Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade
| Autor: | Ding Li, Lin Chen, Yanyan Zhang, Zhang Zhiping, Zhiqiang Chen, Mengfei Huang, Yanzhi He, Shuhang Fan, Xinyue Zhang |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Flavonols General Chemical Engineering Arabidopsis medicine.disease_cause 01 natural sciences General Biochemistry Genetics and Molecular Biology Mixed Function Oxygenases 03 medical and health sciences chemistry.chemical_compound Biosynthesis Affinity chromatography 010608 biotechnology medicine Flavonol synthase Escherichia coli 030304 developmental biology Plant Proteins chemistry.chemical_classification Flavonoids 0303 health sciences biology General Immunology and Microbiology Plant Extracts General Neuroscience chemistry Biochemistry Flavanones biology.protein Kaempferol Quercetin Oxidoreductases Flavanone |
| Zdroj: | Journal of visualized experiments : JoVE. (150) |
| ISSN: | 1940-087X |
| Popis: | Flavonols are a major subclass of flavonoids with a variety of biological and pharmacological activities. Here, we provide a method for the in vitro enzymatic synthesis of a flavonol. In this method, Atf3h and Atfls1, two key genes in the biosynthetic pathway of the flavonols, are cloned and overexpressed in Escherichia coli. The recombinant enzymes are purified via an affinity column and then a bienzymatic cascade is established in a specific synthetic buffer. Two flavonols are synthesized in this system as examples and determined by TLC and HPLC/LC/MS analyses. The method displays obvious advantages in the derivation of flavonols over other approaches. It is time- and labor-saving and highly cost-effective. The reaction is easy to be accurately controlled and thus scaled up for mass production. The target product can be purified easily due to the simple components in the system. However, this system is usually restricted to the production of a flavonol from a flavanone. |
| Databáze: | OpenAIRE |
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