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糖苷合成酶——一类新型的寡糖高效合成工具 被引量:7

Glycosynthases: a Novel Efficient Synthetic Tool for Oligosaccharides
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摘要 寡糖是哺乳动物细胞表面糖蛋白和糖脂以及微生物来源的生理活性物质的要素之一,其应用于医药的巨大潜能至今还没有得到充分体现,主要原因是合成足够于临床使用的寡糖非常困难.传统的化学法和酶法在大规模合成寡糖方面都有一定局限性.近年来,分子生物学技术大大推动了糖苷酶合成寡糖的研究,将糖苷酶催化中心亲核体氨基酸定点突变为非亲核体氨基酸,导致酶的原有水解活性丧失,只催化糖苷键合成反应,寡糖产量最高可达99%,人工产生了一类新酶——糖苷合成酶(glycosynthases),随后又产生了硫代糖苷酶(thioglycoligases)和硫代糖苷合成酶(thioglycosynthases).糖苷合成酶的高通量筛选可用双质粒系统和酵母三杂交系统进行,其活性的进一步改进可通过亲核体氨基酸位点不同氨基酸取代、其他位点氨基酸突变、反应条件优化等方法进行,其区域选择性的改变或增强可通过改变糖基受体分子达到.糖苷合成酶作为一种新型高效的生物催化剂,对寡糖的工业化合成有着重要意义,它的出现对糖生物学的发展必将起到巨大的推动作用. Oligosaccharides are one of the essential physiological constituents of glycoproteins and glycolipids on mammalian cell surfaces and microbial metabolites. They have considerable potential as therapeutics but are only now slowly assuming this important role. One of the reasons for their slow development has been the considerable difficulty in synthesizing oligosaccharides on the scale necessary for their clinical evaluation. Classical chemical and enzymatic methods both have limitations in synthesizing large-scale oligosaccharides. In recent years, the rapid progress on molecular biotechnology has promoted the development of retaining glycosidases in oligosaccharides synthesis, which led to the production of a novel class of enzymatic activities termed the glycosynthases. These new enzymes are retaining glycosidase mutants in which the catalytic nucleophile has been converted to a non-nucleophilic residue, synthesizing oligosaccharides in high yields ( the highest yields reach 99%) without any hydrolysis. Furthermore thioglycoligases and thioglycosynthases have been developed subsequently in the past three years. Glycosynthases can be screened in high-throughput assay by the two-plasmid system and the yeast three-hybid system respectively. Their activity can be significantly enhanced by substituting alternative residues for nucleophile, additional random mutations and optimizing reaction conditions. Their regioselectivity can be modified through changes in receptors.
出处 《生物化学与生物物理进展》 SCIE CAS CSCD 北大核心 2006年第4期310-320,共11页 Progress In Biochemistry and Biophysics
基金 国家自然科学基金资助项目(30170008) 国家"十五"攻关计划项目(2004BA713B04-06).
关键词 寡糖合成 糖苷酶 糖苷合成酶 高通量筛选 性质改进 oligosaccharides synthesis, glycosidases, glycosynthases, screening in high-throughput assay,improvement of characteristics
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参考文献37

  • 1Palcic M M.Biocatalytic synthesis of oligosaccharides.Curr Opin Biotechnol,1999,10 (6):616~624 被引量:1
  • 2ZopfD,Roth S.Oligosaccharide anti-infective agents.Lancet,1996,347 (9007):1017~1021 被引量:1
  • 3Crout D H,Vic G.Glycosidases and glycosyl transferases in glycoside and oligosaccharide synthesis.Curr Opin Chem Biol,1998,2 (1):98~ 111 被引量:1
  • 4Sears P,Wong C H.Toward automated synthesis ofoligosaccharides and glycoproteins.Science,2001,291(5512):2344~2350 被引量:1
  • 5Scigelova M,Singh S,Crout D H G.Glycosidases-a great synthetic tool.J Mol Catal B:Enzym,1999,6 (5):483~494 被引量:1
  • 6Daines A M,Maltman B A,Flitsch S L.Synthesis and modifications of carbohydrates,using biotransformations.Curr Opin Chem Biol,2004,8 (2):106~ 113 被引量:1
  • 7McCarter J D,Withers S G.Mechanisms of enzymatic glycoside hydrolysis.Curr Opin Struct Biol,1994,4 (6):885~892 被引量:1
  • 8Rye C S,Withers S G.Glycosidase mechanisms.Curr Opin Chem Biol,2000,4 (5):573~580 被引量:1
  • 9Mackenzie L F,Wang Q,Warren R A J,et al.Glycosynthases:mutant glycosidases for oligosaccharide synthesis.J Am Chem Soc,1998,120 (22):5583~5584 被引量:1
  • 10Malet C,Planas A.From β-glucanase to β-glucansynthase:glycosyl transfer to α-glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile.FEBS Lett,1998,440 (1 ~2):208~212 被引量:1

二级参考文献36

  • 1Ichikawa Y, Look GC, Wong CH. Enzyme-catalyzed oligosaccharide synthesis. Anal Biochem, 1992, 202: 215 - 238. 被引量:1
  • 2He XY, Zhang SZ, Yang SJ. Cloning and expression of a thermostable β-glycosidase Gene from thermus nonproteolyticus HG 102 and haracterization of the recombinant enzyme. Applied Biochemistry and Biotechnology, 2002,94(3): 243- 255. 被引量:1
  • 3Wang XQ, He XY, Yang SJ et al. Structural basis for thermostability of beta-glycosidase from the thermophilic enbacterium thermos nonproteolyticus HG102. Journal of Bacteriology, 2003, 185(14):4248 - 4255. 被引量:1
  • 4Hocker B, Jürgens C, Wilmanns M et al. Stability, catalytic versatility and evolution of the (βα) 8-barrel fold. Current Opinion in Biotechnology, 2001, 12: 376 - 381. 被引量:1
  • 5Watt GM, Lowden PAS, Flitsch SL. Enzyme-catalyzed formation of glycosidic linkages. Curr Opin Struct Biol, 1997, 7: 652- 660. 被引量:1
  • 6Elling L. Glycobiotechnology: enzymes for the synthesis of nucleotide sugars. Adv Biochem Eng Biotechnol, 1997, 58:89 - 144. 被引量:1
  • 7Mackenzie LF, Wang Q, Warren RAJ et al. Glycosynthases-mutant glycosidases for oligosaccharide synthesis. J Am Chem Soc, 1998,120:5583 - 5584. 被引量:1
  • 8Schwede T, Kopp J, Guex N et al. SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Research, 2003,31:3381 - 3385. 被引量:1
  • 9Guex N, Peitsch MC. SWISS-MODEL and the Swiss-Pdb Viewer:An environment for comparative protein modeling. Electrophoresis,1997, 18: 2714- 2723. 被引量:1
  • 10Peitsch MC. Protein modeling by E-mail. Bio/Technology, 1995,13:658-660. 被引量:1

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