Water-soluble and water-insoluble glucans produced by Escherichia coli recombinant dextransucrases from Leuconostoc mesenteroides NRRL B-512F
Water-soluble and water-insoluble glucans produced by Escherichia coli recombinant dextransucrases from Leuconostoc mesenteroides NRRL B-512F
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DOI：
10.1016/s0008-6215(01)00163-x
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发表时间：
2001-08-03
影响因子：
3.1
通讯作者：
Kobayashi, M
中科院分区：
文献类型：
--
作者：
Funane, K;Ishii, T;Kobayashi, M
来自肠膜明串珠菌NRRL B-512 F的两种葡聚糖蔗糖酶基因dsrS和dsrT 5在大肠杆菌中表达，并且重组dsrT 5葡聚糖蔗糖酶显示出产生水不溶性葡聚糖。相反，来自L.肠膜B-512 F是水溶性的。通过C-13 NMR和糖基键组成分析表明，水不溶性葡聚糖含有约50%的6-连接Glcp和40%的3-连接Glcp。“原始”B-512 F菌株被认为已经产生了含有3-连接Glcp的水不溶性葡聚糖。在大肠杆菌中表达的葡聚糖蔗糖酶产生葡聚糖。糖基组成分析表明，大肠杆菌含有4-连接的Glcp。当不具有C-末端重复单元的截短的、水不溶性的、产生葡聚糖的葡聚糖蔗糖酶被添加到水溶性的、产生葡聚糖的葡聚糖蔗糖酶中时，4-连接的Glcp的量增加。在从B-512 F培养物上清液获得的葡聚糖中，在由从B-512 F菌株培养物上清液纯化的葡聚糖蔗糖酶产生的葡聚糖中，以及在临床葡聚糖中也检测到痕量的4-连接Glcp。糖基键组成分析的结果表明，葡聚糖蔗糖酶产生4-连接的Glcp以及6-和3-连接的Glcp。(C)2001年由Elsevier Science Ltd.出版
Two dextransucrase genes, dsrS and dsrT5, from Leuconostoc mesenteroides NRRL B-512F were expressed in Escherichia coli, and recombinant dsrT5 dextransucrase was shown to produce a water-insoluble glucan. In contrast, native dextran from L. mesenteroides B-512F is water-soluble. The water-insoluble glucan was shown by C-13 NMR and glycosyl-linkage composition analysis to contain about 50% 6-linked Glcp and 40% 3-linked Glcp. The 'primitive' B-512F strain is suggested to have produced water-insoluble glucan containing 3-linked Glcp. The glucans produced by dextransucrases expressed in E. coli contained 4-linked Glcp, as shown by glycosyl-linkage composition analysis. The amount of 4-linked Glcp was increased when the truncated, water-insoluble, glucan-producing dextransucrase, which does not have C-terminal repeating units, was added to the water-soluble, glucan-producing dextransucrase. Trace amounts of 4-linked Glcp were also detected in the dextran obtained from the B-512F culture supernatant, in dextran produced by dextransucrase purified from the B-512F strain culture supernatant, and in clinical dextran. The results of glycosyl-linkage composition analysis suggest that dextransucrases produce 4-linked Glcp as well as 6- and 3-linked Glcp. (C) 2001 Published by Elsevier Science Ltd.