Understanding the transfer reaction network behind the non-processive synthesis of low molecular weight levan catalyzed by Bacillus subtilis levansucrase.
Understanding the transfer reaction network behind the non-processive synthesis of low molecular weight levan catalyzed by Bacillus subtilis levansucrase.
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DOI：
10.1038/s41598-018-32872-7
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发表时间：
2018-10-09
影响因子：
4.6
通讯作者：
Olvera C
中科院分区：
文献类型：
--
作者：
Raga-Carbajal E;López-Munguía A;Alvarez L;Olvera C
枯草芽孢杆菌（Bacillussubtilis，SacB）的果聚糖蔗糖酶在特定的反应条件下，通过大量中间体的非进行性延伸，催化合成低分子量的莱万。为了加深对聚合物延伸机制的理解，我们对莱万合成过程中的低聚果糖谱演变进行了细致的研究。结果表明，在不同的反应阶段，形成了一级和二级中间体系列。通过与使用果聚糖二糖、blastose、1-蔗果三糖、6-蔗果三糖和neo-kestose的受体反应中获得的产物谱进行比较，并通过对一些相关产物的分离和NMR分析，确定了该系列的起源，证明它们都是从蔗糖形成莱万过程中的固有产物。这些结果允许建立非进行性莱万合成中涉及的果糖基转移反应网络。总体而言，我们的结果揭示了在合成的初始步骤中SacB的松弛的受体特异性如何负责形成几个莱万系列，其构成最终的低分子量莱万分布。
Under specific reaction conditions, levansucrase from Bacillus subtilis (SacB) catalyzes the synthesis of a low molecular weight levan through the non-processive elongation of a great number of intermediates. To deepen understanding of the polymer elongation mechanism, we conducted a meticulous examination of the fructooligosaccharide profile evolution during the levan synthesis. As a result, the formation of primary and secondary intermediates series in different reaction stages was observed. The origin of the series was identified through comparison with product profiles obtained in acceptor reactions employing levanbiose, blastose, 1-kestose, 6-kestose, and neo-kestose, and supported with the isolation and NMR analyses of some relevant products, demonstrating that all of them are inherent products during levan formation from sucrose. These results allowed to establish the network of fructosyl transfer reactions involved in the non-processive levan synthesis. Overall, our results reveal how the relaxed acceptor specificity of SacB during the initial steps of the synthesis is responsible for the formation of several levan series, which constitute the final low molecular weight levan distribution.
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影响因子：
4.1
作者：
Homann, Arne;Biedendieck, Rebekka;Seibel, Juergen
通讯作者：
Seibel, Juergen
影响因子：
4.4
作者：
Euzenat, O;Guibert, A;Combes, D
通讯作者：
Combes, D
影响因子：
3.1
作者：
Seibel, Juergen;Moraru, Roxana;Hecht, Hans-Juergen
通讯作者：
Hecht, Hans-Juergen
影响因子：
4.1
作者：
HESTRIN, S;FEINGOLD, DS;AVIGAD, G
通讯作者：
AVIGAD, G
影响因子：
4.4
作者：
Porras-Dominguez, Jaime R.;Avila-Fernandez, Angela;Lopez Munguia, Agustin
通讯作者：
Lopez Munguia, Agustin