Profiling the chemical content of Opuntia ficus-indica flowers by HPLC–PDA-ESI-MS and GC/EIMS analyses

The qualitative and quantitative analysis of flavonoids from Opuntia ficus-indica flowers methanol extract from the Mediterranean area is described. On the basis of HPLC–PDA-ESI-MS/MS analysis seven compounds have been identified as kaempferol, quercetin, and isorhamnetin glycosylated derivatives. The total amount of flavonoids of O. ficus-indica flowers was 81.75 mg/1 g of fresh plant material, with isorhamnetin 3-O-robinobioside being the major component (52.22%). The plant flowers volatiles composition was also characterized and a total of 18 components were identified. The main constituents were found to be germacrene D (12.6%), 1-hexanol (12.3%), n-tetradecane (9.1%) and decanal (8.2%).     

Enhanced extraction genistein from pigeon pea [Cajanus cajan (L.) Millsp.] roots with the biotransformation of immobilized edible Aspergillus oryzae and Monacus anka and antioxidant activity evaluation

A new method of enhanced extraction genistein from pigeon pea [Cajanus cajan (L.) Millsp.] roots with the biotransformation of immobilized edible Aspergillus oryzae and Monacus anka, was investigated. It showed that immobilized Aspergillus oryzae and Monacus anka on sodium alginate effectively supported the highest genistein extraction yield by screening microorganism tests. After biotransformation process with immobilized Aspergillus oryzae and Monacus anka under 30 °C, pH 6.0, 2 days, liquid-solid ratio 12: 1 (mL/g), the extraction yield of genistein reached 1.877 mg/g, which was 2.65-fold to that of normal extraction yield. Moreover, IC₅₀ values of the extracts measured by DPPH-radical scavenging test and β-Carotene-linoleic acid bleaching test were 0.737 mg/mL and 0.173 mg/mL (control sample 1.117 mg/mL and 0.216 mg/mL), respectively. SOD (Super Oxygen Dehydrogenises) activity of the extracts treated with immobilized microorganism which was stronger than that of the untreated pigon pea roots (1.44 U/mg) at the concentration of protein (0.9375 μg/mL) was 1.83 U/mg. The developed method could be an alternative method for the enhanced extraction of genistein from plants and could be potentially applied in the food industry     

Kaempferol from the leaves of Apocynum venetum possesses anxiolytic activities in the elevated plus maze test in mice

The present work evaluated the anxiolytic activity of an aqueous extract of Apocynum venetum L. (Apocynaceae) and bioguided its fractionation using the elevated plus maze (EPM) in mice as a model of anxiety. A single treatment of AV extract markedly increased the percentage time spent on the open arms of the EPM in two distinct concentration ranges of 22.5–30 and 100–125 mg/kg p.o., respectively, indicating a putative anxiolytic-like activity. Fractions showing anxiolytic effects in concentrations equal to 30 or 125 mg/kg of whole extract were antagonized using the benzodiazepine antagonist flumazenil (3 mg/kg i.p.) or the 5-HT_1A receptor antagonist WAY-100635 (0.5 mg/kg i.p.). All active fractions in a concentration equal to 125 mg/kg were effectively blocked by the benzodiazepine antagonist flumazenil, while the anxiolytic activities of fractions in the lower dose equivalent to 30 mg/kg of whole extract were inhibited by the 5-HT_1A receptor antagonist WAY-100635. Through further separation of AV fractions it was possible to isolate and characterize the flavonol kaempferol which showed an anxiolytic-like activity in concentrations from 0.02 to 1.0 mg/kg p.o. The anxiolytic activity of kaempferol was partially antagonized by concomitant administration of flumazenil, but not by WAY-100635. In conclusion, our study clearly demonstrates that AV extract possesses anxiolytic-like activity and that at least one of its flavonoids, kaempferol, can elicit the same kind of neuropharmacological activity.     

Temporal lag between gene expression and metabolite accumulation in flavonol biosynthesis of Arabidopsis roots

The temporal lag between gene expression and metabolite accumulation has been estimated in flavonol biosynthesis, but the time difference between these events is unclear. In the present study, we investigated the expression of flavonol biosynthetic genes ELONGATED HYPOCOTYL5, MYELOBLASTOSIS PROYEIN12/PRODUCTION OF FLAVONOL GLYCOSYDES1, CHALCONE SYNTHASE, CHALCONE ISOMERASE, FLAVANONE 3-HYDROXYLASE, and FLAVONOL SYNTHASE1, and the accumulation of flavonol glycosides (kaempferol and quercetin glycosides) in time-series samples of Arabidopsis thaliana roots. All genes started to be expressed within 3 h after sequential light irradiation (HAS) and reached their maximum expression levels at 12 HAS, and the accumulation of the flavonol glycosides started at 6 HAS. Metabolome analysis using liquid chromatography-mass spectrometry showed that the accumulation of kaempferol 3-O-glucoside-7-O-rhamnoside and kaempferol 3-O-rhamnosyl (1 → 2) glucoside-7-O-rhamnoside reached their maximum levels at 48 HAS, whereas other flavonol glycosides, such as kaempferol/quercetin 3-O-rhamnoside-7-O-rhamnoside, quercetin 3-O-glucoside-7-O-rhamnoside and quercetin 3-O-rhamnosyl (1 → 2) glucoside-7-O-rhamnoside, increased gradually until 96 HAS. These results show that the expression of the flavonol genes is an early response against light exposure, and that the accumulation of the flavonol glycosides is a late response.     

Metabolic engineering of Saccharomyces cerevisiae to improve 1-hexadecanol production

Fatty alcohols are important components of a vast array of surfactants, lubricants, detergents, pharmaceuticals and cosmetics. We have engineered Saccharomyces cerevisiae to produce 1-hexadecanol by expressing a fatty acyl-CoA reductase (FAR) from barn owl (Tyto alba). In order to improve fatty alcohol production, we have manipulated both the structural genes and the regulatory genes in yeast lipid metabolism. The acetyl-CoA carboxylase gene (ACC1) was over-expressed, which improved 1-hexadecanol production by 56% (from 45 mg/L to 71 mg/L). Knocking out the negative regulator of the INO1 gene in phospholipid metabolism, RPD3, further enhanced 1-hexadecanol production by 98% (from 71 mg/L to 140 mg/L). The cytosolic acetyl-CoA supply was next engineered by expressing a heterologous ATP-dependent citrate lyase, which increased the production of 1-hexadecanol by an additional 136% (from 140 mg/L to 330 mg/L). Through fed-batch fermentation using resting cells, over 1.1 g/L 1-hexadecanol can be produced in glucose minimal medium, which represents the highest titer reported in yeast to date.     

Optimisation of preparative HPLC separation of four isomeric kaempferol diglycosides from Prunus spinosa L. by application of the response surface methodology

A fast and efficient preparative HPLC-PDA method was developed for the separation and isolation of four rare isomeric kaempferol diglycosides from leaves of Prunus spinosa L. The separation procedure of the enriched diglycoside fraction of the 70% (v/v) aqueous methanolic leaf extract was first optimised on analytical XBridge C18 column (100 mm × 4.6 mm i.d., 5 μm) and central composite design combined with response surface methodology was utilized to establish the optimal separation conditions. The developed method was directly transferred to preparative XBridge Prep C18 column (100 mm × 19 mm i.d., 5 μm) and the final separation was accomplished by isocratic elution with 0.5% acetic acid-methanol-tetrahydrofuran (75.2:16.6:8.2, v/v/v) as the mobile phase, at a flow rate of 13.6 mL/min, in less than 12 min for a single run. Under these conditions, four flavonoid diglycosides: kaempferol 3-O-α-l-arabinofuranoside-7-O-α-l-rhamnopyranoside, kaempferol 3,7-di-O-α-l-rhamnopyranoside (kaempferitrin), and reported for the first time for P. spinosa kaempferol 3-O-β-d-xylopyranoside-7-O-α-l-rhamnopyranoside (lepidoside) and kaempferol 3-O-α-l-arabinopyranoside-7-O-α-l-rhamnopyranoside, were isolated in high separation yield (84.8–94.5%) and purity (92.45–99.79%). Their structures were confirmed by extensive 1D and 2D NMR studies. Additionally, the UHPLC-PDA-ESI–MS³ qualitative profiling led to the identification of twenty-one phenolic compounds and confirmed that the isolates were the major components of the leaf material.     

Fast biodegradation of long chain n-alkanes and crude oil at high concentrations with Rhodococcus sp. Moj-3449

Biodegradation of long chain n-alkanes and crude oil with fast rate and high concentration are desirable for bioremediation, especially in heavily oil-polluted areas, and enhanced oil recovery. We discovered Rhodococcus sp. Moj-3449 with such unique abilities by screening microorganisms for the growth on n-hexadecane at 30 mg/mL. The new strain grew very fast on 120 mg/mL of n-hexadecane giving a cell density of 14.7 g cdw/L after only 2 days’ incubation. During the growth with this strain, the oil–water phases were rapidly emulsified, giving rise to tolerance to high alkane concentration (250 mg/mL) and fast growth rate of 0.10–0.20 h^?1 for alkane concentration of 1–180 mg/mL. The degraded concentration of n-hexadecane increased linearly with the initial alkane concentration (1–250 mg/mL). Incubation on n-hexadecane at 250 mg/mL for 7 days gave a cell density of 13.5 g cdw/L and degraded 124 mg/mL of n-hexadecane. The strain grew also fast on n-dodecane (C12), n-tetradecane (C14), and n-octadecane (C18), with degradation preference of C14 (=C16) > C12 > C18. Different from many alkane-degrading strains, Rhodococcus sp. Moj-3449 was found to have subterminal oxidation pathway. Rhodococcus sp. Moj-3449 degraded also crude oil fast at 60–250 mg/mL, with a wide range of n-alkanes (C10–C35) as substrates in which C14–C19 are preferred. The degradation ability increased with initial oil concentration from 60 to 150 mg/mL and slightly decreased afterwards. Incubation on 150 mg/mL of crude oil for 7 days degraded 37% of n-alkanes. The outstanding ability of rapidly degrading long chain n-alkanes and crude oil at high concentration makes Rhodococcus sp. Moj-3449 potentially useful for bioremediation and microbial enhanced oil recovery.     

Metabolic engineering of Saccharomyces cerevisiae for production of ginsenosides

Ginsenosides are the primary bioactive components of ginseng, which is a popular medicinal herb and exhibits diverse pharmacological activities. Protopanaxadiol is the aglycon of several dammarane-type ginsenosides, which also has anticancer activity. For microbial production of protopanaxadiol, dammarenediol-II synthase and protopanaxadiol synthase genes of Panax ginseng, together with a NADPH-cytochrome P450 reductase gene of Arabidopsis thaliana, were introduced into Saccharomyces cerevisiae, resulting in production of 0.05 mg/g DCW protopanaxadiol. Increasing squalene and 2,3-oxidosqualene supplies through overexpressing truncated 3-hydroxyl-3-methylglutaryl-CoA reductase, farnesyl diphosphate synthase, squalene synthase and 2,3-oxidosqualene synthase genes, together with increasing protopanaxadiol synthase activity through codon optimization, led to 262-fold increase of protopanaxadiol production. Finally, using two-phase extractive fermentation resulted in production of 8.40 mg/g DCW protopanaxadiol (1189 mg/L), together with 10.94 mg/g DCW dammarenediol-II (1548 mg/L). The yeast strains engineered in this work can serve as the basis for creating an alternative way for production of ginsenosides in place of extraction from plant sources.     

Acylated flavonoid tetraglycoside from Planchonia careya leaves

Phytochemical investigations of the aqueous extract of Planchonia careya leaves revealed two known flavonol glycosides, kaempferol 3-O-gentiobioside (1) and quercetin 3-O-glucoside (isoquercitrin) (2), and a novel acylated kaempferol tetraglycoside, kaempferol 3-O-[α-rhamnopyranosyl(1 → 3)-(2-O-p-coumaroyl)]-β-glucopyranoside, 7-O-[α-rhamnopyranosyl-(1 → 3)-(4-O-p-coumaroyl)]-α-rhamnopyranoside (3). Structural elucidation was achieved using UV, NMR, and mass spectrometry.     

Variation in polyphenolic profile and in vitro antioxidant activity of eastern teaberry (Gaultheria procumbens L.) leaves following foliar development

Seasonal dynamics in the polyphenolic composition, antioxidant activity, and their relationships during plant development were evaluated for eastern teaberry (Gaultheria procumbens L.) leaves, a traditional herbal medicine of North American natives. With the complementary UHPLC-PDA-ESI-MS³, HPLC-PDA-fingerprint, Folin-Ciocalteau, and n-butanol/HCl assays of methanol-water (75:25, v/v) extracts, the dried leaf samples harvested monthly across the growing season under Polish climate conditions were found rich in structurally diverse polyphenols (149.2–210.7 mg/g DW) including the dominating salicylates (64.6–107.5 mg/g DW), proanthocyanidins (53.0–66.8 mg/g DW), and flavonoids (17.3–25.3 mg/g DW), and the accompanying chlorogenic acid isomers (2.4–4.4 mg/g DW) and simple phenolic acids (0.9–1.1 mg/g DW). Among 28 detected analytes, gaultherin (64.6–107.5 mg/g DW), miquelianin (14.6–21.1 mg/g DW), procyanidin A-type trimer (5.5–9.5 mg/g DW), and (–)-epicatechin (5.8–7.8 mg/g DW) were the most abundant. The phenolic levels and antioxidant activity parameters in the DPPH (EC₅₀, 15.0–18.2 μg DW/mL; 0.95–1.16 mmol Trolox equivalents/g DW) and FRAP (2.3–3.4 mmol Fe ²⁺/g DW; 0.86–1.26 mmol Trolox equivalents/g DW) assays showed parallel seasonal trends with maxima in September and October. As the subsequent correlation studies confirmed the determinative impact of polyphenols on the leaf antioxidant activity and its seasonal fluctuations, the Fall season could be recommended as optimal for harvesting the plant material for medicinal purposes and cost-effective production of natural health products.     

Molecular characterization and molasses fermentation performance of a wild yeast strain operating in an extremely wide temperature range

Molasses fermentation performance by both a cryotolerant and a thermophilic yeast (strain AXAZ-1) isolated from grapes in Greece was evaluated in an extremely wide temperature range (3–40 °C). Sequence analysis of the 5.8S internal transcribed spacer and the D1/D2 ribosomal DNA (rDNA) regions assigned isolate to Saccharomyces cerevisiae. Restriction fragment length polymorphism of the mitochondrial DNA showed that strain AXAZ-1 is genetically divergent compared to other wild strains of Greek origin or commercial yeast starters. Yeast cells growing planktonically were capable of fermentation in a wide temperature spectrum, ranging from 3 °C to 38 °C. Immobilization of yeast on brewer’s spent grains (BSG) improved the thermo-tolerance of the strain and enabled fermentation at 40 °C. Time to complete fermentation with the immobilized yeast ranged from 20 days at 3 to 38 h at 40 °C. The daily ethanol productivity reached maximum (58.1 g/L) and minimum (2.5 g/L) levels at 30 and 3 °C, respectively. The aroma-related compounds’ profiles of immobilized cells at different fermentation temperatures were evaluated by using solid phase microextraction (SPME) gas chromatography–mass spectrometry (GC–MS). Molasses fermentation resulted in a high quality fermentation product due to the low concentrations of higher and amyl alcohols at all temperatures tested. Strain AXAZ-1 is very promising for the production of ethanol from low cost raw materials, as it was capable to perform fermentations of high ethanol concentration and productivities in both low and high temperatures.     

Aerobic degradation of 2-picolinic acid by a nitrobenzene-assimilating strain: Streptomyces sp. Z2

Streptomyces sp. Z2 was isolated from nitrobenzene contaminated activated sludge, which utilized nitrobenzene as a sole source of carbon, nitrogen, and energy under aerobic condition. It was found that besides nitrobenzene strain Z2 can degrade 2-picolinic acid. Strain Z2 completely degraded 2-picolinic acid with initial concentration of 500 mg/L, 1000 mg/L, 1500 mg/L, 2000 mg/L, 2500 mg/L, and 3000 mg/L within 36 h, 50 h, 72 h, 100 h, 136 h, and 180 h, respectively. Kinetics of 2-picolinic acid degradation was described using the Andrews equation. The kinetic parameters were as follows: q_max = 3.81 h^?1, K_s = 83.10 mg/L, and K_i = 252.11 mg/L. During the biodegradation process, Z2 transformed 2-picolinic acid into a product which was identified as 6-hydroxy picolinic acid by UV–vis spectrometry, ¹H nuclear magnetic resonance spectroscopy, and mass spectrometry. 6-Hydroxy picolinic acid was then cleaved and mineralized with release of ammonia.     

Optimization of the medium for glutathione production in Saccharomyces cerevisiae

As a powerful statistical experimental design, uniform design (UD) method has been successfully applied in various fields such as fermentation industry, pharmaceuticals, and others. In this paper, UD was applied to optimize the medium composition for glutathione production in shake-flask culture of Saccharomyces cerevisiae T65. The experiments of nine factors (glucose, yeast extract, peptone, malt extract, molasses, MgSO₄, ZnSO₄, (NH₄)₂HPO₄ and thiamine) and nine levels were carried out according to the uniform design table U₂₇(9⁹). The experimental data was analyzed to obtain the regression model and the optimal medium composition was achieved by optimization with UD 3.0 software. The optimal medium consisted of 70 g/L glucose, 3 g/L yeast extract, 5 g/L peptone, 70 g/L malt extract, 20 g/L molasses, 5.6 g/L MgSO₄, 16 mg/L ZnSO₄, 7 g/L (NH₄)₂HPO₄ and 0.2 mg/L thiamine. The GSH yield at the optimal point achieved 74.6 mg/L, which was 1.81 times higher than that of the control. The application of UD method resulted in enhancement in GSH production.     

Variation of quercetin glycoside derivatives in three onion (Allium cepa L.) varieties

The aim of this study was to quantify the contents of individual quercetin glycosides in red, yellow and chartreuse onion by High Performance Liquid Chromatography (HPLC) analysis. Acid hydrolysis of individual quercetin glycosides using 6 M hydrochloric acid guided to identify and separate quercetin 7,4′-diglucoside, quercetin 3-glucoside, quercetin 4′-glucoside, and quercetin. The contents of total quercetin glycosides varied extensively among three varieties (ranged from 16.10 to 103.93 mg/g DW). Quercetin was the predominant compound that accounted mean 32.21 mg/g DW in red onion (43.6% of the total) and 127.92 mg/g DW in chartreuse onion (78.3% of the total) followed by quercetin 3-glucoside (28.83 and 24.16 mg/g DW) respectively. Quercetin 3-glucoside levels were much higher in yellow onion (43.85 mg/g DW) followed by quercetin 30.08 mg/g DW. Quercetin 4′-glucoside documented the lowest amount that documented mean 2.4% of the total glycosides. The varied contents of glycosides present in the different onion varieties were significant.     

Prepubertal octylphenol exposure up-regulate BRCA1 expression,down-regulate ERα expression and reduce rat mammary tumorigenesis

Background: Endogenous estrogens play an important role in the development of breast cancer. Octylphenol (OP) and genistein (GEN) are estrogen-like chemicals. Prepubertal estradiol and genistein exposure can up-regulate BRCA1 mRNA in mammary gland and reduce futuer breast cancer risk. In the present study, the effects of prepubertal exposure to high-dose OP and GEN on mammary carcinogenesis and the association with the expression of BRCA1 and ERα were investigated. Methods: Prepubertal female Sprague–Dawley rats were exposed to 20, 40, 80 mg/kg OP daily from postnatal day (PND) 22–28, subsequently, the rats were given a single dose of 100 mg/kg 7,12-dimethylbenz [a] anthracene (DMBA) on PND42 to induce mammary tumor. Results: The incidence of DMBA-induced mammary tumors significantly decreased when rats were treated with 40 mg/kg OP. BRCA1 mRNA and protein expression were found up-regulated and ERα expression was down-regulated in the mammary tumor when rats were exposed to 40 mg/kg octylphenol. Conclusion: Exposure 40 mg/kg octylphenol can reduce later breast cancer risk in prepubertal Sprague–Dawley rats, the protective effect of OP is associated with persistent up-regulation of BRCA1 and down-regulation of ERα in the mammary tumor.     

Optimization of a biotransformation process to produce 4-(2,3,5,6-tetramethylpyrazine-1)-4′-demethylepipodophyllotoxin

The developed tandem biotransformation process for the directional biosynthesis of a designed compound 4-(2,3,5,6-tetramethylpyrazine-1)-4′-demethylepipodophyllotoxin (4-TMP-DMEP) by Alternaria alternata S-f6 was systematically optimized. 28 °C of culture temperature and 120 rpm of rotary shaker speed were suitable for the accumulation of 4-TMP-DMEP. The production (i.e., 11.1 ± 1.4 mg/L) of 4-TMP-DMEP was remarkably improved by using an initial yeast extract concentration of 2.5 g/L. 2.0 g/L of Span 80 was beneficial for the 4-TMP-DMEP production (i.e., 25.0 ± 1.5 mg/L). Furthermore, the 4-TMP-DMEP production was remarkably improved by one pulse feeding of 50 mg/L of DMEP on day 6 and two pulse feedings of 40 mg/L of TMP on days 8 and 14 when its residual level was below 50 mg/L and 10 mg/L, respectively. The 4-TMP-DMEP production of 45.1 ± 1.6 mg/L was obtained in the fed-batch biotransformation process, which was enhanced by 726% and 256%, comparing to that (i.e., 5.4 ± 0.4 mg/L and 0.9 mg/L/day) obtained in the batch biotransformation before optimization.     

High performance liquid chromatographic determination of plasma free and total tazobactam and piperacillin

A high-pressure liquid chromatography (HPLC) method with ultraviolet detection was developed for the measurement of plasma free and total tazobactam and piperacillin. This method is simple and fast, requiring only 11 min for the HPLC run and a sample preparation of about 11 min for total drugs and 10 min for free drugs. The procedure for the assay involves the treatment of plasma with acetonitrile for total drugs determination, and the use of a centrifugal filter device to deproteinize plasma for free drugs determination. The HPLC column, a Hypersil-ODS, was equilibrated with an eluent mixture composed of acetonitrile–potassium phosphate (pH 2.6). CVs for repeatability of tazobactam and piperacillin measurements ranged from 4.30 to 6.60; CVs for reproducibility ranged from 5.60 to 9.40. Mean analytical recoveries ranged from 100.4 to 103%. A linear relationship was obtained between peak area and drugs concentration in the range studied (0–62.5 mg/L for tazobactam and 0–500 mg/L for piperacillin). The equation for regression line were y = 19x ? 1.4 for tazobactam and y = 1.7x ? 0.9 for piperacillin; correlation coefficients were >0.999. The lower limit of quantitation (LLQ) for standard samples was about 0.12 mg/L for tazobactam and 0.49 mg/L for piperacillin, respectively. The lower limit of detection (LLD) was 0.06 mg/L for tazobactam and 0.24 mg/L for piperacillin. This HPLC assay for tazobactam and piperacillin is sensitive and accurate, and provides a reliable determination of both free and total tazobactam and piperacillin in human plasma, thus allowing the determination of these analytes in patients receiving tazocillin therapy.     

Changes of ginsenosides in Korean red ginseng (Panax ginseng) fermented by Lactobacillus plantarum M1

To obtain microorganisms for the microbial conversion of ginsenosides in red ginseng powder (RGP), Lactobacillus species (M1–M4 and P1–P4) were isolated from commercial ginseng products. Strain M1 was determined to be L. plantarum by 16S rRNA sequencing. Red ginseng powder (RGP) fermented by L. plantarum M1 had a high total content of ginsenosides (142.4 mg/g) as compared to the control (121.8 mg/g). In particular, the ginsenoside metabolites Rg3, Rg5, Rk1, Compound K (CK), Rh1, and Rg2 showed a high level in the fermented RGP (65.5 mg/g) compared to the control (32.7 mg/g). During fermentation for 7 days, total sugar content decreased from 8.55 mg/g to 4 mg/g, uronic acid content reached its maximum (53.43 μg/g) at 3 days, and total ginsenoside content increased to 176.8 mg/g at 4 days. In addition, ginsenoside metabolites increased from 38.0 mg/g to 83.4 mg/g at 4 days of fermentation. Using everted instestinal sacs of rats, the fermented red ginseng showed a high transport level (10.3 mg of polyphenols/g sac) compared to non-fermented red ginseng (6.67 mg of polyphenols/g sac) after 1 h. These results confirm that fermentation with L. plantarum M1 is very useful for preparing minor ginsenoside metabolites while being safe for foods.     

Galanthamine production by Leucojum aestivum in vitro systems

The callus induction from young fruits of Leucojum aestivum was performed on Murashige–Skoog nutrient medium supplemented with 4 mg/L 2,4 dichlorphenoxyacetic and 2 mg/L 6-benzylamynopurine. Further, by planting the obtained calluses on the same nutrient medium supplemented with 1.15 mg/L α-naphtylacetic acid and 2.0 mg/L 6-benzylamynopurine shoot cultures were established. The growth and galanthamine accumulation of obtained L. aestivum in vitro systems were studied. It was established that the amount of accumulated galanthamine strongly depended on the level of the differentiation. The maximum yield of biomass (17.8 g/L) and the maximum amount of accumulated galanthamine (2.5 mg/L) were achieved after day 35 of submerged cultivation of L. aestivum 80 shoot culture, performed under illumination. Data concerning the time courses of the utilization of the main nutrient components of the medium during cultivation of L. aestivum shoot culture are presented as well.