Comparison of two hyoscyamine 6β-hydroxylases in engineering scopolamine biosynthesis in root cultures of Scopolia lurida
- State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700 (China)
- Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Key Laboratory of Plant Ecology and Resources Research in Three Gorges Reservoir Region, SWU-TAAHC Medicinal Plant Joint R&D Centre, School of Life Sciences, Southwest University, Chongqing 400715 (China)
- Tobacco Breeding and Biotechnology Research Center, Yunnan Academy of Tobacco Agricultural Sciences, Key Laboratory of Tobacco Biotechnological Breeding, National Tobacco Genetic Engineering Research Center, Kunming 650021 (China)
- College of Pharmaceutical Sciences, Southwest University, Chongqing 400715 (China)
Highlights: • Overexpression of SlH6H or HnH6H significantly promoted the anisodamine production at similar level. • Overexpression of SlH6H or HnH6H markedly enhanced the scopolamine biosynthesis at different level. • HnH6H exhibited better effects on the enhancement of scopolamine biosynthesis than SlH6H. • Overexpression of HnH6H led to the increased production of total tropane alkaloids. Scopolia lurida, a medicinal plant native to the Tibetan Plateau, is among the most effective producers of pharmaceutical tropane alkaloids (TAs). The hyoscyamine 6β-hydroxylase genes of Hyoscyamus niger (HnH6H) and S. lurida (SlH6H) were cloned and respectively overexpressed in hairy root cultures of S. lurida, to compare their effects on promoting the production of TAs, especially the high-value scopolamine. Root cultures with SlH6H/HnH6H overexpression were confirmed by PCR and real-time quantitative PCR, suggesting that the enzymatic steps defined by H6H were strongly elevated at the transcriptional level. Tropane alkaloids, including hyoscyamine, anisodamine and scopolamine, were analyzed by HPLC. Scopolamine and anisodamine contents were remarkably elevated in the root cultures overexpressing SlH6H/HnH6H, whereas that of hyoscyamine was more or less reduced, when compared with those of the control. These results also indicated that SlH6H and HnH6H promoted anisodamine production at similar levels in S. lurida root cultures. More importantly, HnH6H-overexpressing root cultures had more scopolamine in them that did SlH6H-overexpressing root cultures. This study not only provides a feasible way of overexpressing H6H to produce high-value scopolamine in engineered root cultures of S. lurida but also found that HnH6H was better than SlH6H for engineering scopolamine production.
- OSTI ID:
- 23137342
- Journal Information:
- Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 1 Vol. 497; ISSN 0006-291X; ISSN BBRCA9
- Country of Publication:
- United States
- Language:
- English
Similar Records
Effect of gamma cobalt ($gamma$ $sup 60$Co) radiation on the growth and alkaloidal contents of medicinal plants. Part I. Hyoscyamus niger L. (Solanaceae)
Agrobacterium rhizogenes: Transformed root cultures for the study of polyacetylene metabolism and biosynthesis