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双酶耦合催化法合成特定聚合度β-1,3-葡寡糖研究 预览

Synthesis of β-1,3-glucooligosaccharides by double enzyme coupling catalysis
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摘要 采用蔗糖磷酸化酶和昆布二糖磷酸化酶双酶耦合催化工艺合成β-1,3-葡寡糖。以蔗糖为底物,通过蔗糖磷酸化酶磷酸解蔗糖生成α-D-葡萄糖-1-磷酸(α-D-Glucose-1-phosphate,G1P),再以来自纤细眼虫(Euglena gracilis Z)中的昆布二糖磷酸化酶为β-1,3-葡寡糖合成酶实现G1P所带的葡萄糖单元往引物葡萄糖上的添加,通过控制催化反应时间得到不同聚合度的寡糖。在该策略下,反应0.5 h时耦合催化合成寡糖聚合度在4~6之间;反应4 h时,有7~11糖生成;4 h之后反应趋于平衡,并无更高聚合度的寡糖生成,聚合度稳定在4~11之间。对合成的寡糖进行电喷雾串联质谱(electrospray ionization tandem mass spectrometry,ESI-MS/MS)和氢核磁共振(hydrogen nuclear magnetic resonance,^1H-NMR)分析,可证明产物为β-1,3-葡寡糖。对耦合催化条件进行优化,发现在100 mmol/L的磷酸缓冲液(pH 6.5)、100 mmol/L的蔗糖、50 mmol/L的葡萄糖、5 mmol/L的MgCl2、0.1 U/mL昆布二糖磷酸化酶、0.2 U/mL蔗糖磷酸化酶,40℃条件下反应4 h,葡萄糖的转化率可达39.4%。 With unique physiological function,β-1,3-glucooligosaccharides has been widely used as food additives and health care products. It is hard to achieve β-1,3-glucooligosaccharides with low production cost and definite degree of polymerization. Previous studies showed that sucrose phosphorylase and laminaribiose phosphorylase could be used as coupled catalysts for the production of β-1,3-glucooligosaccharides. Sucrose phosphorylase purchased from SIGMA-ALDRICH can phosphorolysis sucrose into glucose-1-phosphate and laminaribiose phosphorylase extracted from Euglena gracilis Z can realize the affixion of α-D-glucose-1-phosphate onto primer glucose,thus β-1,3-glucooligosaccharides is produced. We found that after reaction for 0. 5 h,the degree of polymerization was 4 to 6. When the reaction time reached 4 h,oligosaccharides with degree of polymerization 7 to 11 occurred. After that,the reaction system remained stable and no oligosaccharides with higher degree of polymerization were produced. To achieve better conversion rate of glucose,we optimized the reaction condition and found that the conversion rate could reached 39. 4% after reaction with 100 mmol/L phosphate buffer( pH 6. 5),100 mmol/L sucrose,50 mmol/L glucose,5 mmol/L MgCl2,0. 1 U/mL laminaribiose phosphorylase and 0. 2 U/mL sucrose phosphorylase at 40 ℃ for 4 h. ESIMS/MS and ^1 H-NMR results of the reaction products showed that obtained products was β-1,3-glucooligosaccharides.
作者 贺海涛 张洪涛 曲娟娟 郭进 郑明仪 蒋芸 詹晓北 HE Hai-tao, ZHANG Hong-tao, QU Juan-juan, GUO Jin, ZHENG Ming-yi, JIANG Yun, ZHAN Xiao-bei (Key Laboratory of Carbohydrate Chemistry and Bioteehnology of Ministry of Education, School of Bioteehnology, Jiangnan University, Wuxi 214122, China)
出处 《食品与发酵工业》 CSCD 北大核心 2018年第5期1-9,共9页 Food and Fermentation Industries
基金 国家自然科学基金面上项目(21778022) 国家自然科学基金青年科学基金(31201384) 国家博士后特别资助(2014T70472) 国家博士后基金(2012M20996) 江苏博士后基金和江苏省资助博士后基金(1301011B)
关键词 β-1 3-葡寡糖 昆布二糖磷酸化酶 蔗糖磷酸化酶 纤细眼虫 氢核磁共振 β-1,3-glucooligosaccharide laminaribiose phosphorylase sucrose phosphoulase Euglena gracilis Z hydrogen nuclear magnetic resonance ( ^1H-NMR)
作者简介 硕士研究生;张洪涛副教授为通讯作者,E-mail:htzhang@jiangnan.edu.cn。;詹晓北教授为通讯作者,E-mail:xbzhan@yahoo.com。
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  • 1刘锋,江涛,任素梅.熊果苷合成研究进展[J].日用化学工业,2004,34(4):242-244. 被引量:13
  • 2赵永平,朱积川,王晓峰.睾酮在阴茎勃起功能障碍中的应用及研究进展[J].中国男科学杂志,2007,21(1):69-72. 被引量:12
  • 3VANDAMME E J, VAN LOO J, Laporte A.D. Dynamics and regulation of sucrose phosphorylase formation in Leuconostoc mesenteroides fermentations[J]. Biotechnol Bioeng, 1987, 29: 8-15. 被引量:1
  • 4RUSSELL RR, MUKASA H, SHIMAMURA A, et al. Streptococcus mutans gtfA gent specifies sucrose phosphorylase [J]. Infect Immun, 1988, 56(10) : 2763-2765. 被引量:1
  • 5SILVERSTEIN R, VOET J, REED D, et al. Purification and mechanism of action of sucrose phosphorylase[J]. J Biol Chem, 1967,242:1338-1346. 被引量:1
  • 6KULLIN B, ABRATT VR, REID SJ. A functional analysis of the Bifidobactetium longum cscA and scrP genes in sucrose utilization[J]. Appl Microbiol Biot, 2006, 72(5) : 975-981. 被引量:1
  • 7VAN DEN BROEK LAM, VAN BOXTEL EL, KIEVIT RP, et al. Physico-chemical and transglucosylation properties of recombinant sucrose phosphorylase from Bitidobacterium adolescentis DSM20083 [J]. Appl Microbiol Biot, 2004, 65(2) : 219-227. 被引量:1
  • 8KIM MJ, KWON T, LEE H J, et al. Cloning and expression of sucrose phosphorylase gene from Bifidobacterium longum in E. coli and characterization of the recombinant enzyme[J]. Bioteehnol Lett, 2003, 25(15) : 1211-1217. 被引量:1
  • 9LEE JH, YOON SH, NAM SH, et al. Molecular cloning ofa gene encoding the sucrose phosphorylase from Leuconostoc mesenteroides B-1149 and the expression in Escherichia coli[J]. Enzyme Microb Tech, 2006, 39: 612-620. 被引量:1
  • 10KOGA T, NAKAMURA K, SHIROKANE Y, et al. Purification and some properties of sucrose phosphorylase from Leuconostoc mesenteroides[J]. Agr Biol Chem, 1991, 55(7) : 1805-1810. 被引量:1

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