦合两级膜定向制备右旋糖酐及其机理研究[D].南宁:广西大学,2017.
[21]Robyt J F.Mechanisms in the glucansucrase synthesis of polysaccharides and oligosaccharides from sucrose[J].Adv Carbohydr Chem Biochem,1995,51:133-168.
[22]Kim Y M,Seo M Y,Kang H K,et al.Construction of a fusion enzyme of dextransucrase and dextranase:application for one-step synthesis of isomalto-oligosaccharides[J].Enzyme and Microbial Technology,2009,44(3):159-164.