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Characterization of the Inserted Mutagenesis Dextransucrases From Leuconostoc Mesenteroides 0326 to Produce Hyperbranched Dextran

Characterization of the Inserted Mutagenesis Dextransucrases From Leuconostoc Mesenteroides 0326 to Produce Hyperbranched Dextran

Chao Wang, Shuang Chen, Hong-Bin Zhang, Yao Li, Xue-Qin Hu

Int J Biol Macromol. 2018 Jun;112:584-590.

PMID: 29408211

Abstract:

Dextran produced by dextransucrase hold strong potential for industrial applications. The exact determinants of the linkage specificity of glucansucrase enzymes have remained largely unknown. Previous studies have investigated the relationships between structure and linkage specificity of the dextransucrase DSR from Leuconostoc mesenteroides by the site-directed mutagenesis of the catalytic pocket. The glycosidic linkage of dextran produced by mutant enzymes changed slightly by 3% to 20%. The mutagenesis dextransucrases were constructed by inserting an amino acid into a catalytic pocket to investigate the product specificities of dextransucrase thoroughly. The sequence and structural analysis of glycoside-hydrolase family 70 enzymes led to two sequences (Motif II and Motif IV) being targeted, which were inserted by saturation mutagenesis and simultaneously recombined between A552 and V553, D662, and S663. Variants with catalytic activity were screened of the library, which synthesizes high molecular weight α-glucans with different proportions of α(1-4) linkages, which ranges from 0% to 52%. Mutant sequence analysis, biochemical characterization, and molecular modeling studies revealed the mechanism of product specificities. The mutant dextransucrase, which synthesizes hyperbranched dextran, were obtained by the novel mutagenesis method. The different properties of dextran provide the foundation for subsequent studies and application.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP9004540-A	Dextran from Leuconostoc mesenteroides		9004-54-0	Price

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