Heterologous production of hyaluronic acid in Nicotiana tabacum hairy roots expressing a human hyaluronan synthase 2

AbstractHyaluronic acid (HA), a unique polysaccharide with excellent Physico-chemical properties, is broadly used in pharmaceutical, biomedical, and cosmetic fields. It is widely present in all vertebrates, certain bacterial strains, and even viruses while it is not found in plants, fungi, and insects. HA is naturally synthesized by a class of integral membrane proteins called Hyaluronic acid synthase (HAS). Thus far, industrial production of HA is carried out based on either extraction from animal sources or large-scale microbial fermentation. The major drawbacks to using these systems are contamination with pathogens and microbial toxins. Recently, the production of HA through recombinant systems has received considerable attention. Plants are eco-friendly ideal expression systems for biopharmaceuticals production. In this study, the optimized human hyaluronic acid synthase2 (hHAS2) sequence was transformed into Nicotiana tabacum using Agrobacterium rhizogenes. The highest rhHAS2 concentration of 65.72 ng/kg (wet weight) in transgenic tobacco hairy roots was measured by the human HAS2 ELISA kit. The HA production in the transgenic hairy roots was verified by scanning electron microscope (SEM) and quantified by the HA ELISA kit. The DPPH radical scavenging activity of HA with the highest concentration of 0.56 g/kg (wet weight) showed a maximum activity of 46%. Gel Permeation Chromatography (GPC) analyses revealed the high molecular weight HA (HMW-HA) with about > 0.8 MDa.

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Heterologous production of Hyaluronic Acid in Nicotiana tabacum hairy roots expressing a human Hyaluronan synthase 2

10.21203/rs.3.rs-155813/v1 ◽

2021 ◽

Author(s):

arezoo nazeri ◽

Ali Niazi ◽

Alireza Afsharifar ◽

Seyed Mohsen Taghavi ◽

Ali Moghadam ◽

...

Keyword(s):

Hyaluronic Acid ◽

Nicotiana Tabacum ◽

Hairy Roots ◽

Large Scale ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Gel Permeation Chromatography ◽

Maximum Activity ◽

Bacterial Strains ◽

Wet Weight

Abstract Hyaluronic acid (HA), a unique polysaccharide with excellent physicochemical properties, is broadly used in pharmaceutical, biomedical, and cosmetic fields. It is widely present in all vertebrates, certain bacterial strains, and even viruses while it is not found in plants, fungi, and insects. HA is naturally synthesized by a class of integral membrane proteins called Hyaluronic acid synthase (HAS). Thus far, industrial production of HA is carried out based on either extraction from animal sources or large-scale microbial fermentation. The major drawbacks to using these systems are contamination with pathogens and microbial toxins. Recently, the production of HA through recombinant systems has received considerable attention. Plants are eco-friendly ideal expression systems for biopharmaceuticals production. In this research, the optimized human hyaluronic acid synthase2 (hHAS2) sequence was transformed into Nicotiana tabacum using Agrobacterium rhizogenes. The highest hHAS2 production in the tobacco hairy roots was 65.72 ng/kg (wet weight). The extracted HA was verified and quantified by the HA ELISA kit. The DPPH radical scavenging activity of HA with the highest concentration of 0.56 g/kg (wet weight) showed the maximum activity of 46%. Gel Permeation Chromatography (GPC) analyses revealed the high molecular weight HA with about > 0.8 MDa.

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Antimicrobial and antioxidant potentials of non-cytotoxic extracts of corticolous lichens sampled in Armenia

AMB Express ◽

10.1186/s13568-021-01271-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Razmik Sargsyan ◽

Arsen Gasparyan ◽

Gohar Tadevosyan ◽

Hovik Panosyan

Keyword(s):

Antifungal Activity ◽

Methanol Extract ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Cytotoxic Activities ◽

Bacterial Strains ◽

Evernia Prunastri ◽

Wide Range ◽

Antibacterial And Antifungal ◽

Acetone Extracts

AbstractDue to wide range of secondary metabolites, lichens were used from antiquity as sources of colorants, perfumes and medicaments. This research focuses on exploring the antioxidant, antimicrobial and cytotoxic activities of methanol, ethanol, acetone extracts and aqueous infusions of corticolous lichens sampled from Armenia. Methanol, ethanol and acetone extracts from all tested lichens were active against Gram-positive bacterial strains. The most effective solvent to retrieve antimicrobial compounds was methanol. Aqueous infusions of tested lichens didn’t show any significant antibacterial and antifungal activity. The highest antimicrobial activity was observed for methanol extract of Ramalina sinensis. The minimum inhibitory concentration of methanol extract of Ramalina sinensis were 0.9–1.8 mg mL− 1. Pseudevernia furfuracea demonstrated antifungal activity (Ø 12 mm). Methanol extract of Parmelia sulcata demonstrated largest 1,1-diphenyl-2-picryl-hydrazil (DPPH) radical scavenging activity (71 %). The cytotoxicity was measured on human HeLa (cervical carcinoma) cell lines using microculture tetrazolium test assay. The IC50 values estimated for methanol extracts of Peltigera praetextata, Evernia prunastri, Ramalina sinensis and Ramalina farinacea species in HeLa cell line were within 1.8–2.8 mg mL− 1 and considered as non-cytotoxic. Obtained results suggest that studied lichens can be prospective in biotechnologies as alternative sources of antimicrobial and antioxidant substances.

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Aktywność przeciwutleniająca i przeciwbakteryjna miodu, propolisu oraz pyłku kwiatowego

Postępy Fitoterapii ◽

10.25121/pf.2020.21.3.123 ◽

2020 ◽

Vol 21 (3) ◽

Author(s):

Magdalena Woźniak ◽

Lucyna Mrówczyńska ◽

Anna Sip ◽

Marta Babicka ◽

Tomasz Rogoziński ◽

...

Keyword(s):

Antioxidant Activity ◽

Phenolic Compounds ◽

Pathogenic Bacteria ◽

Radical Scavenging Activity ◽

Colorimetric Method ◽

Antioxidant Properties ◽

Radical Scavenging ◽

Total Content ◽

Diffusion Method ◽

Bacterial Strains

Introduction. Honey, propolis and pollen belong to bee products that have beneficial biological properties. These products exhibit e.g. antibacterial, antifungal and antioxidant properties. Due to biological activity and natural origin, bee products are used, e.g. in the food industry, cosmetology and pharmacy. Aim. The aim of the study was to compare the antioxidant and antibacterial activity of honey, propolis and pollen from an apiary located in Wielkopolska Province. Material and methods. Honey, propolis and pollen used in this study came from the same apiary located in Wielkopolska Province. The antioxidant potential of bee products was evaluated applying DPPH· free radical scavenging activity assay. The antimicrobial activity of the tested bee products was determined by the point-diffusion method against 13 strains of pathogenic and potentially pathogenic bacteria. In addition, the total content of phenolic compounds in honey, propolis and pollen was determined by the colorimetric method. Results. Propolis exhibited higher antioxidant activity, in comparison to honey and pollen. The antiradical activity of propolis was equal to 80% approx. activity of Trolox, the standard antioxidant. Among tested bee products, propolis was characterized by the highest total phenols content. In addition, honey, propolis and pollen showed antagonistic activity against tested bacterial strains. Conclusions. The obtained results indicate that among the tested bee products of native origin, i.e. honey, propolis and pollen, propolis characterized by the highest antioxidant activity and the total content of phenolic compounds. In addition, all bee products showed bactericidal activity against the tested bacterial strains.

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Bioprocessing of Squid Pens Waste into Chitosanase by Paenibacillus sp. TKU047 and Its Application in Low-Molecular Weight Chitosan Oligosaccharides Production

Polymers ◽

10.3390/polym12051163 ◽

2020 ◽

Vol 12 (5) ◽

pp. 1163 ◽

Cited By ~ 2

Author(s):

Chien Thang Doan ◽

Thi Ngoc Tran ◽

Van Bon Nguyen ◽

Trung Dung Tran ◽

Anh Dzung Nguyen ◽

...

Keyword(s):

Biological Activities ◽

Radical Scavenging Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Radical Scavenging ◽

Maximum Activity ◽

Degree Of Polymerization ◽

Chitosan Oligosaccharide ◽

Scavenging Activity ◽

Paenibacillus Sp

Chitosan oligosaccharide (COS) has become of great interest in recent years because of its worthy biological activities. This study aims to produce COS using the enzymatic method, and investigates Paenibacillus sp. TKU047, a chitinolytic-producing strain, in terms of its chitosanase productivity on several chitinous material-containing mediums from fishery process wastes. The highest amount of chitosanase was produced on the medium using 2% (w/v) squid pens powder (0.60 U/mL) as the single carbon and nitrogen (C/N) source. The molecular mass of TKU047 chitosanase, which could be the smallest one among chitinases/chitosanases from the Paenibacillus genus, was approximately 23 kDa according to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. TKU047 chitosanase possessed the highest activity at 60 °C, pH 7, and toward chitosan solution with a higher degree of deacetylation (DDA) value. Additionally, the hydrolysis products of 98% DDA chitosan catalyzed by TKU047 chitosanase showed the degree of polymerization (DP) ranging from 2 to 9, suggesting that it was an endo-type activity chitosanase. The free radical scavenging activity of the obtained chitosan oligosaccharide (COS) was determined. The result showed that COS produced with Paenibacillus sp. TKU047 chitosanase expressed a higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity than that from the commercial COSs with maximum activity and IC50 values of 81.20% and 1.02 mg/mL; 18.63% and 15.37 mg/mL; and 15.96% and 15.16 mg/mL, respectively. As such, Paenibacillus sp. TKU047 may have potential use in converting squid pens waste to produce chitosanase as an enzyme for bio-activity COS preparation.

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Probiotic and Antioxidant Potential of Lactobacillus reuteriLR12 and Lactobacillus lactisLL10 Isolated from Pineapple Puree and Quality Analysis of Pineapple-Flavored Goat Milk Yoghurt during Storage

Microorganisms ◽

10.3390/microorganisms8101461 ◽

2020 ◽

Vol 8 (10) ◽

pp. 1461 ◽

Cited By ~ 3

Author(s):

Naif Abdullah Al-Dhabi ◽

Mariadhas Valan Arasu ◽

Ponnuswamy Vijayaraghavan ◽

Galal Ali Esmail ◽

Veeramuthu Duraipandiyan ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Radical Scavenging Activity ◽

Starter Culture ◽

Radical Scavenging ◽

Reducing Power ◽

Goat Milk ◽

Quality Analysis ◽

Scavenging Activity ◽

Bacterial Strains ◽

Adhesion Properties

In recent years, studies have focused on the therapeutic properties of probiotics to eliminate pathogenic microorganisms associated with various diseases. Lactobacilli are important probiotics groups that have been found to possess many health-promoting activities. This study was carried out to isolate LactobacillusreuteriLR12 and L. lactisLL10 from pineapple puree. The invitro analysis to evaluate probiotic characteristics of the isolated bacteria included survival in bile and acid tolerance. The cell-free supernatant of L. reuteri LR12 was effective against various pathogenic bacteria and fungi compared with L. lactisLL10. These two bacterial strains have strong anti-biofilm activity (100%) against Enterococcus faecalis, Staphylococcus aureus, and Bacillus cereus. The bacterial strains exhibited adhesion properties to HT-29 cells (human colorectal adenocarcinoma). These bacteria showed DPPH- (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical scavenging activity, scavenging of hydroxyl radical activity, superoxide radical scavenging activity, and reducing power activity in the range of 72% ± 3%to 89.3% ± 1.7%, 64% ± 2.7%to 66.8% ± 1.5%, 59.8% ± 4.1% to 63.8% ± 2.1%, and 60.4% ± 1.8%to 66.1% ± 3.3%, respectively. Pineapple puree was used as the starter culture with milk for 2 days for yogurt preparation. Pineapple puree increased flavor and showed the physicochemical properties of yogurt. The finding of the sensory evaluation revealed no significant change compared with the control, except the appearance of yogurt. These findings show that Lactobacilli and pineapple puree have potential use in various probiotic preparations for the fermentation industry.

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Evaluation of Antioxidant Capacity and Antiproliferative Activity of Fruit Extract of Dry Figs (Ficus carica L.) on MDA MB-468 Cell Line

10.21203/rs.3.rs-1145771/v1 ◽

2021 ◽

Author(s):

Anurag Maurya ◽

Eepsita Priyadarshini ◽

Paulraj Rajamani

Keyword(s):

Antiproliferative Activity ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Maximum Activity ◽

Ficus Carica ◽

Total Phenolic ◽

Fruit Extract ◽

Polar Solvents ◽

Dietary Component

Abstract Fig (Ficus carica L.; Hindi: Anjeer) is one of the the earliest domesticated fruit. Present study investigates its antiproliferative activity against breast cancer (MDA MB-468) cells. Dry figs were mascerated in sixteen solvent-combinations of varying polartiy and extracts were assessed for antiproliferative activity in vitro. Extracts of moderatly polar solvents, particularly 80% aqueous-methanol extract showed maximum activity (IC50 = 17.9 mg IDF/ml). Antiproliferative efficacy of extracts was supported by respective total phenolic content (41-236 mg GAE/100g) and various antioxidant capacities; ABTS (14-354 µM TE/g), DPPH (18-460 µM TE/g), FRAP (28.7-66.5 µM FeSO4 equivalents/g), ORAC (7.9-32.8 µM TE/g), and superoxide radical scavenging activity. Flow cytometry analysis revealed that extracts of lower IC50 induce more cell death. Phenolic compounds (cyanidin 3-O-glucoside, quercetin 3-β-glucoside, rutin, trans-cinnamic acid, ellagic acid and ferulic acid) were analysed by UPLC-PDA technique and their concentrations in extracts were found to be negatively correlated with respective IC50 values. The negative correlation suggests the involvement of phenolics in fig’s antiproliferative mechanism. Therefore, the study advocates fig as a dietary component having anticancer potential.

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Immobilization of Chaga extract in alginate beads for modified release: Simplicity meets efficiency

Hemijska industrija ◽

10.2298/hemind190819028p ◽

2019 ◽

Vol 73 (5) ◽

pp. 325-335

Author(s):

Predrag Petrovic ◽

Katarina Ivanovic ◽

Charly Octrue ◽

Mica Tumara ◽

Aleksandra Jovanovic ◽

...

Keyword(s):

Radical Scavenging Activity ◽

Radical Scavenging ◽

Entrapment Efficiency ◽

Gastrointestinal Diseases ◽

Hot Water ◽

Parasitic Fungus ◽

Prolonged Release ◽

Bacterial Strains ◽

Active Metabolites ◽

Inonotus Obliquus

Chaga (Inonotus obliquus) is a parasitic fungus, which has been used in traditional medicine in Russia and other northern European countries in the treatment of cancer, gastrointestinal andliver diseases. It has been a subject of intensive researchrecently, confirming many of its health-beneficial effects. In order to obtain a product that would allow modified and prolonged release of the Chaga?s active metabolites, hot water Chaga extract was immobilized using calcium - alginate. The extract, which was predominantly composed of carbohydrates (57 %), also contained a relatively high amount of antioxidants/phenolic compounds (130 mg gallic acid equivalents per g of dry extract) and exhibited pronounced radical scavenging activity. It showed significant antibacterial activity as well, inhibiting growth of tested bacterial strains at concentrations of 1.25-20 mg/mL. Entrapment efficiency was about 80 %, and the extract-alginate system showed pH-dependant extract release; there was negligible release at pH 1.75 (gastric pH), and the release gradually increased with the increase in pH, reaching ~43 % of immobilized extract at pH 8.5 after 90 min. Such a product could be used as a dietary supplement, adjuvant in therapy of gastrointestinal diseases or as a food additive.

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IN VITRO ANTIOXIDANT AND ANTIHYPERGLYCEMIC ACTIVITIES OF HYDROETHANOLIC LEAF EXTRACT OF ACACIA CATECHU (L.F) WILLD

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i11.20299 ◽

2017 ◽

Vol 10 (11) ◽

pp. 190

Author(s):

Ramesh Balu ◽

Jayabharathi V

Keyword(s):

Leaf Extract ◽

Antioxidant Activities ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Pharmaceutical Preparations ◽

Maximum Activity ◽

Increase With Increase ◽

Nitric Oxide Radical ◽

Acacia Catechu

Objective: The aim of this study was to analyze the antioxidant and antihyperglycemic activities of hydroethanolic leaf extract of Acacia catechu (L.f) Willd. Methods: The A. catechu (L.f) Willd. leaf extract was subjected to antioxidant activities such as 1,1-diphenyl-1-picryl hydrazyl (DPPH), hydroxyl radical (OH−), nitric oxide radical (NO−), and superoxide radical (SO) scavenging assays. Further, the leaf extract was subjected to antihyperglycemic activity by α-amylase inhibition assay.Results: The free radical scavenging activity of the plant extract was found to increase with increase in concentration (20-100 μg/ml) and the maximum activity was obtained at 100 μg/ml, which was compared with the standard ascorbic acid. The percentage inhibition of free radicals was found to be 62.94±0.91%, 63.53±1.08%, 71.04±0.48%, and 51.50±0.86% for DPPH, OH−, NO−, and SO−, respectively. The IC50 for DPPH, OH−, NO−, and SO were recorded as 73.81±0.95, 69.03±0.58, 60.74±0.37, and 83.10±1.00 μg/ml. The IC50 value of α-amylase inhibitory activity was found to be 71.17 μg/ml.Conclusion: The present investigations suggest that the A. catechu (L.f) Willd may serve as a potential source of natural antioxidant and antihyperglycemic agent that could have great importance in pharmaceutical preparations.

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Ectopic Expression of Production of Anthocyanin Pigment 1 (PAP1) Improves the Antioxidant and Anti-Melanogenic Properties of Ginseng (Panax ginseng C.A. Meyer) Hairy Roots

Antioxidants ◽

10.3390/antiox9100922 ◽

2020 ◽

Vol 9 (10) ◽

pp. 922

Author(s):

Sora Jin ◽

Tae Kyung Hyun

Keyword(s):

Panax Ginseng ◽

Hairy Roots ◽

Biological Activities ◽

Radical Scavenging Activity ◽

Differentially Expressed Gene ◽

Ectopic Expression ◽

Radical Scavenging ◽

Ginseng Root ◽

Anthocyanin Pigment ◽

Anthocyanin Production

The development of genetically engineered cell cultures has been suggested as a potential approach for the production of target compounds from medicinal plants. In this study, we generated PAP1 (production of anthocyanin pigment 1)-overexpressing ginseng (Panax ginseng C.A. Meyer) hairy roots to improve the production of anthocyanins, as well as the bioactivity (e.g., antioxidant and whitening activities) of ginseng. Based on differentially expressed gene analysis, we found that ectopic expression of PAP1 induced the expression of genes involved in the ‘phenylpropanoid biosynthesis’ (24 genes), and ‘flavonoid biosynthesis’ (17 genes) pathways, resulting in 191- to 341-fold increases in anthocyanin production compared to transgenic control (TC) hairy roots. Additionally, PAP1-overexpressing ginseng hairy roots exhibited an approximately seven-fold higher DPPH-free radical scavenging activity and 10-fold higher ORAC value compared to the TC. In α-melanocyte-stimulating hormone-stimulated B16F10 cells, PAP1-overexpressing ginseng hairy roots strongly inhibited the accumulation of melanin by 50 to 59% compared to mock-control. Furthermore, results obtained by quantitative real-time PCR, western blot, and tyrosinase inhibition assay suggested that the anti-melanogenic activity of PAP1-overexpressing ginseng hairy roots is mediated by tyrosinase activity inhibition. Taken together, our results suggested that the ectopic expression of PAP1 is an effective strategy for the enhancement of anthocyanin production, which improves the biological activities of ginseng root cultures.

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Isolation of phytoconstituents and evaluation of biological potentials of Berberis hispanica from Algeria

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v13i2.36133 ◽

2018 ◽

Vol 13 (2) ◽

pp. 179 ◽

Cited By ~ 2

Author(s):

Redouane Lemoui ◽

Samira Benyahia ◽

Labib Noman ◽

Ilham Bencherchar ◽

Feyza Oke-Altuntas ◽

...

Keyword(s):

Biological Activities ◽

Radical Scavenging Activity ◽

Video Clip ◽

Radical Scavenging ◽

2D Nmr ◽

Butylated Hydroxytoluene ◽

Klebsiella Pneumonia ◽

Bacterial Strains ◽

Remarkable Effect ◽

Berberis Hispanica

The aim of this study was to isolate the phytoconstituents and evaluate the biological activities of Berberis hispanica. Three phenolic compounds (tamarixetin, caffeic acid and rutin) were isolated from B. hispanica. The structures of the pure compounds were elucidated by spectroscopic and mass-spectrometric analyses, including 1D-, 2D-NMR, and HPLC-TOF/MS. In addition, antimicrobial, anti-oxidant and anti-proliferative effects of the extracts, some fractions and isolated compounds were evaluated. The extracts of B. hispanica were evaluated against six bacterial strains and exhibited the highest activity against Klebsiella pneumonia (15 mm at 100 mg/mL). Fraction T36 (IC50<5 μg/mL) from the n-butanol extract displayed higher radical scavenging activity than butylated hydroxytoluene. The isolated compounds were evaluated for their antiproliferative effects against human cervical adenocarcinoma (HeLa) cell line by real time cell analyzer assay and tamarixetin exhibited the remarkable effect (IC50<50 μg/mL) on HeLa cells. This study supports the documented medicinal effects of B. hispanica.Video Clip of Methodology:Antiproliferative Assay: 4 min 31 sec <a href="https://www.youtube.com/v/vi_fS47fpY0&t=3s">Full Screen</a> <a href="https://www.youtube.com/watch?v=vi_fS47fpY0&t=3s">Alternate</a>

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