A Repertoire of Alginate Lyases in the Alginate Polysaccharide Utilization Loci of Marine Bacterium Wenyingzhuangia Fucanilytica：Biochemical Properties and Action Pattern

Li, Jiajing; Pei, Xiaojie; Xue, Changhu; Chang, yaoguang; Shen, Jingjing; Zhang, Yuying

doi:10.2139/ssrn.4384840

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“…The detailed procedures for the expression and purification were consistent with our previous study. 35 2.4. Biochemical Characterization.…”

Section: Gene Cloning and Protein Expressionmentioning

confidence: 99%

Action Pattern of a Novel G-Specific Alginate Lyase: Determination of Subsite Specificity by HPAEC–PAD/MS

Li,

Xue,

Shen

et al. 2023

J. Agric. Food Chem.

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G-specific alginate lyases are important tools for alginate fragment biodegradation and oligosaccharide production, which have great potential in alginate refining research. In this research, a novel G-specific alginate lyase Aly7Ce was cloned, expressed, and characterized, with the optimal reaction conditions at 30 °C and pH 8.0. By employing the UPSEC−VWD−MS method, Aly7Ce was confirmed as a random endoacting alginate lyase. Its minimum substrate was tetrasaccharide, and the final product majorly consisted of disaccharide to tetrasaccharide. HPAEC−PAD/MS method was employed to investigate the structurally different unsaturated alginate oligosaccharides. The substrate recognition and subsite specificity of Aly7Ce were revealed by detecting the oligosaccharide pattern in the enzymatic products with oligosaccharides or polysaccharides as substrates. Aly7Ce mainly attacked the second glycosidic linkage from the nonreducing end of oligosaccharide substrates. The subsite specificity of Aly7Ce was revealed as −2 (M/G), − 1 (G), + 1 (M/G), and +2 (M/G). The regular oligosaccharide products of Aly7Ce could be applied for the efficient preparation of ΔG, ΔGG, and ΔGGG with high purity. The G-specific alginate lyase Aly7Ce with a welldefined product composition and action pattern provided a novel tool for the modification and structural elucidation of alginate, as well as for the targeted preparation of oligosaccharides.

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“…The detailed procedures for the expression and purification were consistent with our previous study. 35 2.4. Biochemical Characterization.…”

Section: Gene Cloning and Protein Expressionmentioning

confidence: 99%

Action Pattern of a Novel G-Specific Alginate Lyase: Determination of Subsite Specificity by HPAEC–PAD/MS

Li,

Xue,

Shen

et al. 2023

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

A Repertoire of Alginate Lyases in the Alginate Polysaccharide Utilization Loci of Marine Bacterium Wenyingzhuangia Fucanilytica：Biochemical Properties and Action Pattern

Cited by 1 publication

References 0 publications

Action Pattern of a Novel G-Specific Alginate Lyase: Determination of Subsite Specificity by HPAEC–PAD/MS

Action Pattern of a Novel G-Specific Alginate Lyase: Determination of Subsite Specificity by HPAEC–PAD/MS

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