Water-soluble and water-insoluble glucans produced by Escherichia coli recombinant dextransucrases from Leuconostoc mesenteroides NRRL B-512F
2001
Funane, K. | Ishii, T. | Matsushita, M. | Hori, K. | Mizuno, K. | Takahara, H. | Kitamura, Y. | Kobayashi, M.
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 13C 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.
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