生物转化合成右旋糖酐
摘要
右旋糖酐是一种由若干葡萄糖脱水形成的无毒无害的高分子聚合物,是世界上第一个工业化生产的微生物多糖,是最早使用的一种优良的血浆代用品,在医药、食品和石油等方面都有着广泛的应用。
本课题选用中国工业微生物菌种保藏中心的肠膜明串珠菌20074,以蔗糖为底物发酵制备右旋糖酐。研究了发酵液中右旋糖酐沉析条件及右旋糖酐分子量的测定方法,菌种的培养条件,发酵工艺条件,并初步探索了固定化菌体的方法制备右旋糖酐。
获得的最适合肠膜明串珠菌20074生长的培养基组成为:蔗糖10%、蛋白胨0.2%、磷酸氢二钠0.2%、磷酸二氢钾0.03%、酵母浸膏0.5%。获得的最佳发酵条件为:种子液发酵时间10-12h,接种量约10%,培养基pH值7.0-7.5,培养温度28℃,转速为50r/min,发酵时间20-24h,底物浓度15%,维生素E和Mn~(2+)有利于右旋糖酐的形成,配置成一定浓度溶液作为添加剂加入到发酵培养基中,加入量为0.8mL/L。用高效凝胶渗透色谱法测定发酵液中右旋糖酐分子量大于40万,用红外光谱表征产品结构中的特征基团与标准品相符。
活性炭是较好的固定化菌体的载体,肠膜明串珠菌20074在活性炭上的最佳固定化条件为:培养基初始pH值为7.10,接种量10%,固定化时间10-13h,摇床转速150r/min,固定化温度25℃。固定化菌体方法制备所得的右旋糖酐与直接发酵法制得的产品在分子量和结构上都相同,但产率有所降低。
Dextran is a kind of polymerization composing by glucose dehydrating,is of innocuity,is the first microbiology polysaccharid produced in industry, is the substitute of blood plama used at first,is used widely in medicament, food and petroleum.
In this subject, Leuconostoc mesenteroides 20074 bought from CICC is choosed,preparing dextran by fermenting when sucrose is used as substrate.The culture condition,precipitating condition of dextran in fermenting culture medium, method of measuring molecular weight of dextran and fermenting condition are investigated, research of preparing dextran by immobilizing Leuconostoc mesenteroides 20074 is done primarily.
The optimal composition of Leuconostoc mesenteroides 20074 culture medium is: sucrose 10%, peptone 0.2%, Na_2HPO_4 0.2%, KH_2PO_4 0.03%, yeast extract 0.5%.The best technology condition: fermenting time of Leuconstoc Mesenteroides 20074 10-12h, transferring content 10%, pH of culture medium 7.0-7.5, culturing temperature 28℃, vibrator speed 50r/min, fermenting time 24h, consistency of substrate 15%, vitamine E and Mn~(2+) is available for dextran forming and content of vitamine E and Mn~(2+) with certain consistency is 8mL/L. The molecular weight of dextran producd by fermenting is more than 400,000 measured by HPGPC, and the IR spectrum of dextran is accordant with the IR spectrum of standard sample.
Active carbon is a better carrier,the best technology condition of immobilizing Leuconostoc mesenteroides 20074 on active carbon is: pH of culture 7.1, transferring content 10%,immobilizing time 10-13h, vibrator speed 150r/min,immobilizing temperature 25℃.The molecular weight and structure of dextran is accordant with dextran preparing by fermenting, and the yield rate of dextran decreases.
本课题选用中国工业微生物菌种保藏中心的肠膜明串珠菌20074,以蔗糖为底物发酵制备右旋糖酐。研究了发酵液中右旋糖酐沉析条件及右旋糖酐分子量的测定方法,菌种的培养条件,发酵工艺条件,并初步探索了固定化菌体的方法制备右旋糖酐。
获得的最适合肠膜明串珠菌20074生长的培养基组成为:蔗糖10%、蛋白胨0.2%、磷酸氢二钠0.2%、磷酸二氢钾0.03%、酵母浸膏0.5%。获得的最佳发酵条件为:种子液发酵时间10-12h,接种量约10%,培养基pH值7.0-7.5,培养温度28℃,转速为50r/min,发酵时间20-24h,底物浓度15%,维生素E和Mn~(2+)有利于右旋糖酐的形成,配置成一定浓度溶液作为添加剂加入到发酵培养基中,加入量为0.8mL/L。用高效凝胶渗透色谱法测定发酵液中右旋糖酐分子量大于40万,用红外光谱表征产品结构中的特征基团与标准品相符。
活性炭是较好的固定化菌体的载体,肠膜明串珠菌20074在活性炭上的最佳固定化条件为:培养基初始pH值为7.10,接种量10%,固定化时间10-13h,摇床转速150r/min,固定化温度25℃。固定化菌体方法制备所得的右旋糖酐与直接发酵法制得的产品在分子量和结构上都相同,但产率有所降低。
Dextran is a kind of polymerization composing by glucose dehydrating,is of innocuity,is the first microbiology polysaccharid produced in industry, is the substitute of blood plama used at first,is used widely in medicament, food and petroleum.
In this subject, Leuconostoc mesenteroides 20074 bought from CICC is choosed,preparing dextran by fermenting when sucrose is used as substrate.The culture condition,precipitating condition of dextran in fermenting culture medium, method of measuring molecular weight of dextran and fermenting condition are investigated, research of preparing dextran by immobilizing Leuconostoc mesenteroides 20074 is done primarily.
The optimal composition of Leuconostoc mesenteroides 20074 culture medium is: sucrose 10%, peptone 0.2%, Na_2HPO_4 0.2%, KH_2PO_4 0.03%, yeast extract 0.5%.The best technology condition: fermenting time of Leuconstoc Mesenteroides 20074 10-12h, transferring content 10%, pH of culture medium 7.0-7.5, culturing temperature 28℃, vibrator speed 50r/min, fermenting time 24h, consistency of substrate 15%, vitamine E and Mn~(2+) is available for dextran forming and content of vitamine E and Mn~(2+) with certain consistency is 8mL/L. The molecular weight of dextran producd by fermenting is more than 400,000 measured by HPGPC, and the IR spectrum of dextran is accordant with the IR spectrum of standard sample.
Active carbon is a better carrier,the best technology condition of immobilizing Leuconostoc mesenteroides 20074 on active carbon is: pH of culture 7.1, transferring content 10%,immobilizing time 10-13h, vibrator speed 150r/min,immobilizing temperature 25℃.The molecular weight and structure of dextran is accordant with dextran preparing by fermenting, and the yield rate of dextran decreases.
引文
[1]李艳,陈学武,牟德华.右旋糖酐的生产及应用[J].山西食品工业,1998,(3):36,37,42.
[2]张洪斌.海藻酸钠固定化酶制备右旋糖酐的研究[D].安徽:合肥工业大学,2004。
[3]章朝辉.右旋糖酐的制备及应用[J].四川化工与腐蚀控制,2001,4(1):50-52.
[4]Kelly W J,Asmundson R V,Harrison G L.Differentiation of dextran-producing Leuconostoc strains from ferment rice cake(puto)using pulsed-field gel electrophoresis[J].Food Microbiology,1995,26:345-352.
[5]陈代杰,金飞燕.生物高分子(第五卷 多糖Ⅰ-原核生物多糖)[M].北京:化学工业出版社,2004:305-320.
[6]刘卫东,张启祥.人工胶体血浆代用品的分类和临床应用比较[J].中国药业,2006,15(12):62-63.
[7]章朝辉.蔗糖化学衍生物[J].精细石油化工进展,2000,1(9):8-13.
[8]章思规.精细有机化学品技术手册(上册)[M].北京:科学出版社,1992:583-584.
[9]原正平,王汝龙.化工产品手册[M].北京:化学工业出版社,1987:547-548.
[10]国家药典委员会.中华人民共和国药典(二部)[Z].北京:化学工业出版社,2005:88-90.
[11]廖威.利用生物技术防治甘蔗糖厂“蔗饭”问题的基础研究[D].广西:广西大学:2002.
[12]张力田.碳水化合物化学[M].北京:轻工业出版社,1988:377.
[13]王少武,姚日生.右旋糖酐铁的合成新工艺[J].安徽化工,2000,(5):12-14.
[14]高永锐,杨祥开,管忠谊,等.右旋糖酐铁合成工艺改进实验[J].广西畜牧兽医,2003,19(4):151.
[15]宋春和.右旋糖酐铁制备方法探讨[J].实用医技杂志,1994,4(7):526-527.
[16]范慧红,刘金秀,徐康森.高效凝胶渗透色谱在右旋糖酐铁质控中的应用[J].药物分析杂志,2000,20(4):254-257.
[17]王少武,姚日生.右旋糖酐铁合成新工艺[J].安徽化工,2003,(5):12-13.
[18]Seo E S,Lee J H,Park J Y,et al.Enzymatic synthesis and anti-coagulant effect of salicin analogs by using the Leuconostoc mesenteroides glucansucrase acceptor reaction[J].Journal of Biotechnology,2005,117:31-38.
[19]江幼岷,陈仕儒,马恩普,等.造血干细胞动员剂硫酸葡聚糖的毒性及其含硫量的关系[J].实验血液学杂志,1995,3(1):74-77.
[20]Kim D,Robyt J F.Dextransucrase constitutive mutant of Leuconostoc mesenteroides B-1299[J].Enzyme microbial Technology,1995,17:1050-1056.
[21]葛崇涛,田星,高莉莉.肠膜明串株菌生长特性研究[J].中国乳品工业,2005,(7):72-73.
[22]Kim D,Robyt J F.Production and selection of mutant of Leuconostoc mesenteroides constitutive for glucansucrase[J].Enzyme Microbial Technology,1994,16:659-664.
[23]Kim K,Kim D W,Lee J H,et al.Development of constitutive dextransucrase hyper-producing mutants of Leuconstoc mesenteroides using the synchrotron radiation in the 70-1000 eV region[J].Biotechnology Technology,11:319-321.
[24]Kitaoka M,Robyt J F.Use of a microtiter plate screening method for obtaining Leuconostoc mesenteroides mutant constitutive for glucansucrase[J].Enzyme Microbial Technology,1998,22:527-531.
[25]Santos M,Teixeira J,Rodrigues A.Production of dextransucrase,dextran and fructose from sucrose using Leuconostoc Mesenteroides NRRL B512F[J].Biochemical Engineering Journal,2000,4:177-188.
[26]Yoon S H,Fulton D B,Robyt J F.Enzymatic synthesis of two salicin analogues by reaction of salicyl alcohol with Bacillus macerans cyclomaltodextrin glucanyltransferase and Leuconostoc mesenteroides B-742CB dextransucrase[J].Carbohydrate Research,2004,339:1517-1529.
[27]宋明,金中,徐兰,等.乳酸细菌葡聚糖蔗糖酶研究应用与进展[J].检验医学教育,2005,12(2):30-32.
[28]Zivomir P,Suzana R,Dragan P,et al.Immobilization of dextran- sucrase on regenerated benzoyl cellulose carriers[J].Enzyme and Microbial Technology,2007,40:763-768.
[29]Bertrand A,Morel S,Lefoulon F,et al.Leuconostoc mesenteroides glucansucrase synthesis of flavonoid glucosides by acceptor reactions in aqueous-organic solvents[J]. Carbohydrate Research,2006,341:855-863.
[30]Ara'nzazu G S,Miguel A,Manuel B,et al.Synthesis of methyl-d-glucooligos accharidesbyen trapped dextransucrase from Leuconostoc mesenteroides B-1299[J].Journal of Biotechnology,2006,124:439-445.
[31]Monchois V,Willemot R M,Monsan P,et al.Glucansucrases:mechanism of action and structure-function relationships[J].FEMS Microbiology Reviews,1999,23(2):131-151.
[32]邵彦春,王建华,滕达,等.右旋糖酐蔗糖酶基因的克隆及其序列分析[J].微生物学报,2005,32(3):213-23.
[33]Robyt J F,Kimble B K,Walseth T F.The mechanism of dextransucrase action Direction of biosynthesis[J].Biochem.Biophys,1974,165(2):634-640.
[34]廖威.葡聚糖产生菌发酵培养基优化初步研究[J].广西科学院学报,2003,19(3):125-128.
[35]王勇,张文革,刘石.右旋糖酐制取新工艺研究[J].生物技术,1994,4(3):45-46.
[36]崔益清.右旋糖酐生产新工艺-发酵液水解法[J].药物研究,1999,8(6):17.
[37]张洪斌,姚日生,朱慧霞,等.发酵法生产右旋糖酐的工艺研究[J].合肥工业大学学报,2004,783-787.
[38]李清娣,高歌.右旋糖酐工艺与影响质量的因素[J].中国医学工业杂志,1994,25(2):91-92.
[39]井明冉.右旋糖酐生产过程蛋白胨用量探讨[J].石河子科技,2000,(5):28-29.
[40]廖威,杨辉,梁海秋,等.葡聚糖产生菌的分离鉴定与发酵条件的初步研究[J].食品科学,2003,24(11):56-59.
[41]宋学宁,庄丽萍,傅茂东.影响右旋糖酐质量的因素分析[J].山东医药工业,2003,22(6):46-47.
[42]王冶业,刘晓凤,魏甲乾.固定化肠膜明串珠菌生产右旋糖酐的研究[J].甘肃工业大学学报,2003,29(3):79-81.
[43]郭德祥.关于提高右旋糖酐发酵产率的探讨[J].山东医药工业,1994,13(2):22-23.
[44]郭杰炎,蔡武城.微生物酶[M].北京:科学出版社,1986:172-224.
[45]Sinha J,Dey P K,Panda T.Extractive fermentation for improved production of endoglucanase by an intergeneric fusant of Trichoderma reesei/Saccharomyces cerevisiae using aqueous two-pHase system[J].Biochemical Engineering Journal,2000,6:163-175.
[46]Mase T,Matsumiya Y,mori S,et al.Purification and Characterization of a Novel Glucoamylase from Acremonium sp.YT-78[J].Journal of Fermentation and Bioengineering,1996,81(4):347-350.
[47]Ivanova V,Yankov D,Kabaivanova L,et al.Simultaneous biosynthesiS and purification of two extracellular Bacillus hydrolases in aqueous two-phase systems[J].MiCrObiology Research,2001,156:19-30.
[48]Kitaoka M,Robyt J F.Use of a microtiter plate screening method for obtaining Leuconostoc mesenteroides mutants constitutive for glucansucrase[J].Enzyme and Microbial Technology,1998,22:527-531.
[49]Santos M,Rodrigues A,TeiXeira J A.Production of dextran and fructose from carob pod extract and cheese whey by Leuconostoc mesenteroides NRRL B512F[J].Biochemical Engineering Journal,2005,25:1-6.
[50]郭振友,许丹枫,吴明.超滤技术代替乙醇沉淀粗右旋糖酐工艺研究[J].中国纺织大学学报,2000,26(2):100-102.
[51]Ko E A,Nam S H,Jin X J,et al.Synthesis of thermo- and acid-stable novel oligosaccharides by using dextransucrase with high concentration of sucrose[J].Journal of Applied Glycoscience,2007,54(2):147-155.
[52]Christian T J,Merilyn M H,Richards G N.A preliminary,study of the use of larch arabinogalactan in aqueous two-pHase systems[J].Carbohydrate Polymers,1998,35:7-12.
[53]Kelly W J,Asmundson R V,Harrison G L,et al.Differentiation of dextran-producing Leuconostoc strains from fermented rice cake(puto)using pulsed-field gel electrophoresis[J].International Journal of Food Microbiogy,1995,26:345-352.
[54]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and Glucosyltransferase activity[J].Carbohydrate Research,2004,339:1029-1034.
[55]Rodrigues S,Lona M F,Franco T T.Optimizing panose production by modeling and simulation using factorial design and surface response analysis[J].Journal of Food Engineering,2006,75:433-440.
[56]Cote' G L,Sheng S.Penta-,hexa-,and heptasaccharide acceptor products of alternansucrase[J].Carbohydrate Research,2006,341(2):2066-2072.
[57]Kawakita H,Seto H,Ohto K,et al.Pore control with dextran generated from immobilized dextransucrase[J].Biochemical Engineering Journal,2007,36:190-193.
[58]Eom H J,Seo D M,Han N S.Selection of psychrotropHic Leuconostoc spp.producing highly active dextransucrase from lactate fermented vegetables[J].International Journal of Food Microbiology,2007,117:61-67.
[59]Goulas A K,Fisher D A.,Grimble G K.,et al.Synthesis of isomaltooligosaccharides and oligodextrans by the combined use of dextransucrase and dextranase[J].Enzyme and Microbial Technology,2004,35:327-338.
[60]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and glucosyltransferase activity[J].Carbohydrate Research,2004,339:1029-1034.
[61]Richard G,Yu S,Monsan P.A novel family of glucosyl 1,5-anhydro-D- fructose derivatives synthesised by transglucosylation with dextransucrase from Leuconostoc mesenteroides NRRL B-512F[J].Carbohydrate Research,2005,340:395- 401.
[62]Funane K,Ishii A,Ono H,et al.Changes in linkage pattern of glucan products induced by substitution of Lys residues in the dextransucrase[J].FEBS Letters,2005,579:4739-4745.
[63]Kang H K,Seo M Y,Seo E S,et al.Cloning and expression of levansucrase from Leuconostoc mesenteroides B-512 FMC in Escherichia coli[J].Biochimica et BiopHysica Acta,2005,1727:5-15.
[64]杨敏.酶法生产右旋糖酐[P],C12P 19/08.2000,11,02.
[65]姚日生,董明辉,朱慧霞.酶法合成右旋糖酐相对分子质量的控制[J].生物工程,2007,24(7):964-967.
[66]刘汝宽,李昌珠,张玉军.树脂固定脂肪酶技术的研究[J].河南工业大学学报(自然科学版),2007,28(3):68-71.
[67]Berensmeier S,Ergezinger M,Bohnet M,et al.Design of immobilised dextransucrase for fluidised bed application[J].Journal of Biotechnology,2004,114(3):255-267.
[68]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and glucosyltransferase activity[J].Carbohydrate Research,2004, 339:1029-1034.
[69]Kubik C,Sikora B,Bielecki S.Immobilization of dextransucrase and its use with soluble dextranase for glucooligosaccharides synthesis[J].Enzyme and Microbial Technology,2004,34:555-560.
[70]Kitaoka M,Robyt J F.Mechanism of the action of Leuconostoc Mesenteroides B-512FMC dextransucrase:kinetics of the transfer of D-glucose to maltose and the effect of enzyme and substrate concentrations[J].Carbohydrate Research,1999,320:183-191.
[71]陈建龙,祁建城,曹仪植.固定化酶研究进展.化学与生物工程[J],2006,23(2):7-9.
[72]袁定重,张秋禹,侯振宇,等.固定化酶载体材料的最新研究进展[J].材料导报,2006,20(1):69-72.
[73]范友华,黄军,陈泽君.固定化酶载体的研究现状及其应用[J].湖南林业科技,2007,34(2):29-31.
[74]Zafar I,Barker P E.An experimental and computational study of a biochemical polymerisation reaction in a chromatographic reactor separator[J].Chemical Engineering Science,1988,43(9):2369-2375.
[75]Barker P E,Ganetsos G.,Ajongwen J,et al.Bioreaction-separation on continuous chromatographic system[J].Chemical Engineering journal and the biochemical engineering,1992,50:23-28.
[76]Coelho M S,Azevedo D C,Teixeira J A,et al.Dextran and fructose separation on an SMB continuous chromatographic unit[J].Biochemical Engineering Journal,2002,12:215-221.
[77]陈峡华,张亚雯,刘硝.右旋糖酐废液中提取果糖试验[J].淀粉和淀粉糖,1993,(1):15-18.
[78]李玲.多糖的分离、纯化与分析[J].新疆工学院学报,1996,17(2):128-132.
[79]管军军,张春霞.多糖的研究[J].广东饲料,2003,12(3):32-34.
[80]张惟杰.糖复合物生化研究技术[M].浙江:浙江大学出版社,1994:133.
[81]Nilsoon G,Nilsson K.Molecular-weight distribution determination of clinical dextran by gel permeation chromatography[J].Chromatography,1974:101(1):137-153.
[82]Alosp R M,Vlachogiannis G J.Determination of the molecular weight of clinical dextran by gel permeation chromatography on TSK PW type columns[J].Journal of Chromatography,1982,246(2):227-240.
[83]方积年.多糖的分离纯化及其纯度鉴别与分子量测定[J].药学通报,1984,19:46.
[84]魏远安,方积年.高效凝胶渗透色谱法测定多糖纯度及分子量[J].药学学报,1989,24(7):532-536.
[85]盛以虞,仲惠娟.高效凝胶渗透色谱法测定壳聚糖的分子量[J].中国药科大学学报,1994,25(4):242-244.
[86]董慧茹,柯以侃.仪器分析[M].北京:化学工业出版社,2000:190.
[87]孙力军,吴吉明.肠膜明串珠菌的分离与鉴定[J].安徽农业技术师范学院学报,2000,14(4):22-25.
[88]杨汝德.现代工业微生物学教程[M].北京:高等教育出版社,2006:98-101.
[89]Karthikeyan R S,Rakshit S K,Baradarajan A.Optimization of batch condition for dextran production[J].Bioprocess Engineering,1996,15:247-251
[90]Gil E C,Colarte A I,Ghzaoui A E,et al.A sugar cane native dextran as an innovative functional excipient for the development of pharmaceutical tablets[J].European Journal of Pharmaceutics and Biopharmaceutics,2008,68:319-329.
[91]马放,杨基先.环境生物制剂的开发与应用[M].北京:化学工业出版社,2004,148-150.
[92]王洪祚,刘世勇。酶和细胞的固定化[J].化学通报,1997,(2):22-27.
[93]郭杰炎,蔡武城.微生物酶[M].科学出版社,1986:182-184.
[94]饶应福,夏四清,姜剑.固定化微生物技术在环境治理中的应用[J].2005,19(2):24-26.
[95]何延青,刘俊良,杨平,等.微生物固定化技术与载体结构的研究[J].环境科学,2004.25:101-104.
[96]况金蓉.生物吸附技术处理重金属废水的应用[J].武汉理工大学学报.2002,26(3):401-403.
[97]王里奧,崔志强,钱宗琴,等.微生物固定化的发展及在废水处理中的应用[J].重庆大学学报,2004,27(3):126-129.
[98]贺小贤.生物工艺原理[M].北京:化学工业出版社,2003,215-218.
[99]张鸿.用固定化微生物的反应器处理含重金属离子的废水[D].南京:东南大学,2003
[100]王彩冬,黄兵,罗欢.固定化微生物技术及其应用研究进展[J].云南化工,2007,34(4):79-82,91.
[2]张洪斌.海藻酸钠固定化酶制备右旋糖酐的研究[D].安徽:合肥工业大学,2004。
[3]章朝辉.右旋糖酐的制备及应用[J].四川化工与腐蚀控制,2001,4(1):50-52.
[4]Kelly W J,Asmundson R V,Harrison G L.Differentiation of dextran-producing Leuconostoc strains from ferment rice cake(puto)using pulsed-field gel electrophoresis[J].Food Microbiology,1995,26:345-352.
[5]陈代杰,金飞燕.生物高分子(第五卷 多糖Ⅰ-原核生物多糖)[M].北京:化学工业出版社,2004:305-320.
[6]刘卫东,张启祥.人工胶体血浆代用品的分类和临床应用比较[J].中国药业,2006,15(12):62-63.
[7]章朝辉.蔗糖化学衍生物[J].精细石油化工进展,2000,1(9):8-13.
[8]章思规.精细有机化学品技术手册(上册)[M].北京:科学出版社,1992:583-584.
[9]原正平,王汝龙.化工产品手册[M].北京:化学工业出版社,1987:547-548.
[10]国家药典委员会.中华人民共和国药典(二部)[Z].北京:化学工业出版社,2005:88-90.
[11]廖威.利用生物技术防治甘蔗糖厂“蔗饭”问题的基础研究[D].广西:广西大学:2002.
[12]张力田.碳水化合物化学[M].北京:轻工业出版社,1988:377.
[13]王少武,姚日生.右旋糖酐铁的合成新工艺[J].安徽化工,2000,(5):12-14.
[14]高永锐,杨祥开,管忠谊,等.右旋糖酐铁合成工艺改进实验[J].广西畜牧兽医,2003,19(4):151.
[15]宋春和.右旋糖酐铁制备方法探讨[J].实用医技杂志,1994,4(7):526-527.
[16]范慧红,刘金秀,徐康森.高效凝胶渗透色谱在右旋糖酐铁质控中的应用[J].药物分析杂志,2000,20(4):254-257.
[17]王少武,姚日生.右旋糖酐铁合成新工艺[J].安徽化工,2003,(5):12-13.
[18]Seo E S,Lee J H,Park J Y,et al.Enzymatic synthesis and anti-coagulant effect of salicin analogs by using the Leuconostoc mesenteroides glucansucrase acceptor reaction[J].Journal of Biotechnology,2005,117:31-38.
[19]江幼岷,陈仕儒,马恩普,等.造血干细胞动员剂硫酸葡聚糖的毒性及其含硫量的关系[J].实验血液学杂志,1995,3(1):74-77.
[20]Kim D,Robyt J F.Dextransucrase constitutive mutant of Leuconostoc mesenteroides B-1299[J].Enzyme microbial Technology,1995,17:1050-1056.
[21]葛崇涛,田星,高莉莉.肠膜明串株菌生长特性研究[J].中国乳品工业,2005,(7):72-73.
[22]Kim D,Robyt J F.Production and selection of mutant of Leuconostoc mesenteroides constitutive for glucansucrase[J].Enzyme Microbial Technology,1994,16:659-664.
[23]Kim K,Kim D W,Lee J H,et al.Development of constitutive dextransucrase hyper-producing mutants of Leuconstoc mesenteroides using the synchrotron radiation in the 70-1000 eV region[J].Biotechnology Technology,11:319-321.
[24]Kitaoka M,Robyt J F.Use of a microtiter plate screening method for obtaining Leuconostoc mesenteroides mutant constitutive for glucansucrase[J].Enzyme Microbial Technology,1998,22:527-531.
[25]Santos M,Teixeira J,Rodrigues A.Production of dextransucrase,dextran and fructose from sucrose using Leuconostoc Mesenteroides NRRL B512F[J].Biochemical Engineering Journal,2000,4:177-188.
[26]Yoon S H,Fulton D B,Robyt J F.Enzymatic synthesis of two salicin analogues by reaction of salicyl alcohol with Bacillus macerans cyclomaltodextrin glucanyltransferase and Leuconostoc mesenteroides B-742CB dextransucrase[J].Carbohydrate Research,2004,339:1517-1529.
[27]宋明,金中,徐兰,等.乳酸细菌葡聚糖蔗糖酶研究应用与进展[J].检验医学教育,2005,12(2):30-32.
[28]Zivomir P,Suzana R,Dragan P,et al.Immobilization of dextran- sucrase on regenerated benzoyl cellulose carriers[J].Enzyme and Microbial Technology,2007,40:763-768.
[29]Bertrand A,Morel S,Lefoulon F,et al.Leuconostoc mesenteroides glucansucrase synthesis of flavonoid glucosides by acceptor reactions in aqueous-organic solvents[J]. Carbohydrate Research,2006,341:855-863.
[30]Ara'nzazu G S,Miguel A,Manuel B,et al.Synthesis of methyl-d-glucooligos accharidesbyen trapped dextransucrase from Leuconostoc mesenteroides B-1299[J].Journal of Biotechnology,2006,124:439-445.
[31]Monchois V,Willemot R M,Monsan P,et al.Glucansucrases:mechanism of action and structure-function relationships[J].FEMS Microbiology Reviews,1999,23(2):131-151.
[32]邵彦春,王建华,滕达,等.右旋糖酐蔗糖酶基因的克隆及其序列分析[J].微生物学报,2005,32(3):213-23.
[33]Robyt J F,Kimble B K,Walseth T F.The mechanism of dextransucrase action Direction of biosynthesis[J].Biochem.Biophys,1974,165(2):634-640.
[34]廖威.葡聚糖产生菌发酵培养基优化初步研究[J].广西科学院学报,2003,19(3):125-128.
[35]王勇,张文革,刘石.右旋糖酐制取新工艺研究[J].生物技术,1994,4(3):45-46.
[36]崔益清.右旋糖酐生产新工艺-发酵液水解法[J].药物研究,1999,8(6):17.
[37]张洪斌,姚日生,朱慧霞,等.发酵法生产右旋糖酐的工艺研究[J].合肥工业大学学报,2004,783-787.
[38]李清娣,高歌.右旋糖酐工艺与影响质量的因素[J].中国医学工业杂志,1994,25(2):91-92.
[39]井明冉.右旋糖酐生产过程蛋白胨用量探讨[J].石河子科技,2000,(5):28-29.
[40]廖威,杨辉,梁海秋,等.葡聚糖产生菌的分离鉴定与发酵条件的初步研究[J].食品科学,2003,24(11):56-59.
[41]宋学宁,庄丽萍,傅茂东.影响右旋糖酐质量的因素分析[J].山东医药工业,2003,22(6):46-47.
[42]王冶业,刘晓凤,魏甲乾.固定化肠膜明串珠菌生产右旋糖酐的研究[J].甘肃工业大学学报,2003,29(3):79-81.
[43]郭德祥.关于提高右旋糖酐发酵产率的探讨[J].山东医药工业,1994,13(2):22-23.
[44]郭杰炎,蔡武城.微生物酶[M].北京:科学出版社,1986:172-224.
[45]Sinha J,Dey P K,Panda T.Extractive fermentation for improved production of endoglucanase by an intergeneric fusant of Trichoderma reesei/Saccharomyces cerevisiae using aqueous two-pHase system[J].Biochemical Engineering Journal,2000,6:163-175.
[46]Mase T,Matsumiya Y,mori S,et al.Purification and Characterization of a Novel Glucoamylase from Acremonium sp.YT-78[J].Journal of Fermentation and Bioengineering,1996,81(4):347-350.
[47]Ivanova V,Yankov D,Kabaivanova L,et al.Simultaneous biosynthesiS and purification of two extracellular Bacillus hydrolases in aqueous two-phase systems[J].MiCrObiology Research,2001,156:19-30.
[48]Kitaoka M,Robyt J F.Use of a microtiter plate screening method for obtaining Leuconostoc mesenteroides mutants constitutive for glucansucrase[J].Enzyme and Microbial Technology,1998,22:527-531.
[49]Santos M,Rodrigues A,TeiXeira J A.Production of dextran and fructose from carob pod extract and cheese whey by Leuconostoc mesenteroides NRRL B512F[J].Biochemical Engineering Journal,2005,25:1-6.
[50]郭振友,许丹枫,吴明.超滤技术代替乙醇沉淀粗右旋糖酐工艺研究[J].中国纺织大学学报,2000,26(2):100-102.
[51]Ko E A,Nam S H,Jin X J,et al.Synthesis of thermo- and acid-stable novel oligosaccharides by using dextransucrase with high concentration of sucrose[J].Journal of Applied Glycoscience,2007,54(2):147-155.
[52]Christian T J,Merilyn M H,Richards G N.A preliminary,study of the use of larch arabinogalactan in aqueous two-pHase systems[J].Carbohydrate Polymers,1998,35:7-12.
[53]Kelly W J,Asmundson R V,Harrison G L,et al.Differentiation of dextran-producing Leuconostoc strains from fermented rice cake(puto)using pulsed-field gel electrophoresis[J].International Journal of Food Microbiogy,1995,26:345-352.
[54]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and Glucosyltransferase activity[J].Carbohydrate Research,2004,339:1029-1034.
[55]Rodrigues S,Lona M F,Franco T T.Optimizing panose production by modeling and simulation using factorial design and surface response analysis[J].Journal of Food Engineering,2006,75:433-440.
[56]Cote' G L,Sheng S.Penta-,hexa-,and heptasaccharide acceptor products of alternansucrase[J].Carbohydrate Research,2006,341(2):2066-2072.
[57]Kawakita H,Seto H,Ohto K,et al.Pore control with dextran generated from immobilized dextransucrase[J].Biochemical Engineering Journal,2007,36:190-193.
[58]Eom H J,Seo D M,Han N S.Selection of psychrotropHic Leuconostoc spp.producing highly active dextransucrase from lactate fermented vegetables[J].International Journal of Food Microbiology,2007,117:61-67.
[59]Goulas A K,Fisher D A.,Grimble G K.,et al.Synthesis of isomaltooligosaccharides and oligodextrans by the combined use of dextransucrase and dextranase[J].Enzyme and Microbial Technology,2004,35:327-338.
[60]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and glucosyltransferase activity[J].Carbohydrate Research,2004,339:1029-1034.
[61]Richard G,Yu S,Monsan P.A novel family of glucosyl 1,5-anhydro-D- fructose derivatives synthesised by transglucosylation with dextransucrase from Leuconostoc mesenteroides NRRL B-512F[J].Carbohydrate Research,2005,340:395- 401.
[62]Funane K,Ishii A,Ono H,et al.Changes in linkage pattern of glucan products induced by substitution of Lys residues in the dextransucrase[J].FEBS Letters,2005,579:4739-4745.
[63]Kang H K,Seo M Y,Seo E S,et al.Cloning and expression of levansucrase from Leuconostoc mesenteroides B-512 FMC in Escherichia coli[J].Biochimica et BiopHysica Acta,2005,1727:5-15.
[64]杨敏.酶法生产右旋糖酐[P],C12P 19/08.2000,11,02.
[65]姚日生,董明辉,朱慧霞.酶法合成右旋糖酐相对分子质量的控制[J].生物工程,2007,24(7):964-967.
[66]刘汝宽,李昌珠,张玉军.树脂固定脂肪酶技术的研究[J].河南工业大学学报(自然科学版),2007,28(3):68-71.
[67]Berensmeier S,Ergezinger M,Bohnet M,et al.Design of immobilised dextransucrase for fluidised bed application[J].Journal of Biotechnology,2004,114(3):255-267.
[68]Devulapalle K S,Segura A G,Ferrer M,et al.Effect of carbohydrate fatty acid esters on Streptococcus sobrinus and glucosyltransferase activity[J].Carbohydrate Research,2004, 339:1029-1034.
[69]Kubik C,Sikora B,Bielecki S.Immobilization of dextransucrase and its use with soluble dextranase for glucooligosaccharides synthesis[J].Enzyme and Microbial Technology,2004,34:555-560.
[70]Kitaoka M,Robyt J F.Mechanism of the action of Leuconostoc Mesenteroides B-512FMC dextransucrase:kinetics of the transfer of D-glucose to maltose and the effect of enzyme and substrate concentrations[J].Carbohydrate Research,1999,320:183-191.
[71]陈建龙,祁建城,曹仪植.固定化酶研究进展.化学与生物工程[J],2006,23(2):7-9.
[72]袁定重,张秋禹,侯振宇,等.固定化酶载体材料的最新研究进展[J].材料导报,2006,20(1):69-72.
[73]范友华,黄军,陈泽君.固定化酶载体的研究现状及其应用[J].湖南林业科技,2007,34(2):29-31.
[74]Zafar I,Barker P E.An experimental and computational study of a biochemical polymerisation reaction in a chromatographic reactor separator[J].Chemical Engineering Science,1988,43(9):2369-2375.
[75]Barker P E,Ganetsos G.,Ajongwen J,et al.Bioreaction-separation on continuous chromatographic system[J].Chemical Engineering journal and the biochemical engineering,1992,50:23-28.
[76]Coelho M S,Azevedo D C,Teixeira J A,et al.Dextran and fructose separation on an SMB continuous chromatographic unit[J].Biochemical Engineering Journal,2002,12:215-221.
[77]陈峡华,张亚雯,刘硝.右旋糖酐废液中提取果糖试验[J].淀粉和淀粉糖,1993,(1):15-18.
[78]李玲.多糖的分离、纯化与分析[J].新疆工学院学报,1996,17(2):128-132.
[79]管军军,张春霞.多糖的研究[J].广东饲料,2003,12(3):32-34.
[80]张惟杰.糖复合物生化研究技术[M].浙江:浙江大学出版社,1994:133.
[81]Nilsoon G,Nilsson K.Molecular-weight distribution determination of clinical dextran by gel permeation chromatography[J].Chromatography,1974:101(1):137-153.
[82]Alosp R M,Vlachogiannis G J.Determination of the molecular weight of clinical dextran by gel permeation chromatography on TSK PW type columns[J].Journal of Chromatography,1982,246(2):227-240.
[83]方积年.多糖的分离纯化及其纯度鉴别与分子量测定[J].药学通报,1984,19:46.
[84]魏远安,方积年.高效凝胶渗透色谱法测定多糖纯度及分子量[J].药学学报,1989,24(7):532-536.
[85]盛以虞,仲惠娟.高效凝胶渗透色谱法测定壳聚糖的分子量[J].中国药科大学学报,1994,25(4):242-244.
[86]董慧茹,柯以侃.仪器分析[M].北京:化学工业出版社,2000:190.
[87]孙力军,吴吉明.肠膜明串珠菌的分离与鉴定[J].安徽农业技术师范学院学报,2000,14(4):22-25.
[88]杨汝德.现代工业微生物学教程[M].北京:高等教育出版社,2006:98-101.
[89]Karthikeyan R S,Rakshit S K,Baradarajan A.Optimization of batch condition for dextran production[J].Bioprocess Engineering,1996,15:247-251
[90]Gil E C,Colarte A I,Ghzaoui A E,et al.A sugar cane native dextran as an innovative functional excipient for the development of pharmaceutical tablets[J].European Journal of Pharmaceutics and Biopharmaceutics,2008,68:319-329.
[91]马放,杨基先.环境生物制剂的开发与应用[M].北京:化学工业出版社,2004,148-150.
[92]王洪祚,刘世勇。酶和细胞的固定化[J].化学通报,1997,(2):22-27.
[93]郭杰炎,蔡武城.微生物酶[M].科学出版社,1986:182-184.
[94]饶应福,夏四清,姜剑.固定化微生物技术在环境治理中的应用[J].2005,19(2):24-26.
[95]何延青,刘俊良,杨平,等.微生物固定化技术与载体结构的研究[J].环境科学,2004.25:101-104.
[96]况金蓉.生物吸附技术处理重金属废水的应用[J].武汉理工大学学报.2002,26(3):401-403.
[97]王里奧,崔志强,钱宗琴,等.微生物固定化的发展及在废水处理中的应用[J].重庆大学学报,2004,27(3):126-129.
[98]贺小贤.生物工艺原理[M].北京:化学工业出版社,2003,215-218.
[99]张鸿.用固定化微生物的反应器处理含重金属离子的废水[D].南京:东南大学,2003
[100]王彩冬,黄兵,罗欢.固定化微生物技术及其应用研究进展[J].云南化工,2007,34(4):79-82,91.