Enzymatic Biosynthesis of Vomilenine, a Key Intermediate of the Ajmaline Pathway, Catalyzed by a Novel Cytochrome P 450-Dependent Enzyme from Plant Cell Cultures of Rauwolfia serpentina

1995 ◽  
Vol 50 (1-2) ◽  
pp. 45-53 ◽  
Author(s):  
Heike Falkenhagen ◽  
Joachim Stöckigt
Keyword(s):  
Cytochrome P450 ◽  
Cell Cultures ◽  
Indole Alkaloid ◽  
Cell Suspension Cultures ◽  
Significant Loss ◽  
Side Reactions ◽  
The Novel ◽  
Ph Optimum ◽  
Rauwolfia Serpentina ◽  
Km Value

Abstract Microsomal preparations from Rauwolfia serpentina Benth. cell suspension cultures cata­lyze a key step in the biosynthesis of ajmaline -the enzymatic hydroxylation of the indole alkaloid vinorine at the allylic C-21 resulting in vomilenine. Vomilenine is an important branch-point intermediate, leading not only to ajmaline but also to several side reactions of the biosynthetic pathway to ajmaline. The investigation of the taxonomical distribution of the enzyme indicated that vinorine hydroxylase is exclusively present in ajmaline-producing plant cells. The novel enzyme is strictly dependent on NADPH2 and O2 and can be inhibited by typical cytochrome P450 inhibitors such as cytochrome c, ketoconazole and carbon mon­oxide (the effect of CO is reversible with light of 450 nm). This suggests that vinorine hy­droxylase is a cytochrome P450-dependent monooxygenase. A pH optimum of 8.3 and a temperature optimum of 40 °C were found. The Km value was 3 μᴍ for NADPH2 and 26 μᴍ for vinorine. The optimum enzyme activity could be determined at day 4 after inoculation of the cell cultures in AP I medium. Vinorine hydroxylase could be stored with 20% sucrose at -28 °C without significant loss of activity.

Structure and Synthesis of a New Indole Alkaloid, 19 (S)-Hydroxy-Nb-methylraumacline, Obtained by the Biotransformation of Ajmaline in Plant Cell Cultures of Rauwolfia Serpentina Benth.

Tetrahedron ◽  
1992 ◽  
Vol 48 (13) ◽  
pp. 2627-2634 ◽  
Author(s):  
Hiromitsu Takayama ◽  
Mariko Kitajima ◽  
Shinya Suda ◽  
Norio Aimi ◽  
Shin-ichiro Sakai ◽  
...  
Keyword(s):  
Plant Cell ◽  
Cell Cultures ◽  
Indole Alkaloid ◽  
Plant Cell Cultures ◽  
Rauwolfia Serpentina

Novel Glucoalkaloids from Rauwolfia Cell Cultures – Acetylrauglucine and Related Glucosides

1988 ◽  
Vol 43 (7-8) ◽  
pp. 479-484 ◽  
Author(s):  
Carl Michael Ruyter ◽  
Helmut Schübel ◽  
Joachim Stöckigt
Keyword(s):  
Cell Suspension ◽  
Cell Cultures ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Production Medium ◽  
Rauwolfia Serpentina

From cell suspension cultures of Rauwolfia serpentina grown in an optimized production medium for the glucoalkaloid raucaffricine, a novel glucoalkaloid was isolated and identified as 17-O-acetyl-21-O-β-ᴅ-glucopyranosyl-ajmaline (acetylrauglucine). This alkaloid is formed in very small amounts (< 5 × 10-4%). The biogenetically related Nα-demethylated base (acetyl-nor-rauglucine) and the deacetyl product rauglucine have also been detected in culture extracts. In addition 21(.R)-(β-D-glucopyranosyl)-hydroxy-sarpagan-17-al has been isolated and identified as an artifact which originates from raucaffricine.

scholarly journals Efficient production of vindoline from tabersonine by metabolically engineered Saccharomyces cerevisiae

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Tengfei Liu ◽  
Ying Huang ◽  
Lihong Jiang ◽  
Chang Dong ◽  
Yuanwei Gou ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
De Novo ◽  
Indole Alkaloid ◽  
Functional Expression ◽  
High Yield ◽  
Efficient Production ◽  
Cytochrome P450 Enzymes ◽  
Copy Numbers ◽  
Yield Production ◽  
Engineered Saccharomyces Cerevisiae

AbstractVindoline is a plant derived monoterpene indole alkaloid (MIA) with potential therapeutic applications and more importantly serves as the precursor to vinblastine and vincristine. To obtain a yeast strain for high yield production of vindoline from tabersonine, multiple metabolic engineering strategies were employed via the CRISPR/Cas9 mediated multiplex genome integration technology in the present study. Through increasing and tuning the copy numbers of the pathway genes, pairing cytochrome P450 enzymes (CYPs) with appropriate cytochrome P450 reductases (CPRs), engineering the microenvironment for functional expression of CYPs, enhancing cofactor supply, and optimizing fermentation conditions, the production of vindoline was increased to a final titer as high as ∼16.5 mg/L, which is more than 3,800,000-fold higher than the parent strain and the highest tabersonine to vindoline conversion yield ever reported. This work represents a key step of the engineering efforts to establish de novo biosynthetic pathways for vindoline, vinblastine, and vincristine.

scholarly journals Oxidative burst and cell death induced by b-quercinin and zoospores of Phytophthora quercina in tobacco cell cultures

2017 ◽  
Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽  
pp. 446-448 ◽  
Author(s):  
J. Koehl ◽  
E.F. Elstner ◽  
W. Oßwald ◽  
I. Heiser
Keyword(s):  
Cell Death ◽  
Cell Suspension ◽  
Cell Cultures ◽  
Oxidative Burst ◽  
Mode Of Action ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Tobacco Cell ◽  
Tobacco Cell Suspension

Mode of action of β-quercinin, a novel elicitin on tobacco cell suspension cultures (cvs. Bel B and Bel W3) was investigated by measuring the oxidative burst and cell death in these cell cultures. β-quercinin induced an oxidative burst comparable to that excited by zoospores from P. quercina. Adding superoxidedismutase, catalase and diphenyleneiodonium to elicited cell cultures, it could be demonstrated, that the induction of cell death in tobacco cell cultures is not correlated to the oxidative burst.

Effects of Plant Bioregulators on the Production of Iridoid Derived Terpenoids in Valeriana wallichii and Fedia cornucopiae Cell Suspension Cultures

1987 ◽  
Vol 42 (1-2) ◽  
pp. 33-40 ◽  
Author(s):  
Wolfram Förster ◽  
Hans Becker
Keyword(s):  
Cell Suspension ◽  
Cell Cultures ◽  
Exponential Growth ◽  
Cell Suspension Cultures ◽  
Growth Cycle ◽  
Suspension Cultures ◽  
Tissue Cultures ◽  
Cell Systems ◽  
Optimal Activity ◽  
Plant Tissue Cultures

Abstract Four plant bioregulators were tested for their effects on production of valepotriates in Valeriana wallichii and Fedia cornucopiae cell suspension cultures. Concentrations of more than 10 ppm reduced valepotriate yield. At lower concentrations production was increased. For optimal activity, bioregulators had to be applied during early exponential growth, up to day 8 of the growth cycle. At equimolar concentrations dim ethylm orpholinium bromide (4 ppm) and dimethylpiperidinium chloride (3 ppm) significantly im proved total valepotriates in V. wallichii (up to 23%) and in F cornucupiae (up to 50% ) 2-(3,4-dichlorophenoxy ) - triethylamine (6 ppm ) and 2-(3,5-diisopropylphenoxy)-triethylam ine (6.4 ppm) increased valepotriate production in both cell cultures up to 40%. With dimethylpiperidinium chloride and dimethylmorpholinium bromide the ratio of m onoene to diene valepotriates in both cell systems was significantly shifted to the m onoene com pounds. A general use of these bioregulators to increase production of terpenoid secondary m etabolites in plant tissue cultures is indicated.

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