Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, Iizuka, Japan.
FEBS J. 2011 Feb;278(3):531-40. doi: 10.1111/j.1742-4658.2010.07973.x. Epub 2010 Dec 23.
Glucosyltransferase (GTF)-I from cariogenic Streptococcus sobrinus elongates the α-(1→3)-linked glucose polymer branches on the primer dextran bound to the C-terminal glucan-binding domain. We investigated the GTF-I-catalyzed glucan synthesis reaction in the absence of the primer dextran. The time course of saccharide production during dextran-independent glucan synthesis from sucrose was analyzed. Fructose and glucose were first produced by the sucrose hydrolysis. Leucrose was subsequently produced, followed by insoluble glucan [α-(1→3)-linked glucose polymers] after a lag phase. High levels of intermediate nigerooligosaccharide series accumulation were characteristically not observed during the lag phase. The results from the enzymatic activity of the acceptor reaction for the nigerooligosaccharide with a degree of polymerization of 2-6 and methyl α-D-glucopyranoside as a glucose analog indicate that the activity increased with an increase in the degree of polymerization. The production of insoluble glucan was numerically simulated using the fourth-order Runge-Kutta method with the kinetic parameters estimated from the enzyme assay. The simulated time course provided a profile similar to that of experimental data. These results define the relationship between the kinetic properties of GTF-I and the time course of saccharide production. These results are discussed with respect to a mechanism that underlies efficient glucan synthesis.
致龋变形链球菌的葡糖基转移酶(GTF)-I 可使结合于 C 末端葡聚糖结合域的引物右旋糖酐上的α-(1→3)-连接葡萄糖聚合物支链延长。我们研究了在没有引物右旋糖酐的情况下 GTF-I 催化的葡聚糖合成反应。分析了蔗糖引发的葡聚糖合成过程中糖的产生的时程。蔗糖水解首先产生果糖和葡萄糖,然后在滞后阶段产生莱苏糖,随后产生不溶性葡聚糖[α-(1→3)-连接葡萄糖聚合物]。在滞后阶段,通常不会观察到中间的黑曲霉寡糖系列的积累。用聚合度为 2-6 的黑曲霉寡糖和甲基α-D-吡喃葡萄糖苷作为葡萄糖类似物的受体反应的酶活性结果表明,该活性随聚合度的增加而增加。使用从酶测定中估算的动力学参数,通过四阶龙格-库塔法对不溶性葡聚糖的产生进行数值模拟。模拟的时程提供了与实验数据相似的图谱。这些结果定义了 GTF-I 的动力学特性与糖的产生时程之间的关系。这些结果结合了高效葡聚糖合成的机制进行了讨论。