ABSTRACT
Aims: The cosmic production of biomass and bioactive compounds at pilot scale with minimum production costs is an important task to achieve feasible production process of corresponding secondary metabolites at a commercial level. Materials and Methods: The cell suspension cultures of Catharanthus roseus in MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (9.05 µM), kinetin (4.52 µM) were scaled up in a pilot plant bioreactor (100 lit). The cost of production was reduced by addition of substitute carbon source in a basal medium which hardly costs 30% in the medium. Preliminary studies were performed in the 7-lit bioreactor. A 100 lit stainless steel bioreactor equipped with helical impeller top mounted was used for scale-up of C. roseus suspension cultures and ajmalicine production. Results: The culture medium reduced the cost by 36% by addition of commercial grade sugar whereas medium consist of tissue culture grade sucrose costs 53 USD per 100 lit. The suspension cultures were cultivated in a 100 lit bioreactor containing MS medium fortified with cost-effective carbon source produced ajmalicine 73.18 mg/l DW and achieved 36 kg of fresh biomass on day 20. Conclusion: The results of the present finding demonstrated the feasible and cost-effective production process of ajmalicine at pilot scale.
ABSTRACT
ABSTRACT High performance thin layer chromatographic method (HPTLC) has been developed for the quantification of reserpine and ajmalicine in root part of two different population of Rauvolfia serpentina (L.) Benth. ex Kurz and Rauvolfia tetraphylla L., Apocynaceae, collected from Punjab and Uttarakhand. HPTLC of methanolic extract of root containing indole alkaloids, i.e., reserpine and ajmalicine, was performed on TLC Silicagel 60 F254 (10 cm × 10 cm) plates with toluene:ethyl acetate:formic acid (7:2:1), as mobile phase. Quantification of the reserpine and ajmalicine was performed in the absorption–reflection mode at 268 nm. The recovery of reserpine and ajmalicine were 99.3 and 98.7% respectively. The calibration curves were linear for both the reserpine and ajmalicine, in the range of 200–1200 ng. HPTLC densitometry has been performed for the estimation of reserpine and ajmalicine in root part of R. serpentina and R. tetraphylla for the first time. The method is simple, rapid and cost effective and can be used for routine analysis of ajmalicine and reserpine in different Rauvolfia species as well as for quality control of herbal drugs containing Rauvolfia species.
ABSTRACT
Objective: In order to improve the content of terpenoid indole alkaloids (TIAs), orca3/g10h genes were introduced to the hairy roots in Catharanthus roseus. Methods: Bivalent expression vector CAMBIA1304+ +orca3 + g10h was constructed and introduced into Agrobacterium rhizogenes strain and transformed into C. roseus to obtain transgenic hairy roots. RT-qPCR was used to study the transcriptional differences of relative genes referred to the biosynthesis pathway of TIAs. Then HPLC was used to study TIAs content in the transgenic hairy roots of C. roseus, including vinblastine, vincristine, and ajmalicine. Results: The transcriptional level of genes that linked to biosynthesis of TIAs in the transgenic hairy roots of C. roseus, asα, ggpps, g10h, str, tdc, cpr, sgd, and dat, were all expressed higher than those of the nontransgenic roots. HPLC results showed that modified hairy root of C. roseus owned more total TIAs production, 58.23 mg/g, the number was larger than that of common roots in C. roseus as many as 27.5 times. On the other hand, the average content of vinblastine and vincristine was also more than the common roots in C. roseus. Among them, vinblastine content was the most. The number of production got 51.30 mg/g, which was as many as 197.3 times of the common root of C. roseus. Conclusion: Orca3/g10h double-gene transgenic hairy root of C. roseus can increase TIAs content efficiently.
ABSTRACT
Objective To investigate the characteristics of growth, accumulation of indole alkaloid, and the optical concentration of the nutrition components in the Catharanthus roseus mutant cells induced by the colchicine, looking forward to getting the ideal material producing alkaloids which was suited for industrialized cell cultivation. Methods Mutant cells cultivated on the MS medium were harvested according to experiment designs,and the fresh weight was got.The indole alkaloid as extracted by organic solvent. The ajmalicine and catharanthine extracted from the samples were analyzed by RP-HPLC. Results The growth rate was faster and the indole alkaloid accumulation was more when the mutant cells were cultivated generations 30 and the both were sharply decreased at generations 45, while the contents of medical compounds amounted to the peak at the generations 20. The content of medical components and alkaloid accumulation were evidently prompted by the tryptophan added in the media, especially at the concentration of Ca2+ 1 760 and Zn2+ 12.6 mg/L, respectively. Conclusion C.roseus mutant cells probably is an ideal material for industrialized cell cultivation.
ABSTRACT
Object To investigate the effects of fungal elicitors derived from the fungi Fusarium solani and Aspergillum niger on the accumulation of indole alkaloids in Catharanthus roseus calli. Methods The total indole alkaloid was extracted after the calli were treated with fungal elicitors. Then, the determination of ajmalicine and catharanthine was carried out by RP HPLC. Results The two fungal elicitors stimulate the accumulation not only the total indole alkaloid but the ajmalicine and catharanthine. The optimal exposure time of the two fungal elicitors for different kinds of indole alkaloid was investigated. Conclusion The two fungal elicitors have obvious effect on the accumulation of indole alkaloid in C. roseus calli.