Analysis of the Effect of N555 Mutations on the Product Specificity of Dextransucrase Using Caffeic Acid Phenethyl Ester as an Acceptor Substrate
| Autor: | Xiao-qin Yu, Xiao-jie Ding, Lan-hua Liu, Jing-wen Yang, Xue-qin Hu, Hong-bin Zhang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
chemistry.chemical_classification Stereochemistry 010401 analytical chemistry Mutagenesis Mutant Substrate (chemistry) General Chemistry Phenylethyl Alcohol 01 natural sciences 0104 chemical sciences Dextransucrase chemistry.chemical_compound Residue (chemistry) Caffeic Acids Enzyme chemistry Glucosyltransferases Mutation General Agricultural and Biological Sciences Caffeic acid phenethyl ester Site-directed mutagenesis 010606 plant biology & botany |
| Zdroj: | Journal of Agricultural and Food Chemistry. 69:5774-5782 |
| ISSN: | 1520-5118 0021-8561 |
| DOI: | 10.1021/acs.jafc.1c00822 |
| Popis: | Bioglycosylation is an efficient strategy to improve biological activities and physicochemical properties of natural compounds to develop structural modifications of drugs. In this study, an N555 residue was identified as a candidate for site-directed mutagenesis through sequence alignment with GTF180ΔN. Caffeic acid phenethyl ester (CAPE) was used as an acceptor substrate. Two generated mutants, N555Q and N555E, demonstrated significant specificity of distribution of products. Under identical conditions, the conversion rates of diglycoside products (CAPE-2G) generated by the N555E (80.8%) and N555Q (84.5%) mutants were 3.30- and 3.46-fold higher than those generated by the original enzyme (24.4%). The structural simulation results demonstrated that a new hydrogen bond was formed between the N555 residue and CAPE, and the N555 residue was closely related to substrate elongation. These results provide a reference for subsequent studies. Suitable mutants for transfer of diglycosides have important application potential in the food and pharmaceutical industries. |
| Databáze: | OpenAIRE |
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