Antioxidant activity and phenolic profile in filamentous cyanobacteria: the impact of nitrogen

In the present study, ethanolic extracts of ten cyanobacterial strains cultivated under different nitrogen conditions were assessed for the phenolic content and antioxidant activity. The amount of detected phenolic compounds ranged from 14.86 to 701.69 μg g^?1 dry weight (dw) and HPLC-MS/MS analysis revealed gallic acid, chlorogenic acid, quinic acid, catechin, epicatechin, kaempferol, rutin and apiin. Only catechin, among the detected phenolics, was present in all the tested strains, while quinic acid was the most dominant compound in all the tested Nostoc strains. The results also indicated the possibility of increasing the phenolic content in cyanobacterial biomass by manipulating nitrogen conditions, such as in the case of quinic acid in Nostoc 2S7B from 70.83 to 594.43 μg g^?1 dw. The highest radical scavenging activity in DPPH assay expressed Nostoc LC1B with IC₅₀ value of 0.04?±?0.01 mg mL^?1, while Nostoc 2S3B with IC₅₀ =?9.47?±?3.61 mg mL^?1 was the least potent. Furthermore, the reducing power determined by FRAP assay ranged from 8.36?±?0.08 to 21.01?±?1.66 mg AAE g^?1, and it was significantly different among the tested genera. The Arthrospira strains exhibited the highest activity, which in the case of Arthrospira S1 was approximately twofold higher in comparison to those in nitrogen-fixing strains. In addition to this, statistical analysis has indicated that detected phenolics were not major contributor to antioxidant capacities of tested cyanobacteria. However, this study highlights cyanobacteria of the genera Nostoc, Anabaena, and Arthrospira as producers of antioxidants and phenolics with pharmacological and health-beneficial effects, i.e., quinic acid and catechin in particular.     

Isolation of Natural Naphthoquinones from <Emphasis Type="Italic">Juglans regia</Emphasis> and In Vitro Antioxidant and Cytotoxic Studies of Naphthoquinones and the Synthetic Naphthofuran Derivatives

The naphthoquinones and their derivatives containing hydroxyl group exhibit wide range of pharmacological activities, such as antioxidant, antibacterial, antiviral, anticancer, antimalarial, and antifungal activities. In particular, the antioxidant and anticancer behaviors of these compounds continue to draw attention of researchers. In the present communication, three natural naphthoquinones—juglone, lawsone, and plumbagin—isolated from the chloroform extract of nutshells of Juglans regia Linn. and two 1,4-naphthoquinone derivatives—ethyl-5-hydroxynaphtho[ 1,2-b]furan-3-carboxylate and diethylnaphtho[1,2-b:4,3-b′]difuran-3,4-dicarboxylate—and three 5-hydroxy- 1,4-naphthoquinone derivatives—diethyl-7-hydroxynaphtho[1,2-b:4,3-b']difuran-3,4-dicarboxylate,4-ethoxycarbonyl- 7-hydroxynaphtho[1,2-b:4,3-b']difuran-3-carboxylic acid, and 7-hydroxynaphtho[1,2-b:4,3-b']difuran-3,4- dicarboxylic acid were synthesized and examined for their in vitro antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) bioassays. In addition, the cytotoxicity test using human hepatocellular liver carcinoma cell line (HepG2) was carried out for all the compounds. The 5-hydroxy-1,4-naphthoquinone derivatives displayed almost equivalent scavenging activity in DPPH assay and higher activity in ABTS assay relative to ascorbic acid. On the other hand, naphthoquinones Juglone and Plumbagin showed lesser antioxidant activity, but higher cytotoxic activity than naphthofurans except for diethyl naphtho[1,2-b:4,3-b′]difuran-3,4-dicarboxylate, which showed excellent cytotoxic activity.     

Improvement of shoot proliferation and comparison of secondary metabolites in shoot and callus cultures of <Emphasis Type="Italic">Phlomis armeniaca</Emphasis> by LC-ESI-MS/MS analysis

Phlomis armeniaca Willd. is a medicinal plant in the Lamiaceae family endemic to Turkey. The present study describes efficient plant regeneration and callus induction protocols for P. armeniaca and compares phenolic profiles, total phenol and flavonoid contents, and free radical scavenging activity of in vitro-derived tissues. Stem node explants from germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with 75 plant growth regulator (PGR) combinations. The highest shoot number per explant, frequency of shoot proliferation, and frequency of highly proliferated, green, compact callus were obtained on MS medium containing 0.25 mg L^?1 thidiazuron (TDZ) and 0.25 mg L^?1 indole-3-acetic acid (IAA). The best root formation was on MS basal medium (control). Methanol extract of leaves obtained from regenerants contained higher total phenol and flavonoid contents than the callus extract. The callus extract showed stronger free radical scavenging activity than leaves with IC₅₀ [concentration inhibiting 50% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical] values of 4.30 ± 0.08 and 2.21 ± 0.04 mg g^?1 dry weight in leaves and callus, respectively. Apigenin, caffeic acid, p-coumaric acid, luteolin, rutin hydrate, vanillic acid, ferulic acid, salicylic acid, sinapic acid, and chlorogenic acid were detected by liquid chromatography–electrospray ionization multistage tandem mass spectrometry (LC-ESI-MS/MS) analysis in in vitro-grown leaves and callus tissue. Rutin hydrate, p-coumaric acid, and vanillic acid were found at approximately tenfold higher levels in callus than in leaves. This new micropropagation protocol, the first for P. armeniaca, could be used in industrial production for new herbal tea and germplasm conservation.     

CuO nanoparticles significantly influence in vitro culture,steviol glycosides,and antioxidant activities of <Emphasis Type="Italic">Stevia rebaudiana</Emphasis> Bertoni

Copper oxide (CuO) nanoparticles (NPs) synthesized through co-precipitation method were employed in MS media during in vitro culture of Stevia rebaudiana. Physiological characteristics, production of steviol glycosides, and antioxidative parameters were investigated in regenerated plants. CuO NPs had crystalline monoclinic cubic cuprous oxides with average size 47 nm. The NPs were applied at 0, 0.1, 1.0, 10, 100 and 1000 mg/L in MS media for direct organogenesis of S. rebaudiana from nodal segments. Shoot organogenesis was found highest (88.5%) at 10 mg/L CuO and average shoot length, mean number of shoot per explant, and fresh weight were also found significantly higher at the same concentration. High performance liquid chromatography (HPLC) illustrated significant rise of bioactive major steviol glycosides (rebaudioside A and stevioside) at 10 mg/L CuO NPs in MS media. The oxidative stress produced by CuO nanoparticles on S. rebaudiana was affirmed by antioxidant activities i.e. total antioxidant activity (TAC), total reducing power (TRP) and 2,2-diphenyl-1-picryl hydrazyl (DPPH)-free radical scavenging activity. The oxidative stress generated by NPs involved production of antioxidative molecules total phenolic content (TPC), total flavonoid content (TFC) depending on NPs concentration. The study concludes that copper oxide nanoparticles functions as a stimulator of bioactive components productions, and can be employed in in vitro batch cultures.     

Design,Synthesis, Antioxidant and Antibacterial Activities of Novel 2-((1-Benzyl-1<Emphasis Type="Italic">H</Emphasis>-1,2,3-Triazol-4-yl)methyl)-5-(2<Emphasis Type="Italic">H</Emphasis>Chromen- 3-yl)-2H-Tetrazoles

1,4-Disubstituted 1,2,3-triazole derivatives of 2H-chromene-3-tetrazoles synthesized regioselectively by copper(I)-catalyzed alkyne–azide cycloaddition (CuAAC) click reaction were characterized by ¹HNMR, ¹³C NMR, IR, and mass spectral data. These derivatives were screened for in vitro antioxidant activity using DPPH radical, H₂O₂ scavenging, and iron chelating activity methods and also evaluated for in vitro antibacterial activities against E. coli and S. aureus bacterial strains. The MIC and IC₅₀ values for all these compounds were found to match the docking scores and relevant binding energies with the receptor active sites. These results allows one to consider the compounds as leads for a new generation of antioxidant and antibacterial agents.     

Evaluation of antioxidant and antiproliferative metabolites of <Emphasis Type="Italic">Penicillium flavigenum</Emphasis> isolated from hypersaline environment: Tuz (Salt) Lake by Xcelligence technology

The aim of the study is the determination of antioxidant and antiproliferative activities of fungal isolates’ metabolites belonging to Penicillium flavigenum isolated from Lake Tuz, Turkey. Evaluation of the antioxidant activity, the total phenolic content and antiproliferative effect were evaluated with DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay, Folin-ciocalteu method, Xcelligence real-time cell analysis. The total phenolic content of these isolates were found 62–82 mg/GAE. Ethyl acetate extracts from identified isolates, P. flavigenum, showed cytotoxic effects on A549, MCF7, Caco-2 cell lines. IC₅₀ values of P. flavigenum ethyl acetate extracts were found 96.7 μg/mL for A549, 33.4 μg/mL for MCF7, 43.4 μg/mL for Caco-2 and 97.3 μg/mL for 3T3. Phenolic acids in the extracts from P. flavigenum were identified with HPLC and GC-MS. Penicillium flavigenum is a new report for Turkey. According to these findings, fungi-related secondary metabolites are very important sources in terms of antioxidant and antiproliferative effects.     

Isolation and characterization of anticancer flavone chrysin (5,7-dihydroxy flavone)-producing endophytic fungi from <Emphasis Type="Italic">Passiflora incarnata</Emphasis> L. leaves

Chrysin (5,7-dihydroxy flavone, ChR) is a flavone of plant origin, possessing numerous biomedical properties, such as antimicrobial, anti-inflammatory, anti-diabetic, anxiolytic, hepatoprotective, anti-aging and anticonvulsant activities. In this study, chrysin-producing fungal endophytes (A. alternata KT380662, C. capsici KT373967, and C. taiwanense PI-3 KX580307) were isolated from the leaves of Passiflora incarnata L. and characterised via morphology and internal transcribed spacer (ITS) sequences. Thin layer chromatography and high-performance liquid chromatography profiles of fungal extracts showed Rf values and retention times that closely match those of standard chrysin (ChR). Further, the production of fungal chrysin (FChR) was confirmed through UV-vis spectroscopy, FT-IR, LC-ESI-MS, and ¹H₁ NMR analysis. Among the isolated strains, A. alternata KT380662 was identified as having a high-level of ChR production, with rates measuring approximately 846 mg L^?1. On the other hand, in vitro anticancer and radical scavenging studies proved that FChR has significant cytotoxic activity against human liver carcinoma cells (HepG2). These results clearly imply that the isolated A. alternata KT380662 could serve as an alternative source for the commercial production of ChR, which holds anticancer and radical scavenging activities, and the fungal-derived ChR can be used in chemotherapy or in prodrug development.     

Phytochemical Analysis and Evaluation of the Antioxidant,Anti-Inflammatory,and Antinociceptive Potential of Phlorotannin-Rich Fractions from Three Mediterranean Brown Seaweeds

Phlorotannins, phenolic compounds produced exclusively by seaweeds, have been reported to possess various pharmacological properties. However, there have been few works on these compounds from Mediterranean seaweeds. In this study, we investigated the phytochemical analysis and pharmacological potential of phlorotannin-rich fractions from three brown seaweeds collected along the Tunisia coast: Cystoseira sedoides (PHT-SED), Cladostephus spongeosis (PHT-CLAD), and Padina pavonica (PHT-PAD). Phytochemical determinations showed considerable differences in total phenolic content (TPC) and phlorotannin content (PHT). The highest TPC level (26.45 mg PGE/g dry material (Dm)) and PHT level (873.14 μg PGE/g Dm) were observed in C. sedoides. The antioxidant properties of these three fractions assessed by three different methods indicated that C. sedoides displayed the highest total antioxidant activity among the three species (71.30 mg GAE/g Dm), as well as the free radical scavenging activity with the lowest IC₅₀ value in both DPPH (27.7 μg/mL) and ABTS (19.1 μg/mL) assays. Furthermore, the pharmacological screening of the anti-inflammatory potential of these fractions using in vivo models, in comparison to reference drugs, established a remarkable activity of PHT-SED at the dose of 100 mg/kg; the inhibition percentages of ear edema in mice model and paw edema in rats model were of 82.55 and 81.08%, respectively. The content of malondialdehyde (MDA) in liver tissues has been quantified, and PHT-SED was found to remarkably increase the lipid peroxidation in rat liver tissues. In addition, in two pain mice models, PHT-SED displayed a profound antinociceptive activity at 100 mg/kg and has proved a better analgesic activity when used in combination with the opioid drug, tramadol.     

Changes in phytochemical content and pharmacological activities of three <Emphasis Type="Italic">Chlorella</Emphasis> strains grown in different nitrogen conditions

The phytochemical content and biological activity of three Chlorella strains cultured in low (35 mg L^?1) or high (700 mg L^?1) nitrogen (N) and harvested on days 5 and 10 were evaluated. High N resulted in a higher biomass in Chlorella MACC 438 and MACC 452 while MACC 555 produced a higher biomass in low N. MACC 555 (low N/day 5) had the highest phenolic content, and MACC 438 in low N/day 5 and high N/day 5 accumulated the highest flavonoids and condensed tannins, respectively. Iridoids were most abundant in MACC 452 on low N/day 10. Harvest time had the greatest effect on the phytochemical content with phenolics, flavonoids, and condensed tannins decreasing over time and iridoids increasing in low N and decreasing in high N conditions. Extracts were more active in β-carotene-linoleic acid model compared to 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Most extracts had good antimicrobial activity. Extracts became less potent over time in the antioxidant, acetylcholinesterase inhibitory (AChE), and antimicrobial assays when growing in low N and more potent in the antioxidant and AChE assays when grown in high N. Thus, phytochemical content and biological activities of the three Chlorella strains were affected by N levels, harvest time, and strain.     

Synthesis,Antimicrobial and Antioxidant Activity of Pyrazole Based Sulfonamide Derivatives

A series of new sulfonamides have been synthesized from Ampyrone with different benzene sulfonyl chlorides to yield the N-1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl) benzenesulfonamides (4a–e). All synthesized compounds were characterized on the basis of FTIR, ¹H NMR, and ¹³C NMR, and also by the aid of mass spectral data. Further, all synthesized compounds have studied for their in vitro antimicrobial activities against selected bacterial as well as fungal strains by the agar well diffusion method. Free radical scavenging activity has been investigated by using DPPH method. Among all the synthesized compounds, 4b, 4d, and 4e exhibited significant antimicrobial and antioxidant activities.     

Biological activity of new flavonoid from <Emphasis Type="Italic">Hieracium pilosella</Emphasis> L.

Hieracium pilosella L. (Asteraceae) is a well-known plant used in ethno-medicine as its inflorescences are particularly rich in beneficial polyphenolics. This research aimed to elucidate the structure of a new flavone glycoside isolated from the inflorescences of Hieracium pilosella and evaluate its antioxidant, antimicrobial and antiproliferative activities. The chromatographic methods were successfully applied to isolate the new flavonoid. Its structure was determined by subsequent UV, NMR and MS experiments and identified as isoetin 4′-O-β-D-glucopyranoside. Free radical scavenging capacity was examined by measuring the scavenging activity of the new isoetin derivative on 2,2-diphenyl-1-picrylhydrazyl (DPPH). The compound was also screened for spectrum of antimicrobial activity using the agar well diffusion method. Minimum inhibitory concentration (MIC) for Pseudomonas aeruginosa ATCC 9027 was performed by the micro-dilution broth method. The antiproliferative effect of tested glycoside was assessed in two human tumor cell lines derived from lung (A549) and colon (HT-29) carcinoma and cell proliferation was determined by means of MTT method. The tested compound showed high antiradical activity, reducing the DPPH? with EC₅₀ 7.9 μM (3.7 µg/ml) and exhibited narrow antimicrobial spectrum among tested microorganisms. The compound was active against Pseudomonas aeruginosa ATCC 9027 (MIC 125 μg/ml) which is prone to causing infections that are difficult to treat due to it developing extremely rapid antibiotic resistance. In the antiproliferative studies, cell proliferation of the colon (HT-29) carcinoma cell line was significantly decreased after exposure to the compound. The results indicate that isoetin 4′-O-β-D-glucopyranoside possesses antioxidant capacity and very promising antibacterial activity and could have uses as an effective antipseudomonal agent as well a antiproliferative agent.     

Development and Evaluation of a Novel Delivery System Containing Phytophospholipid Complex for Skin Aging

Citrus auranticum and Glycyrrhiza glabra are rich in anti-oxidant polyphenols helpful in prevention of skin aging. Polyphenols have high polarity and lower skin penetration resulting in lower cutaneous delivery. The present work is attempted to develop a novel polyherbal phospholipid complex cream to improve cutaneous delivery of polyphenols for sustained anti-oxidant action. Phytochemical and in vitro anti-oxidant evaluation was done on methanolic extracts of orange peel and liquorice powder. Total phenolic content, total flavonoid content, and anti-oxidant assays were done on different ratios of orange peel and liquorice extract. Ratio 1:2 gave highest total phenolic content (TPC) (530.00?±?1.56 mg gallic acid equivalent (GAE)?g^?1 extract), total flavonoid content (TFC) (246.25?±?1.03 mg rutin equivalent (RUE)?g^?1 extract), 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity (87.99?±?0.64%), and H₂O₂ scavenging activity (72.47?±?0.86%) and hence was used for formulation. Solvent evaporation method using methanol with 1:1 extract to phospholipid ratio was found to have entrapment efficiency of 93.22?±?0.26%. Evaluation parameters like scanning electron microscopy (SEM), Fourier transform infrared spectrophotometry (FT-IR), and differential scanning calorimetry (DSC) confirmed formation of complex. The complex was formulated as oil-in-water cream and evaluated for various parameters. The optimized cream containing 1% complex was non-irritant and was found to be stable for 3-month period under conditions of stability study. Ex vivo diffusion studies showed that extract phospholipid complex cream had better retention of polyphenols in the skin when compared to conventional extract cream giving prolonged and stronger topical action. The cream had an anti-elastase activity of 28.02?±?0.95% at concentration of 3000 μg ml^?1 (w/v). Thus, the developed safe and stable polyherbal phytophospholipid complex cream exhibited good potential as anti-aging cosmeceutical.     

UV-B induces biomass production and nonenzymatic antioxidant compounds in three cyanobacteria

In the present study, the impact of low fluence rate of UV-B (0.045 W.m^?2) on biomass production, photosynthetic pigments (chlorophyll a, carotenoids, and phycobiliproteins), chlorophyll fluorescence, nonenzymatic antioxidants: proline, ascorbate, cysteine, and nonprotein thiols, total phenolic contents, and antioxidant potential (radical scavenging activity) was investigated in three cyanobacteria, viz. Nostoc muscorum, Phormidium foveolarum, and Arthrospira platensis. Selected fluence rate of UV-B caused enhancing effect on these parameters; however, the increased values of these attributes were greater in A. platensis followed by P. foveolarum and N. muscorum. Results indicate that UV-B (at selected fluence rate) could be used as technique that may modify cyanobacterial system for efficient and economic production of natural food supplements and/or natural pharmaceuticals.     

Characterization of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) lectin for biological activity

Lectins are proteins that are subject of intense investigations. Information on lectin from chickpea (Cicer arietinum L.) with respect to its biological activities are very limited. In this study, we purified lectin from the seeds of chickpea employing DEAE-cellulose and SP-Sephadex ion exchange chromatography and identified its molecular subunit mass as 35 kDa. The free radical scavenging activity of lectin measured by the DPPH assay has IC₅₀ of 0.88 µg/mL. Lectin exerted antifungal activity against Candida krusei, Fusarium oxysporium oxysporium, Saccharomyces cerevisiae and Candida albicans, while antibacterial activity against E. coli, B. subtilis, S. marcescens and P. aeruginosa. The minimum inhibitory concentrations were 200, 240, 160 and 140 µg for C. krusei, F. oxysporium, S. cerevisiae and C. albicans respectively. Lectin was further examined for its antiproliferative potential against cancerous cell line. The cell viability assay indicated a high inhibition activity on Ishikawa, HepG2, MCF-7 and MDA-MB-231 with IC₅₀ value of 46.67, 44.20, 53.58 and 37.46?µg/mL respectively. These results can provide a background for future research into the benefits of chickpea lectin to pharmacological perspective.     

Purification and activity characterization of polysaccharides in the medicinal lichen <Emphasis Type="Italic">Umbilicaria tornata</Emphasis> from Taibai Mountain,China

Water-soluble polysaccharides from Umbilicaria tornata (UTP) were purified and preliminarily characterized. The antioxidant and antitumor activities of crude UTP and two purified fractions (UTP-1 and UTP-2) were evaluated using in vitro experiments. The results showed that the molecular weights of UTP-1 and UTP-2 were 84.86 and 28.66 kDa, respectively. Both UTP-1 and UTP-2 were composed of glucose and xylose, with their molar ratios being 1.3:0.9 and 0.9:4.6, respectively. In addition, crude UTP, UTP-1 and UTP-2 showed dose-dependent DPPH and hydroxyl radical scavenging and reducing activities. However, crude UTP exhibited stronger antioxidant activity than UTP-1 and UTP-2, particularly in terms of DPPH radicals. Crude UTP and the two purified fractions inhibited the growth of HeLa, HepG2, A375, MCF-7, SGC7901 and Caco2 cancer cells in vitro. Compared with UTP-1 and UTP-2, crude UTP presented significantly higher antitumor activity in vitro against HeLa and HepG2 cells (p?<?0.05). These findings provide a scientific basis for the deeper exploration and resource development of U. tornata.     

Physiological,biochemical, antioxidant and growth characterizations of gibberellin and paclobutrazol-treated sweet leaf (<Emphasis Type="Italic">Stevia rebaudiana</Emphasis> B.) herb

A greenhouse experiment was designed to study the responses of Stevia rebaudiana herb to paclobutrazol (PBZ) and gibberellin (GA) treatments. GA and PBZ treatments caused no significant impact on photosynthesis pigments while they increased carbohydrates, amino acids and protein metabolites. Stevia showed a potent antioxidant activity through scavenging DPPH, NO^·; O ₂ ^·? and OH^· radicals which was highlighted in GA and PBZ treatments. The enzymatic and non-enzymatic antioxidant system of Stevia plant showed a significant increase in response to PBZ and GA treatments. PBZ treatment decreased plant growth while GA treatment had no significant effect on it. Collectively, both GA and PBZ treatments effectively increased metabolites and antioxidant property of Stevia herb.     

Alterations in the Antioxidant Status of Transgenic Roots of <Emphasis Type="Italic">Artemisia</Emphasis> spp. Representatives after <Emphasis Type="Italic">A. rhizogenes</Emphasis>-Mediated Genetic Transformation

The effect of A. rhizogenes-mediated genetic transformation on the antioxidant status of Artemisia tilesii, A. vulgaris, A. dracunculus, and A. annua transgenic roots has been studied. Antioxidant activity (AOA) of aqueous extracts was determined using methods based on the ability to reduce DPPH⁺ and ABTS⁺-radicals. The level of AOA (DPPH) in 50% of extracts obtained from transgenic roots was higher than the level of activity possessed by extracts from untransformed roots. An increased ability to reduce the ABTS+ radical was observed in 80% of the extracts. Extracts of A. annua and A. tilesii transgenic roots were the most active, while the lowest antioxidant activity was shown in A. dracunculus extracts. Thus, A. rhizogenes-mediated transformation has led to a change in the antioxidant status of the “hairy” roots of several Artemisia spp. plants (except A. vulgaris). It can be used as a method for the enhancement of the natural antiradical properties of plants belonging to the Artemisia genus.     

Production of an antioxidative peptide from hairy basil seed waste by a recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis>

Objective

To circumvent the time-consuming and costly problems associated with natural product extraction, a potential antioxidative peptide selected from hairy basil waste after oil extraction was produced by recombinant DNA technology.

Results

Because the target peptide is short, the recombinant peptide containing seven repeats of the target sequence, QTFQYSRGWTN, and the DNA fragment coding this sequence was cloned into the pQE-30 Xa expression vector and transformed into Escherichia coli. After 6 h of recombinant peptide expression in E. coli, the target peptide was purified by Ni²⁺ affinity chromatography and gel extraction. The expected 15 kDa recombinant target peptide construct was verified by modified dot blot analysis. Compared with the chemically synthesized peptide, the recombinant peptide revealed significantly higher antioxidant activities (p < 0.05), as determined by DPPH and ABTS radical scavenging assays, and in vitro DNA damage induced by hydroxyl radicals.

Conclusion

This approach provides an alternative to produce an antioxidative peptide that provides a potential scaffold for the further development of antioxidative peptides for industrial applications.

     

Impacts of methyl jasmonate and phenyl acetic acid on biomass accumulation and antioxidant potential in adventitious roots of <Emphasis Type="Italic">Ajuga bracteosa</Emphasis> Wall ex Benth., a high valued endangered medicinal plant

Ajuga bracteosa is a medicinally important plant globally used in the folk medicine against many serious ailments. In the present study, effects of two significant elicitors, methyl jasmonate (Me-J) and phenyl acetic acid (PAA) were studied on growth parameters, secondary metabolites production, and antioxidant potential in adventitious root suspension cultures of A. bracteosa. The results showed a substantial increase in biomass accumulation, exhibiting longer log phases of cultures growth in response to elicitor treatments, in comparison to control. Maximum dry biomass formation (8.88 DW g/L) was recorded on 32nd day in log phase of culture when  0.6 mg/L Me-J was applied; however, PAA at 1.2 mg/L produced maximum biomass (8.24 DW g/L) on day 40 of culture.  Furthermore, we observed the elicitors-induced enhancement in phenolic content (total phenolic content), flavonoid content (total flavonoid content) and antioxidant activity (free radical scavenging activity) in root suspension cultures of A. bracteosa. Application of 0.6 mg/L and 1.2 mg/L of Me-J, root cultures accumulated higher TPC levels (3.6 mg GAE/g DW) and (3.7 mg GAE/g DW) in the log phase and stationary phase, respectively, while 2.5 mg/L Me-J produced lower levels (1.4 mg GAE/g DW) in stationary phase of growth stages. Moreover, TFC and FRSA values were found in correspondence to TPC values in the respective growth phases at the similar elicitor treatment. Thus, a feasible protocol for establishment of adventitious roots in A. bracteosa was developed and enhancement in biomass and metabolite content in adventitious root was promoted through elicitation.     

Programmed cell death in the cyanobacterium <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> induced by allelopathic effect of submerged macrophyte <Emphasis Type="Italic">Myriophyllum spicatum</Emphasis> in co-culture system

This investigation demonstrates that programmed cell death (PCD) in a cyanobacterium, Microcystis aeruginosa, resulting from allelopathic stress induced by a submerged macrophyte, Myriophyllum spicatum, in a co-culture system. The hallmarks of PCD, caspase-3-like protease activity, DNA fragmentation, and destruction of cell ultrastructure, as well as intracellular PCD signaling radicals, reactive oxygen species (ROS), and nitric oxide (NO), were measured in M. aeruginosa cells co-cultured with M. spicatum for 7 days. The results showed a dose–response relationship between M. spicatum biomass and M. aeruginosa mortality. A caspase-3-like protease was activated and elevated from day 3. Thylakoid disintegration, cytoplasmic vacuolation, and fuzzy nuclear zone were observed by transmission electron microscopy, and distinct DNA fragmentation was detected in M. aeruginosa cells at a M. spicatum biomass of 6.0 g fresh weight (FW) L^?1 during the 7 days. Allelochemicals of total phenolic compounds (TPCs) were determined in co-culture water, and the concentration increased with increasing of M. spicatum biomass and co-culture time. Compared with the level of ROS production in the control group, a significant overproduction of ROS was detected in M. aeruginosa cells in the treatment group, and this was positively correlated with TPC concentration. Furthermore, the level of intracellular NO increased with the percent mortality of M. aeruginosa. The results indicated that a PCD pathway was induced in the cyanobacterium M. aeruginosa when co-cultured with the submerged macrophyte M. spicatum.