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Detailed Structural Characterization of Glucans Produced by Glucansucrases from Leuconostoc citreum TMW 2.1194.
J Agric Food Chem 2019; 67(24):6856-6866JA

Abstract

The water kefir organism Leuconostoc citreum TMW 2.1194 forms highly branched dextrans with O3- and O4-bound side chains. To obtain detailed information on the enzymatic synthesis of these polymers, the four glucansucrases encoded by Leuconostoc citreum TMW 2.1194 were cloned, heterologously expressed, and used for polysaccharide production. Molecular and macromolecular structure of the synthesized glucans were analyzed by methylation analysis, two-dimensional NMR spectroscopy, oligosaccharide analysis after partial hydrolysis, and asymmetric flow field-flow fractionation. It was demonstrated that two glucansucrases form insoluble glucans with variously branched dextran sections and varying portions of consecutive, 1,3-linked glucose units. In contrast, the other two glucansucrases synthesized O3- (Lc6255) and O4-branched (Lc1785) soluble dextrans. Analysis, isolation, and characterization of enzymatically liberated oligosaccharides showed that monomeric and elongated side chains are abundant in both polysaccharides. From the structures and size distributions it was concluded that Lc1785 is mainly responsible for synthesis of fermentatively produced soluble dextrans.

Authors+Show Affiliations

Department of Food Chemistry and Phytochemistry, Institute of Applied Biosciences , Karlsruhe Institute of Technology (KIT) , Karlsruhe 76131 , Germany.Lehrstuhl für Technische Mikrobiologie , Technische Universität München (TUM) , Freising 80333 , Germany.Lehrstuhl für Technische Mikrobiologie , Technische Universität München (TUM) , Freising 80333 , Germany.Department of Food Chemistry and Phytochemistry, Institute of Applied Biosciences , Karlsruhe Institute of Technology (KIT) , Karlsruhe 76131 , Germany.Department of Food Chemistry and Phytochemistry, Institute of Applied Biosciences , Karlsruhe Institute of Technology (KIT) , Karlsruhe 76131 , Germany.Lehrstuhl für Technische Mikrobiologie , Technische Universität München (TUM) , Freising 80333 , Germany.Department of Food Chemistry and Phytochemistry, Institute of Applied Biosciences , Karlsruhe Institute of Technology (KIT) , Karlsruhe 76131 , Germany.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31124355

Citation

Münkel, Franziska, et al. "Detailed Structural Characterization of Glucans Produced By Glucansucrases From Leuconostoc Citreum TMW 2.1194." Journal of Agricultural and Food Chemistry, vol. 67, no. 24, 2019, pp. 6856-6866.
Münkel F, Bechtner J, Eckel V, et al. Detailed Structural Characterization of Glucans Produced by Glucansucrases from Leuconostoc citreum TMW 2.1194. J Agric Food Chem. 2019;67(24):6856-6866.
Münkel, F., Bechtner, J., Eckel, V., Fischer, A., Herbi, F., Jakob, F., & Wefers, D. (2019). Detailed Structural Characterization of Glucans Produced by Glucansucrases from Leuconostoc citreum TMW 2.1194. Journal of Agricultural and Food Chemistry, 67(24), pp. 6856-6866. doi:10.1021/acs.jafc.9b01822.
Münkel F, et al. Detailed Structural Characterization of Glucans Produced By Glucansucrases From Leuconostoc Citreum TMW 2.1194. J Agric Food Chem. 2019 Jun 19;67(24):6856-6866. PubMed PMID: 31124355.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detailed Structural Characterization of Glucans Produced by Glucansucrases from Leuconostoc citreum TMW 2.1194. AU - Münkel,Franziska, AU - Bechtner,Julia, AU - Eckel,Viktor, AU - Fischer,Anja, AU - Herbi,Frauke, AU - Jakob,Frank, AU - Wefers,Daniel, Y1 - 2019/06/07/ PY - 2019/5/28/pubmed PY - 2019/7/19/medline PY - 2019/5/25/entrez KW - HPAEC-PAD KW - NMR spectroscopy KW - dextran KW - enzymatic fingerprinting KW - fermentation KW - mutan KW - oligosaccharides KW - purification SP - 6856 EP - 6866 JF - Journal of agricultural and food chemistry JO - J. Agric. Food Chem. VL - 67 IS - 24 N2 - The water kefir organism Leuconostoc citreum TMW 2.1194 forms highly branched dextrans with O3- and O4-bound side chains. To obtain detailed information on the enzymatic synthesis of these polymers, the four glucansucrases encoded by Leuconostoc citreum TMW 2.1194 were cloned, heterologously expressed, and used for polysaccharide production. Molecular and macromolecular structure of the synthesized glucans were analyzed by methylation analysis, two-dimensional NMR spectroscopy, oligosaccharide analysis after partial hydrolysis, and asymmetric flow field-flow fractionation. It was demonstrated that two glucansucrases form insoluble glucans with variously branched dextran sections and varying portions of consecutive, 1,3-linked glucose units. In contrast, the other two glucansucrases synthesized O3- (Lc6255) and O4-branched (Lc1785) soluble dextrans. Analysis, isolation, and characterization of enzymatically liberated oligosaccharides showed that monomeric and elongated side chains are abundant in both polysaccharides. From the structures and size distributions it was concluded that Lc1785 is mainly responsible for synthesis of fermentatively produced soluble dextrans. SN - 1520-5118 UR - https://www.unboundmedicine.com/medline/citation/31124355/Detailed_Structural_Characterization_of_Glucans_Produced_by_Glucansucrases_from_Leuconostoc_citreum_TMW_2.1194 L2 - https://dx.doi.org/10.1021/acs.jafc.9b01822 DB - PRIME DP - Unbound Medicine ER -