| Title: | 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 |
| Author(s): | Awadasseid A; Li W; Liu Z; Qiao C; Pang J; Zhang G; Luo Y; |
| Address: | "Center for Natural Products Research, Chengdu Institute of Biology, Chinese Academy of Sciences, 9 Section 4, Renmin Road South, Chengdu 610041, People's Republic of China; University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049, People's Republic of China. Center for Natural Products Research, Chengdu Institute of Biology, Chinese Academy of Sciences, 9 Section 4, Renmin Road South, Chengdu 610041, People's Republic of China. Center for Natural Products Research, Chengdu Institute of Biology, Chinese Academy of Sciences, 9 Section 4, Renmin Road South, Chengdu 610041, People's Republic of China; State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, People's Republic of China. Electronic address: yinggluo@cib.ac.cn" |
| DOI: | 10.1016/j.ijbiomac.2020.06.223 |
| ISSN/ISBN: | 1879-0003 (Electronic) 0141-8130 (Linking) |
| Abstract: | "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" |
| Keywords: | "Bridged Bicyclo Compounds, Heterocyclic/*metabolism Camptotheca/enzymology/*genetics *Escherichia coli/genetics/metabolism NADP/*metabolism *Oxidoreductases/biosynthesis/genetics *Plant Proteins/biosynthesis/genetics 10-Hydroxygeraniol oxidoreductase Camp;" |
| Notes: | "MedlineAwadasseid, Annoor Li, Wei Liu, Zhan Qiao, Chong Pang, Jing Zhang, Guolin Luo, Yinggang eng Netherlands 2020/07/01 Int J Biol Macromol. 2020 Nov 1; 162:1076-1085. doi: 10.1016/j.ijbiomac.2020.06.223. Epub 2020 Jun 26" |
