Shikonin Production by Callus Culture of Onosma bulbotrichom as Active Pharmaceutical Ingredient (original) (raw)
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Reviews in Analytical Chemistry, 2011
The genus Onosma is represented by 97 species, 4 varieties, 1 hybrid species in the fl ora of Turkey. Some Onosma species with roots containing naphthoquinones (alkannin, shikonin and alkannin/shikonin derivatives) are known as " Havaciva " in Turkey. Onosma nigricaule is an endemic species for Turkey. The roots of O. nigricaule are used for wounds and burns in Erzurum (Turkey). In this study, quantitative determination of shikonin derivatives (deoxyshikonin, β ,β-dimethylacrylshikonin and acetyl shikonin) isolated from the roots of O. nigricaule was performed using a UV-Vis spectrophotometric method. The isolated shikonin derivatives have shown three maxima at 493 nm, 523 nm and 562 nm. The mean absorption coeffi cient of the three shikonin derivatives (deoxyshikonin, β , β-dimethylacrylshikonin and acetyl shikonin) was determined at 523 nm and 562 nm as 15.27 l/g cm and 9.39 l/g cm, respectively. The total content of shikonin derivatives were determined as 214.6 mg/100 g root at 523 nm and 220.6 mg/100 g root at 562 nm by using mean absorption coeffi cient of the three shikonin derivatives (deoxyshikonin, β,β-dimethylacrylshikonin and acetyl shikonin).
Cell Biology International, 2011
Cell suspension cultures of Arnebia euchroma were raised from in vitro leaf-derived friable callus on liquid MS [Murashige and Skoog] medium supplemented with BAP (6-benzylaminopurine) (10.0 mM) and IBA (indole-3-butyric acid) (5.0 mM). A two-stage culture system was employed using growth and production medium for cell biomass and shikonin derivatives, respectively. Factors such as light, temperature, sucrose and pH (hydrogen ion concentration) were studied to observe their effect on the shikonin derivative production. Light conditions completely inhibited shikonin derivative production. Out of different temperature regimes tested, the highest yield (586.17 mg/g FW) was found at 25uC. Maximum production (656.14 mg/g FW) was observed in 6% sucrose. An alkaline pH (7.25-9.50) favoured shikonin derivative production. The results showed that physical and chemical factors greatly influence the production of shikonin derivatives in cell suspension cultures of A. euchroma. Therefore, by employing optimum culture conditions, it is possible to enhance the production of secondary compounds from the cells. The factors optimized for in vitro production of shikonin derivatives during the present study can successfully be employed for their large-scale production in bioreactors.
Biotechnological approaches to the production of shikonins: a critical review with recent updates
Critical Reviews in Biotechnology, 2014
Shikonins are commercially important secondary compounds, known for array of biological activities such as antimicrobial, insecticidal, antitumor, antioxidants, etc. These compounds are usually colored and therefore have application in food, textiles and cosmetics. Shikonin and its derivatives, which are commercially most important of the naphthoquinone pigments, are distributed among members of the family Boraginaceae. These include different species of Lithospermum, Arnebia, Alkanna, Anchusa, Echium and Onosma. The growing demand for plantbased natural products has made this group of compounds one of the enthralling targets for their in vitro production. The aim of this review is to highlight the recent progress in production of shikonins by various biotechnological means. Different methods of increasing the levels of shikonins in plant cells such as selection of cell lines, optimization of culture conditions, elicitation, in situ product removal, genetic transformation and metabolic engineering are discussed. The experience of different researchers working worldwide on this aspect is also considered. Further, to meet market demand, the needs for continuous and reliable production systems, as well as future prospects, are included.
Cell Biology International, 2011
Cell suspension cultures of Arnebia euchroma were raised from in vitro leaf-derived friable callus on liquid MS [Murashige and Skoog] medium supplemented with BAP (6-benzylaminopurine) (10.0 mM) and IBA (indole-3-butyric acid) (5.0 mM). A two-stage culture system was employed using growth and production medium for cell biomass and shikonin derivatives, respectively. Factors such as light, temperature, sucrose and pH (hydrogen ion concentration) were studied to observe their effect on the shikonin derivative production. Light conditions completely inhibited shikonin derivative production. Out of different temperature regimes tested, the highest yield (586.17 mg/g FW) was found at 25uC. Maximum production (656.14 mg/g FW) was observed in 6% sucrose. An alkaline pH (7.25-9.50) favoured shikonin derivative production. The results showed that physical and chemical factors greatly influence the production of shikonin derivatives in cell suspension cultures of A. euchroma. Therefore, by employing optimum culture conditions, it is possible to enhance the production of secondary compounds from the cells. The factors optimized for in vitro production of shikonin derivatives during the present study can successfully be employed for their large-scale production in bioreactors.
Journal of …, 2008
Alkannin/shikonin mixture from roots of Onosma echioides (L.) L.: Extraction method study and quantification This work reports the extraction procedures of alkannin/shikonin mixture from roots of six populations of Onosma echioides, by means of three extraction techniques: Soxhlet extraction, maceration and rapid solid -liquid dynamic extraction (RSLDE). Five solvents with different polarity (hexane, petroleum ether, chloroform, ethyl acetate, methanol) were also studied. Analysis of the extracts was performed by an HPLC-DAD (diode array detector) system. The most efficient extraction technique was Soxhlet procedure using ethyl acetate for 6 h. Studied samples of O. echioides showed an alkannin/shikonin content in the range of 0.02 -0.24 mg/kg. Other naphthoquinone derivatives (deoxyalkannin/deoxyshikonin and 5,8-dihydroxy-2-(4methyl-6-oxo-5,6-dihydro-2H-pyran-2-yl)-[1,4]naphthoquinone and arnebin-6) were found for the first time in O. echioides and characterized in the extracts using HPLC-MS apparatus equipped with an ESI ionization source.
Optimization of shikonin production in Onosma dichroantha callus using response surface methodology
Plant Cell, Tissue and Organ Culture (PCTOC), 2016
The roots of Onosma dichroantha, contain substantial amounts of shikonin/alkannin derivatives, which are considered as edible natural colors with distinguished anti-inflammatory, antibacterial, antiviral, as well as antitumor activities. In this study, a response surface methodology coupled with numerical optimization modeling techniques were implemented to analyze and optimize the individual as well as interactive effects of two important plant growth regulators (indole acetic acid, IAA and benzylaminopurine, BAP) on the relative growth rate (RGR) and shikonin production of O. dichroantha callus grown on three growth media. Callus growth was best on B5 medium, and shikonin production was best on M9 medium, while White medium was best for synchronized callus growth and shikonin production. Upon statistical optimization, the maximum RGR and shikonin production of callus grown on B5 medium were predicted in 0.93 mg L-1 BAP and 0.16 mg L-1 IAA, on M9 medium in 0.55 mg L-1 BAP and 0.16 mg L-1 IAA, and on White medium in 1.23 mg L-1 BAP and 0.11 mg L-1 IAA. These data can be employed in development of large scale production of shikonin derivatives using O. dichroantha.
Screening of Phytoconstituents in Medicinal Plants and Study of Their Effect in Seed Germination
Amrit Journal, 2023
Due to the existence of bioactive phytochemicals in plants, they have been traditionally utilized as medicine for a very long time. The present study was aimed to determine the phytoconstituents present in ten different plants collected from Katunje-Bhaktapur, Nepal. The methanol extract of the sample was used for analyzing the phytoconstituents present in the plant sample. The result revealed the existence of several phytoconstituents like alkaloid terpenoid, quinine, tannin, saponin and flavonoid in these plants. Similarly, the impact of phytoconstituents on germination of Pisum sativum seeds in water, aqueous extract, methanol, and methanol extract were analyzed. Shoot germination was retarded on all extracts.
Arab Universities Journal of Agricultural Sciences, 2019
This research study was carried out in the plant tissue culture laboratory of the Agricultural Botany Department, Faculty of Agriculture, Ain Shams Univ., Shoubra El-khaima, Cairo, Egypt. Experiments were executed for the duration of two consecutive years 2017 and 2018 on chicory plant. Chicory (Cichorium intybus L.), which belongs to Asteraceae family, is considered as an important medicinal plant due to the presence of many bioactive substances such flavonoids, phenolic compounds, alkaloids, steroids, terpenoids, including (coumarines, cichoriin, esculetin, inulin, sesquiterpene lactones, chicoric acid, caffeic acid and some vitamins). In this research in vitro experiments were carried out using full strength Murashige and Skoog basal medium (MS) supplemented with different combinations of two plant growth regulators; Indole-3-butyric acid (IBA) including two concentrations (0.5-2.0 mg/l) and Naphthalene acetic acid (NAA) comprising four concentrations (0.5-2.0-3.0-5.0 mg/l). An abaxially (lower side) leaf explants (square pieces 0.5 × 0.5 cm) which were taken from 20 days old aseptic chicory seedlings were inoculated to (MS) surface. Initially, chicory seeds were aseptically germinated on half-strength MS medium, after surface sterilization by 70 % (v/v) ethanol for 60 seconds then soaking in 10 % Clorox (0.5% sodium hypochlorite NaOCl) for 10 min to produce the aseptic chicory seedlings which were the source of true leaf explants used in this research study. Total phenolic compounds and flavonoids content were extracted from six-week C. intybus friable callus produced under both light and dark in vitro culture conditions inside a growth chamber incubation room where temperature was adjusted at 25 o C ±1. Total phenolic compounds and flavonoids were determined by spectrophotometric methods. The highest values for their contents were from chicory calli when MS callus induction medium was supplemented with 2 mg/l NAA under total dark condition when compared with the other remaining growth regulator treatment combinations and alternative light regime conditions.
International Journal of Applied Pharmaceutics, 2021
Objective: The research aimed to provide new information regarding the secondary metabolites content of purple and white-purple Orthosiphon aristatus (Blume) Miq. callus, which can then be used as a basis for developing towards cell suspension and ultimately producing secondary metabolites using bioreactors. Methods: Callus induction of two varieties of O. aristatus were performed by inoculating sterile leaf explants grown on Murashige and Skoog basal media supplemented with 2,4-dichlorophenoxyacetis acid 0.4 ppm. The secondary metabolites were analysed and quantified using highperformance liquid chromatography with gradient elution. Results: The results showed the growth of callus two varieties of O. aristatus in growth media MS with 2,4-D 0.4 ppm. Rosmarinic acid content in the acetone extract of the purple variety callus was 1.28% w/w, and the white-purple variety was 2.22% w/w. Conclusion: This study could form the basis for the development of rosmarinic acid production by In vitro culture modification.