O-β-D-xylopyranosyl-(1-4)-O-<α-L-arabinofuranosyl-(1-3)>-O-β-D-xylopyranosyl-(1-4)-D-xylopyranose | 144874-97-5

• 分子结构分类: 有机化合物 - 有机氧化合物
• 英文名称: O-β-D-xylopyranosyl-(1-4)-O-<α-L-arabinofuranosyl-(1-3)>-O-β-D-xylopyranosyl-(1-4)-D-xylopyranose
• 英文别名: O-α-L-Arabinofuranosyl-(1->3)-O-<β-D-xylopyranosyl-(1->4)>-O-β-D-xylopyranosyl-(1->4)-D-xylose；Araf(a1-3)[Xyl(b1-4)]Xyl(b1-4)Xyl；(2S,3R,4S,5R)-2-[(3R,4R,5R,6S)-4-[(2S,3R,4R,5S)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]oxy-5-hydroxy-6-[(3R,4R,5R)-4,5,6-trihydroxyoxan-3-yl]oxyoxan-3-yl]oxyoxane-3,4,5-triol
• CAS: 144874-97-5；144938-90-9
• 化学式: C₂₀H₃₄O₁₇
• 分子量: 546.48
• InChiKey: NZDJRCIXXFUZID-YSJCTKIVSA-N

计算性质

• 辛醇/水分配系数(LogP)：
  -6.6
• 重原子数：
  37
• 可旋转键数：
  7
• 环数：
  4.0
• sp3杂化的碳原子比例：
  1.0
• 拓扑面积：
  267
• 氢给体数：
  10
• 氢受体数：
  17

反应信息

• 作为产物：
  描述：

  xylotriose 、 4-硝基苯基-ALPHA-L-阿拉伯糖甙 在 Thermobacillus xylanyliticus α-L-arabinofuranosidase R69H-G179F-L352M mutant 作用下， 以 重水 为溶剂， 以17.3%的产率得到α-L-arabinofuranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→4)-D-xylopyranose
  参考文献：
  名称：

  Molecular Design of Non-Leloir Furanose-Transferring Enzymes from an α-l-Arabinofuranosidase: A Rationale for the Engineering of Evolved Transglycosylases

  摘要：

  The vast biodiversity of glycoside hydrolases (GHs) constitutes a reservoir of readily available carbohydrate-acting enzymes that employ simple substrates and hold the potential to perform highly stereopecific and regioselective glycosynthetic reactions. However, most GHs preferentially hydrolyze glycosidic bonds and are thus characterized by a hydrolysis/transglycosylation partition in favor of hydrolysis. Unfortunately, current knowledge is insufficient to rationally modify this partition, specifically mutating key molecular determinants to tip the balance toward transglycosylation. In this study, in the absence of precise knowledge concerning the hydrolysis/transglycosylation partition in a hydrolytic GH51 alpha-L-arabinofuranosidase, we describe how an iterative protein engineering approach has been used to create the first "non-Leloir" transarabinofuranosylases. In the first step, random mutagenesis yielded a point mutation (R69H) at a position that is highly conserved in clan GH-A. Characterization of R69H revealed that this enzyme displays high transglycosylation activity but severely reduced (61-fold) activity on pNP-alpha-L-arabinofuranoside. Upon recombination of R69H with other point mutations selected using semirational or in silico approaches, transfer rates close to 100% and transarabinofuranosylation yields of the main (1 2)-linked oligosaccharide product of 80% (vs 11% for the wild-type) were obtained. Combining data presented here with knowledge drawn from the literature, we suggest that the creation of non-Leloir transglycosylases necessarily involves the destabilization of the highly developed transition states that characterize the predominantly hydrolytic exo-acting GHs; this is an efficient way to prevent ubiquitous water molecules from performing the deglycosylation step.

  DOI：

  10.1021/acscatal.5b00949

文献信息

• Molecular Design of Non-Leloir Furanose-Transferring Enzymes from an α-<scp>l</scp>-Arabinofuranosidase: A Rationale for the Engineering of Evolved Transglycosylases
  作者：Bastien Bissaro、Julien Durand、Xevi Biarnés、Antoni Planas、Pierre Monsan、Michael J. O’Donohue、Régis Fauré

  DOI：10.1021/acscatal.5b00949

  日期：2015.8.7

  The vast biodiversity of glycoside hydrolases (GHs) constitutes a reservoir of readily available carbohydrate-acting enzymes that employ simple substrates and hold the potential to perform highly stereopecific and regioselective glycosynthetic reactions. However, most GHs preferentially hydrolyze glycosidic bonds and are thus characterized by a hydrolysis/transglycosylation partition in favor of hydrolysis. Unfortunately, current knowledge is insufficient to rationally modify this partition, specifically mutating key molecular determinants to tip the balance toward transglycosylation. In this study, in the absence of precise knowledge concerning the hydrolysis/transglycosylation partition in a hydrolytic GH51 alpha-L-arabinofuranosidase, we describe how an iterative protein engineering approach has been used to create the first "non-Leloir" transarabinofuranosylases. In the first step, random mutagenesis yielded a point mutation (R69H) at a position that is highly conserved in clan GH-A. Characterization of R69H revealed that this enzyme displays high transglycosylation activity but severely reduced (61-fold) activity on pNP-alpha-L-arabinofuranoside. Upon recombination of R69H with other point mutations selected using semirational or in silico approaches, transfer rates close to 100% and transarabinofuranosylation yields of the main (1 2)-linked oligosaccharide product of 80% (vs 11% for the wild-type) were obtained. Combining data presented here with knowledge drawn from the literature, we suggest that the creation of non-Leloir transglycosylases necessarily involves the destabilization of the highly developed transition states that characterize the predominantly hydrolytic exo-acting GHs; this is an efficient way to prevent ubiquitous water molecules from performing the deglycosylation step.
• Yoshida; Kusakabe; Matsuo, Agricultural and Biological Chemistry, 1990, vol. 54, # 2, p. 449 - 457
  作者：Yoshida、Kusakabe、Matsuo、Shimizu、Yasui、Murakami

  DOI：——

  日期：——