Characterization of Camptotheca acuminata 10-hydroxygeraniol oxidoreductase and iridoid synthase and their application in biological preparation of nepetalactol in Escherichia coli featuring NADP+ - NADPH cofactors recycling
| Autor: | Guolin Zhang, Zhan Liu, Chong Qiao, Yinggang Luo, Jing Pang, Wei Li, Annoor Awadasseid |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Iridoid
medicine.drug_class 02 engineering and technology medicine.disease_cause Biochemistry Cofactor 03 medical and health sciences chemistry.chemical_compound Biotransformation Biosynthesis Structural Biology Oxidoreductase medicine Nepetalactol Molecular Biology Escherichia coli 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology General Medicine 021001 nanoscience & nanotechnology Enzyme chemistry biology.protein 0210 nano-technology |
| Zdroj: | International Journal of Biological Macromolecules. 162:1076-1085 |
| ISSN: | 0141-8130 |
| Popis: | Nepetalactol, an iridoid with four chiral carbons, is a crucial component of aphid sex pheromones that have been employed with great success to control the insect-related diseases. Despite of agricultural usage as end products, iridoids are fundamental biosynthetic intermediates for pharmaceutically important monoterpenoid indole alkaloids such as camptothecin (CAM) and vinca alkaloids. Herein we characterized 10-hydroxygeraniol oxidoreductase (10HGO) and iridoid synthase (IS) from Camptotheca acuminata, a CAM-producing plant, and reported their application in biological preparation of nepetalactol. Ca10HGO and CaIS were respectively cloned from C. acuminata, overexpressed in Escherichia coli, and purified to homogeneity. Ca10HGO catalyzes the oxidation of 10-hydroxygeraniol into 10-oxogeranial, in which NADP+ was reduced to NADPH. CaIS catalyzes nepetalactol formation from 10-oxogeranial using NADPH cofactor. The net outcome of the two reactions generate nepetalactol from 10-hydroxygeraniol efficiently, indicating NADP+ - NADPH recycling. Ca10HGO and CaIS were co-overexpressed in E. coli under optimized fermentation conditions to prepare cell-based catalysts that catalyze the conversion of 10-hydroxygeraniol into nepetalactol. The present work shows the enzymatic conversion of 10-hydroxygeraniol into nepetalactol involved in CAM biosynthesis. Co-overexpression of Ca10HGO and CaIS in E. coli is an alternative valuable cell-based biotransformation process with regenerating recycling of NADP+ - NADPH cofactors for nepetalactol preparation. |
| Databáze: | OpenAIRE |
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