Production Of Naphthoquinones Research Articles

Effect of Silica Xerogel Functionalization on Intensification of Rindera graeca Transgenic Roots Proliferation and Boosting Naphthoquinone Production.

Secondary metabolites derived from plants are recognized as valuable products with several successful applications in the pharmaceutical, cosmetic, and food industries. The major limitation to the broader implementation of these compounds is their low manufacturing efficiency. Current efforts to overcome unprofitability depend mainly on biotechnological methods, especially through the application of plant in vitro cultures. This concept allows unprecedented bioengineering opportunities for culture system modifications with in situ product removal. The silica-based xerogels can be used as a novel, porous biomaterial characterized by a large surface area and high affinity to lipophilic secondary metabolites produced by plant tissue. This study aimed to investigate the influence of xerogel-based biomaterials functionalized with methyl, hydroxyl, carboxylic, and amine groups on Rindera graeca transgenic root growth and the production of naphthoquinone derivatives. The application of xerogel-based scaffolds functionalized with the methyl group resulted in more than 1.5 times higher biomass proliferation than for reference untreated culture. The naphthoquinone derivatives' production was noted exclusively in culture systems supplemented with xerogel functionalized with methyl and hydroxyl groups. Applying chemically functionalized xerogels as in situ adsorbents allowed for the enhanced growth and productivity of in vitro cultured R. graeca transgenic roots, facilitating product isolation due to their selective and efficient accumulation.

Selective Impact of MTMS-Based Xerogel Morphology on Boosted Proliferation and Enhanced Naphthoquinone Production in Cultures of Rindera graeca Transgenic Roots.

In situ extraction is a method for separating plant secondary metabolites from in vitro systems of plant biomass cultures. The study aimed to investigate the MTMS-based xerogels morphology effect on the growth kinetics and deoxyshikonin productivity in xerogel-supported in vitro culture systems of Rindera graeca hairy root. Cultures were supplemented with three types of xerogel, i.e., mesoporous gel, microporous gel, and agglomerated precipitate, in the disintegrated or monolithic form. Structure, oil sorption capacity, and SEM analyses for xerogel-based additives were performed. Application of monolithic macroporous xerogel resulted in the highest biomass proliferation, i.e., 5.11-fold fresh biomass increase after four weeks of the screening culture. The highest deoxyshikonin production (i.e., 105.03 µg) was noted when hairy roots were maintained with particles of disintegrated mesoporous xerogel. The detailed kinetics investigations (6-week culture) revealed the highest growth of hairy root biomass and secondary metabolite production, equaling 9.46-fold fresh weight biomass and 204.08 µg deoxyshikonin, respectively. MTMS-based xerogels have been recognized as selective biocompatible scaffolds for boosting the proliferation of transgenic roots or for productivity enhancement of naphthoquinones without detrimental effects on biomass growth, and their successful applicability in in situ removal of secondary plant metabolites has been experimentally confirmed.

Plumbagin, a Plant-derived Naphthoquinone Production in Tissue Cultures of Drosera spatulata Labill

Plumbagin, a Plant-derived Naphthoquinone Production in Tissue Cultures of Drosera spatulata Labill

Production of 3-geranyl-4-hydroxybenzoate acid in yeast, an important intermediate of shikonin biosynthesis pathway.

Shikonin and its derivatives are the main active components in the medicinal plant Arnebia euchroma and possess extensive pharmaceutical properties. In this study, we developed an optimized yeast system to obtain high-level production of 3-geranyl-4-hydroxybenzoate acid (GBA), an important intermediate involved in shikonin biosynthesis pathway. For host selection, recombinant expression of p-hydroxybenzoate:geranyltransferase (PGT) derived from A. euchroma was performed in Saccharomyces cerevisiae WAT11U strain and high yield monoterpene strain. In shake flask culture with 1 mM p-hydroxybenzoate acid (PHBA), they could yield GBA at 0.1567 and 20.8624 mg L-1, respectively. Additionally, AePGT6 showed higher enzymatic activity than its homologs. Moreover, by combining improvement in the homologous mevalonate pathway with reconstruction in the heterologous shikimic pathway, a de novo GBA synthesis pathway was constructed in StHP6tHC with co-overexpressed SctHMG1, optimized EcUbiC and AePGT6. A high titer of 179.29 mg L-1 GBA was achieved in StHP6tHC under shake flask fermentation with 1 mM PHBA. These results suggest that yeast could be engineered systematically to enable an efficient monoterpene-quinone or naphthoquinone production.

Production of naphthoquinones and phenolics by a novel isolate Fusarium solani PSC-R of Palk Bay and their industrial applications

The present study was attempted to enhance the production of naphthoquinones and phenolics by Fusarium solani PSC-R of Palk Bay origin, which exhibited potent antibacterial, antioxidant and dyeing activity. Maximum productivity of naphthoquinones and phenolics was achieved in potato infusion medium supplemented with 2% sucrose. Addition of nitrogen sources to the medium adversely affected the production of both naphthoquinones and phenolics. An initial pH of 5 and incubation at 31°C for six days at 140rpm was found to increase the yield (123.65mg/g of DW), concentration (867.33mg/l) and total naphthoquinones (602.8μM/g DW) by 7.58, 10.44 and 3.68-fold respectively. Similarly, the antioxidant and antibacterial activity associated with the phenolics of PSC-R increased by 1.5-fold in the optimized medium. The obtained results document the effective means of enhanced production of naphthoquinones and phenolics in the suspension culture of F. solani PSC-R at bioreactor level.

Effect of methionine on production of naphthoquinones in Impatiens balsamina root cultures and detection of some secondary metabolites

Context: Lawsone, lawsone methyl ether and 3,3′-methylelnebislawsone are the main active compounds of Impatiens balsamina L. (Balsaminaceae). These compounds possess various pharmacological activities that have been shown to assist with the treatment of skin diseases.Objective: This work focused on increased naphthoquinone production in I. basamina root cultures using methionine feeding.Materials and methods: I. balsamina root cultures were maintained in liquid Gamborg’s B5 medium supplemented with 0.1 mg/L α-naphthalene acetic acid, 0.1 mg/L kinetin, 1.0 mg/L 6-benzyladenine and 20 g/L sucrose. The effect of methionine concentration (50, 100, 300, 500 and 1000 mg/L) on naphthoquinone production of I. basamina root cultures was determined. Isolation of secondary metabolites from I. balsamina root cultures was also carried out.Results and discussion: Feeding of 300 mg/L methionine to the root cultures at the beginning of the growth cycle increased the production of 3,3′-methylelnebislawsone almost two-fold (0.63 mg/g dry weight, compared to the control group 0.32 mg/g dry weight). Optimization of the feeding conditions showed that adding 500 mg/L methionine to a 21-day old root cultures increased production of lawsone methyl ether and 3,3′-methylenebislawsone up to 2.6- and 3.1-fold higher, respectively, compared to the controls. In addition, various pharmacologically interesting secondary metabolites were isolated from I. balsamina root cultures, such as a flavonoid, luteolin, a naphthoquinone, 2,3-dihydroxy-1,4-naphthoquinone, and a triterpenoid, echinocystic acid. This is the first report of the occurrence of these compounds in this plant.

Cynoglossum columnae Ten. – chemical profiling through biotechnological and phytochemical approach

Cynoglossum columnae Ten. (Boraginaceae) is an annual species of Mediterranean region. Plants of this family are investigated for naphthoquinone red pigments, found in the underground parts, which are bioactive constituents known as: wound healing, anti-inflammatory, antimicrobial, antitumor agents [1]. Boraginaceae family is also known for its content of hepatotoxic pyrrolizidine alkaloids (PAs) [2]. Three groups of in vitro root cultures were obtained and investigated: the natural roots cut off from seedlings, natural roots regenerated from shoots in the following passages and hairy roots obtained as a result of transformation with Agrobacterium rhizogenes– ATCC 15834 strain. The roots were cultured in various liquid media (ex.: MS, B5, LS, DCR) and tested for their growth and production of naphthoquinones. The best results gave DCR medium [3] with twice reduced amount of all components and full sugar value. Preliminary phytochemical analysis was performed using RP-HPLC DAD method and showed the presence of six naphtoquinone derivatives in natural roots and their post culture media (DCR/2); none in the transformed root cultures was observed. Phytochemical investigation in plants cultivated in nature was made and six PAs have been isolated and determined by modern spectroscopic methods as rinderine, 3′-acetylrinderine and echinatine in the form of both their N-oxides as well as their bases after reduction. Sixteen more, known PAs, were also identified after reduction as echimumiline, heliosupine, heliotridine, echinatine, rinderine, retronecine, integerrimine and triangularicine type derivatives.

Stimulation of naphthoquinone production in Impatiens balsamina root cultures by methyl jasmonate

Lawsone (1), lawsone methyl ether (2) and methylene-3,3′-bilawsone (3) are pharmacological active naphthoquinones (NQs) found in Impatiens balsamina L. (Balsaminaceae) [1,2]. I. balsamina root cultures have been established from the young leaf explants in liquid B5 medium supplied with 0.1mg/l α-naphthalene acetic acid, 1.0mg/l Kinetin, 2.0mg/l and 6-benzyladenine [3]. However, these root cultures produced very low levels of (2). The main aims of this study were therefore to increase NQ production in I. balsamina root cultures using methyl jasmonate (MJ) as an elicitor.

Increased production of naphthoquinones in Impatiens balsamina root cultures by elicitation with methyl jasmonate

Impatiens balsamina root cultures were treated with yeast extract (YE), Candida albicans homogenate (CAH), Trichophyton rubrum homogenate (TRH), chitosan (CHI) and methyl jasmonate (MJ). Different elicitors, depending on concentrations used exerted differential effects on the production of the three main naphthoquinones, lawsone (2-hydroxy-1,4-naphthoquinone), lawsone methyl ether and methylene-3,3'-bilawsone. Treatment with MJ (400microM) was capable of increasing production of lawsone, and lawsone methyl ether up to 8.6- and 11.3-fold higher, respectively, compared to the level in untreated cultures. Treatment of 21-day-old root cultures with 300microM MJ for 36h resulted in the production of 10.0, 0.78 and 0.23mg/g DW of lawsone, its methyl ether and methylene-3,3'-bilawsone, respectively. Such levels are sufficient for commercial production.

Effects of donor plants and plant growth regulators on naphthoquinone production in root cultures of Impatiens balsamina

The effects of type of explant (leaves and roots), donor plants, and plant growth regulators on naphthoquinone (NQ) production of Impatiens balsamina L. root cultures were evaluated. The root cultures were initiated in liquid Gamborg’s B5 medium supplemented with 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 kinetin (Kn) and 1.0 mg l−1 6-benzyladenine (BA). The present investigation indicated that the root cultures established from the leaf explants produced higher total NQ content [1.01 ± 0.046 mg/g dry weight (DW)] than those established from the root explants (0.62 ± 0.023 mg/g DW). The leaf explants of four I. balsamina strains including white flower plant (IbW), pink flower plant (IbP), violet flower plant (IbV) and red flower plant (IbR) were used to establish the root cultures. Based on HPLC analysis, IbP strain produced the highest total NQ content (3.39 ± 0.072 mg/g DW), while IbR strain produced the lowest one (1.45 ± 0.055 mg/g DW). The root cultures established from the IbP explant were capable of producing higher content of total NQs (2.76 ± 0.093 mg/g DW) than those established from the other strains. The results suggest that the tissue cultures initiated from the high-yielding donor plants should be capable of producing higher content of secondary compounds than those initiated from low-yielding donor plants. In addition, plant growth regulator manipulation exhibited that a combination of 0.1 mg l−1 NAA, 1.0 mg l−1 Kn and 2.0 mg l−1 BA is capable of increasing NQ production (2.97 ± 0.072 mg/g DW) in I. balsamina root cultures.

Effects of Different Carbon and Nitrogen Sources on Naphthoquinone Production of Cordyceps unilateralis BCC 1869

The production of six naphthoquinone derivatives, erythrostominone, deoxyerythrostominone, 4-O-methyl erythrostominone, epierythrostominol, deoxyerythrostominol, and 3,5,8-trihydroxy-6-methoxy-2-(5-oxohexa- 1,3-dienyl)-1,4-naphthoquinone, was examined during the growth of Cordyceps unilateralis BCC 1869 on different carbon and nitrogen sources. Erythrostominone production by the fungus accounted for more than 50% of total naphthoquinones, but production of each of the other five derivatives accounted for less than 20% of total naphthoquinones. The highest volumetric production rate of erythrostominone and overall naphthoquinone production rate were obtained on mannose as a sole carbon source and ammonium sulfate as a sole nitrogen source (4922.4 +/- 118.8 mg/[L.d] and 5.03 g/[L.d], respectively). The highest growth rate was obtained on arabinose (0.043 h-1), whereas the maximum overall naphthoquinone concentration was obtained on lactose (2 g/L) at 237 h. These naphthoquinones were produced with no pH control and were first detected at a pH of about 3.0 to 4.0. These results suggest that carbon and nitrogen influenced directly the production of naphthoquinones.

A study of the factors influencing non-genetic variation in a strain of Fusarium oxysporum.

SUMMARY: The colour variation of a strain of Fusarium oxysporum Schlecht ex Fr. was found to depend on the production of carotenoids and naphthoquinones. The carotenoids were only produced as a result of exposure to light, whereas the production of naphthoquinones, which were pH indicators, was mainly regulated by the carbon/nitrogen ratio of the medium. The morphogenetic effects of radiation and of carbon/nitrogen ratio were also considered. A low carbon/nitrogen ratio favoured chlamydospore formation and aerial mycelium, whereas ultraviolet radiation, and to a lesser extent light, promoted macrospore and sclerotium production.