scholarly journals PRODUCTION, PURIFICATION AND INHIBITION OF ALGINATE LY-ASE FROM LOCAL ISOLATE OF PSEUDOMONAS AERUGINOSA NA11

2020 ◽  
Vol 51 (6) ◽  
pp. 1726-1739
Author(s):  
R. W. Younis
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antioxidant Activity ◽  
Incubation Temperature ◽  
Specific Activity ◽  
Radical Scavenging ◽  
Deae Cellulose ◽  
High Specific Activity ◽  
Alginate Lyase ◽  
Maximum Efficiency ◽  
Local Isolate

The point of this study was for determine of the optimum conditions and purification of alginate lyase from local isolate of Pseudomonas aeruginosa NA11, and inhibition of enzyme by various plant extracts. Forty local  isolate s of pathogenic Pseudomonas aeruginosa were screened for their ability to produce alginate lyase. Local isolate of P.aeruginosa NA11 showed the maximum efficiency for produce of alginate lyase with high specific activity (14.4 U/mg). Several factors that influence on alginate lyase production from local isolate of P.aeruginosa NA11 were studied, these factors included the type of media, carbon source, nitrogen source, the incubation temperature, pH, and the incubation period. The highest yield of alginate lyase was obtained with the A medium supplemented with 0.5 % of glucose and sodium nitrate at pH 7.5 after 24 hr. incubation at 37 °C. Two chromatographic techniques were used for purification of alginate lyase after precipitation by ammonium sulphate with saturated ratio (0-70 %),  including, ion exchange chromatography by DEAE-cellulose and gel filtration by Sephadex G-100. The two steps gave the specific activity of 155.8 U/mg protein, the purification fold was 4.15 and enzymatic yield was 64 %. The molecular weight of partial purified alginate lyase was 57 KDa. The results of antioxidant activity tests for different plants extracts utilizing DPPH radical scavenging activity were showed that the saad extract has high antioxidant activity (71 %) more than other plants extracts. While the results for inhibition experiment of alginate lyase were demonstrated that saad extract was the best inhibitor with inhibition ratio of 86 %.

Folic acid compounds in romaine lettuce

1977 ◽  
Vol 55 (8) ◽  
pp. 865-868 ◽  
Author(s):  
K. K. Batra ◽  
J. R. Wagner ◽  
E. L. R. Stokstad
Keyword(s):  
Folic Acid ◽  
Specific Activity ◽  
Deae Cellulose ◽  
High Specific Activity ◽  
Microbiological Assay ◽  
Romaine Lettuce ◽  
Positive Identification

The composition of folate coenzymes in romaine lettuce was studied. Lettuce extract was purified on QAE-Sephadex A-25 and folate compounds were separated into a monoglutamate fraction and a polyglutamate fraction by chromatography on Sephadex G-15. Both the mono- and poly-glutamate fractions were resolved on DEAE-cellulose. Positive identification of DEAE peaks was made by further cochromatography with high specific activity radioactive marker folate compounds and with differential microbiological assay. The distribution of folate compounds in lettuce is as follows: 32% 5-CH3-H4PteGlu; 1% 5-CHO-H4PteGlu; 3% 5-CHO-H4PteGlu4; 9% 5-CH3-H4PteGlu4; 13% 5-CHO-H4PteGlu5; and 31% 5-CH3-H4PteGlu5.

scholarly journals Antioxidant Activity and Cytotoxicity against Cancer Cell Lines of the Extracts from Novel Xylaria Species Associated with Termite Nests and LC-MS Analysis

Antioxidants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1557
Author(s):  
Niwana Wangsawat ◽  
Lutfun Nahar ◽  
Satyajit D. Sarker ◽  
Cherdchai Phosri ◽  
Andrew R. Evans ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Cell Lines ◽  
Total Phenolic Content ◽  
Phenolic Content ◽  
Specific Activity ◽  
Antioxidant Activities ◽  
Radical Scavenging ◽  
Bioactive Metabolites ◽  
Total Phenolic ◽  
Ic50 Values

Xylaria species associated with termite nests or soil have been considered rare species in nature and the few which have been reported upon have been found to act as a rich source of bioactive metabolites. This study evaluated 10 ethyl acetate extracts of five new Xylaria species associated with termite nests or soil for their antioxidant activity, and cytotoxicity against different cancer and normal cell lines. DPPH and ABTS radical scavenging activities of the extracts demonstrated strong capacity with low IC50 values. The highest observed activities belonged to X. vinacea SWUF18-2.3 having IC50 values of 0.194 ± 0.031 mg/mL for DPPH assay and 0.020 ± 0.004 mg/mL for ABTS assay. Total phenolic content ranged from 0.826 ± 0.123 to 3.629 ± 0.381 g GAE/g crude extract which correlated with antioxidant activities. The high total phenolic content could contribute to the high antioxidant activities. Cytotoxicity was recorded against A549, HepG2, HeLa and PNT2 and resulted in broad spectrum to specific activity depending on the cell lines. The highest activities were observed with X. subintraflava SWUF16-11.1 which resulted in 11.15 ± 0.32 to 13.17 ± 2.37% cell viability at a concentration of 100 µg/mL. Moreover, LC-MS fingerprints indicated over 61 peaks from all isolates. There were 18 identified and 43 unidentified compounds compared to mass databases. The identified compounds were from various groups of diterpenoids, diterpenes, cytochalasin, flavones, flavonoids, polyphenols, steroids and derivatives, triterpenoids and tropones. These results indicate that Xylaria spp. has abundant secondary metabolites that could be further explored for their therapeutic properties.

scholarly journals Purification of pig synovial collagenase to high specific activity

1979 ◽  
Vol 183 (3) ◽  
pp. 647-656 ◽  
Author(s):  
T E Cawston ◽  
J A Tyler
Keyword(s):  
Tissue Culture ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
High Specific Activity ◽  
Polyacrylamide Gel ◽  
Active Enzyme ◽  
Latent Form

1. Pig synovium in tissue culture secretes a specific collagenase in a latent form. 2. The latent enzyme was concentrated by (NH4)2SO4 precipitation and activated with 4-aminophenylmercuric acetate, and the active enzyme was purified by chromatography on Ultrogel AcA44, DEAE-cellulose, heparin-Sepharose and a zinc-chelate medium to a specific activity of 53 400 units/mg. of protein. 3. The enzyme was shown to be essentially homogeneous by polyacrylamide-gel electrophoresis. 4. The purified collagenase digested collagen to give the characteristic three-quarter and one-quarter pieces.

scholarly journals Dominant Role of Paraoxonases in Inactivation of the Pseudomonas aeruginosa Quorum-Sensing Signal N-(3-Oxododecanoyl)-l-Homoserine Lactone

2008 ◽  
Vol 76 (6) ◽  
pp. 2512-2519 ◽  
Author(s):  
John F. Teiber ◽  
Sven Horke ◽  
Donovan C. Haines ◽  
Puneet K. Chowdhary ◽  
Junhui Xiao ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Specific Activity ◽  
Dominant Role ◽  
High Specific Activity ◽  
Homoserine Lactone ◽  
Mouse Lung ◽  
Calcium Dependent ◽  
Protein Levels ◽  
Human Sera

ABSTRACT The pathogenic bacterium Pseudomonas aeruginosa causes serious infections in immunocompromised patients. N-(3-Oxododecanoyl)-l-homoserine lactone (3OC12-HSL) is a key component of P. aeruginosa's quorum-sensing system and regulates the expression of many virulence factors. 3OC12-HSL was previously shown to be hydrolytically inactivated by the paraoxonase (PON) family of calcium-dependent esterases, consisting of PON1, PON2, and PON3. Here we determined the specific activities of purified human PONs for 3OC12-HSL hydrolysis, including the common PON1 polymorphic forms, and found they were in the following order: PON2 ≫ PON1192R > PON1192Q > PON3. PON2 exhibited a high specific activity of 7.6 ± 0.4 μmols/min/mg at 10 μM 3OC12-HSL, making it the best PON2 substrate identified to date. By use of class-specific inhibitors, approximately 85 and 95% of the 3OC12-HSL lactonase activity were attributable to PON1 in mouse and human sera, respectively. In mouse liver homogenates, the activity was metal dependent, with magnesium- and manganese-dependent lactonase activities comprising 10 to 15% of the calcium-dependent activity. In mouse lung homogenates, all of the activity was calcium dependent. The calcium-dependent activities were irreversibly inhibited by extended EDTA treatment, implicating PONs as the major enzymes inactivating 3OC12-HSL. In human HepG2 and EA.hy 926 cell lysates, the 3OC12-HSL lactonase activity closely paralleled the PON2 protein levels after PON2 knockdown by small interfering RNA treatment of the cells. These findings suggest that PONs, particularly PON2, could be an important mechanism by which 3OC12-HSL is inactivated in mammals.

scholarly journals Phospholipase A2 enzyme from the venom of Egyptian honey bee Apis mellifera lamarckii with anti-platelet aggregation and anti-coagulation activities

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Doaa A. Darwish ◽  
Hassan M. M. Masoud ◽  
Mohamed M. Abdel-Monsef ◽  
Mohamed S. Helmy ◽  
Hind A. Zidan ◽  
...  
Keyword(s):  
Apis Mellifera ◽  
Platelet Aggregation ◽  
Phospholipase A2 ◽  
Honey Bee ◽  
Specific Activity ◽  
Deae Cellulose ◽  
High Specific Activity ◽  
Inhibitory Effect ◽  
Bee Venom ◽  
Very High

Abstract Background Honey bee venom contains various enzymes with wide medical and pharmaceutical applications. Results The phospholipase A2 (PLA2) has been apparently purified from the venom of Egyptian honey bee (Apis mellifera lamarckii) 8.9-fold to a very high specific activity of 6033 U/mg protein using DEAE–cellulose and Sephacryl S-300 columns. The purified bee venom PLA2 is monomeric 16 kDa protein and has isoelectric point (pI) of 5.9. The optimal activity of bee venom PLA2 was attained at pH 8 and 45 °C. Cu2+, Ni2+, Fe2+, Ca2+, and Co2+ exhibited a complete activating effect on it, while Zn2+, Mn2+, NaN3, PMSF, N-Methylmaleimide, and EDTA have inhibitory effect. Conclusions The purified bee venom PLA2 exhibited anti-platelet aggregation and anti-coagulation activities which makes it promising agent for developing novel anti-clot formation drugs in future.

Effects of Heparin Oligosaccharides with High Affinity for Antithrombin III in Experimental Venous Thrombosis

1982 ◽  
Vol 47 (03) ◽  
pp. 244-248 ◽  
Author(s):  
D P Thomas ◽  
Rosemary E Merton ◽  
T W Barrowcliffe ◽  
L Thunberg ◽  
U Lindahl
Keyword(s):  
Antithrombin Iii ◽  
Specific Activity ◽  
High Specific Activity ◽  
Factor Xa ◽  
Blood Levels ◽  
Thrombin Time ◽  
High Affinity ◽  
Very High

SummaryThe in vitro and in vivo characteristics of two oligosaccharide heparin fragments have been compared to those of unfractionated mucosal heparin. A decasaccharide fragment had essentially no activity by APTT or calcium thrombin time assays in vitro, but possessed very high specific activity by anti-Factor Xa assays. When injected into rabbits at doses of up to 80 ¼g/kg, this fragment was relatively ineffective in impairing stasis thrombosis despite producing high blood levels by anti-Xa assays. A 16-18 monosaccharide fragment had even higher specific activity (almost 2000 iu/mg) by chromogenic substrate anti-Xa assay, with minimal activity by APTT. When injected in vivo, this fragment gave low blood levels by APTT, very high anti-Xa levels, and was more effective in preventing thrombosis than the decasaccharide fragment. However, in comparison with unfractionated heparin, the 16-18 monosaccharide fragment was only partially effective in preventing thrombosis, despite producing much higher blood levels by anti-Xa assays.It is concluded that the high-affinity binding of a heparin fragment to antithrombin III does not by itself impair venous thrombogenesis, and that the anti-Factor Xa activity of heparin is only a partial expression of its therapeutic potential.

Autoprothrombin C Activity from Prothrombin with Snake Venom or Trypsin

1962 ◽  
Vol 08 (03) ◽  
pp. 425-433 ◽  
Author(s):  
Ewa Marciniak ◽  
Edmond R Cole ◽  
Walter H Seegers
Keyword(s):  
Snake Venom ◽  
Thrombolytic Agent ◽  
Sodium Citrate ◽  
Specific Activity ◽  
High Specific Activity ◽  
Citrate Solution ◽  
Clotting Factors ◽  
Activation Products ◽  
Russell’S Viper ◽  
Autoprothrombin C

SummarySuitable conditions were found for the generation of autoprothrombin C from purified prothrombin with the use of Russell’s viper venom or trypsin. DEAE chromatographed prothrombin is structurally altered and has never been found to yield autoprothrombin C and also did not yield it when Russell’s viper venom or trypsin were used. Autoprothrombin C is derived from prothrombin with tissue extract thromboplastin, but not in large amounts with the intrinsic clotting factors. With the latter thrombin and autoprothrombin III are the chief activation products. Autoprothrombin III concentrates were prepared from serum and upon activation with 25% sodium citrate solution or with Russell’s viper venom large amounts of autoprothrombin C were obtained, and this was of high specific activity. Theoretically trypsin is not a thrombolytic agent, but on the contrary should lead to intravascular clotting.

Evaluation of antioxidant and Anthelmintic activity of methanolic flower extract of Nyctanthes arbor-tristis

2018 ◽  
Vol 9 (1) ◽  
pp. 78
Author(s):  
Vijaya Jyothi M ◽  
Bhargav E ◽  
Pavan Kumar K ◽  
Praneeth Gowd K ◽  
Ram Pavan S
Keyword(s):  
Antioxidant Activity ◽  
Radical Scavenging Activity ◽  
Iridoid Glycosides ◽  
Radical Scavenging ◽  
Anthelmintic Activity ◽  
Total Phenolic ◽  
Standard Drug ◽  
Flower Extract ◽  
Reducing Ability ◽  
Total Flavonoids Content

Nyctanthes arbour-tristis is a shrub belongs to the family Oleaceae. The flowers of this plant are fragrant since the presence of flavonol glycosides. It has also been reported for the presence of β-sitosterol, iridoid glycosides, tannins etc., and known to have immunostimulant, hepatoprotective, antiviral and antifungal activities. In the present study an attempt is made to identify antioxidant capacity and anthelminthic potential of methanolic flower extract of Nyctanthes arbour-tristis. Antioxidant activity was evaluated by total phenolic content assay, total flavonoids content assay, free radical scavenging activity and reducing ability methods. Anthelmintic activity was evaluated on Perithima posthuma using Piperazine citrate as standard drug. The results obtained for the above activities reveals that Nyctanthes arbour-tristis shows considerable antioxidant activity for all the methods and anthelminthic potential at 300 mg/ml. Keywords: arbour-tristis; antioxidant activity; anthelminthic activity; Perithima posthuma; Piperazine citrate.

Assessment of antioxidant and antibacterial activities of crude extracts of verbena officinalis Linn root or Atuch (Amharic)

2019 ◽  
Author(s):  
Chem Int
Keyword(s):  
Antioxidant Activity ◽  
Methanolic Extract ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Dry Weight ◽  
Antimicrobial Activities ◽  
Total Flavonoid ◽  
Root Extract ◽  
Antioxidant Power ◽  
Crude Extracts

Verbena officinalis Linn is a traditionally known medicinal plant which is used against a number of diseases including inflammatory conditions. In this study its antioxidant activity (reducing powers, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities), ferric reduction activity potential (FRAP), total flavonoid concentration and antimicrobial activities of 80%, 90%, 100% methanol and chloroform extracts of V. officinalis Linn root and 90% and100% methanol leaf extracts were determined. Its antioxidant activity increases with increase in amount of extract (10% to 40%v/v). Total flavonoid content (TFC) varied from 73.32±0.002 mgQE/100g of dry weight (90% methanol) to 42.39±0.032 mgQE/100g dry weight (chloroform), 2,2-diphenyl-1-picrylhydrazyl (DPPH), radical scavenging activity (%) was varied between 87.39% (90% methanol) to 45.57% (chloroform) while Ferric reducing antioxidant power was observed between 372.93±0.04 mgAAE/100 g extract (90% methanol) to 129.41±0.026 mgAAE/100 g chloroform in the root extract. The methanolic extract of the leaf showed less antioxidant activity than the methanolic extract of the root. Crude extracts of V. officinalis root showed various degree of antimicrobial activity towards drug resistance microbial pathogens. Growth inhibition tests against bacterial pathogens demonstrated concentration dependence. Moreover, gram positive bacteria were more susceptible to V. officinalis root extract when compared to gram negative bacteria. In general V. officinalis root and leave extracts possess strong antioxidant and antimicrobial activities.

PHYTOCHEMICAL CONSTITUENTS, ANTIOXIDANT ACTIVITY AND ACUTE ORAL TOXICITY OF POMEGRANATE (PUNICA GRANATUM L.) FRUIT PEEL EXTRACT

2019 ◽  
pp. 7-14
Author(s):  
Hai Trieu Ly ◽  
Tuan Anh Vo ◽  
Viet Hong Phong Nguyen ◽  
Thi My Sa Pham ◽  
Bich Thao Lam ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Gallic Acid ◽  
Radical Scavenging ◽  
Dry Weight ◽  
Punica Granatum ◽  
Acute Oral Toxicity ◽  
Oral Toxicity ◽  
Fruit Peel ◽  
Total Polyphenol ◽  
Punica Granatum L

Background: The natural antioxidants have an important role in the prevention of many diseases. The aim of study is to investigate phytochemical components, antioxidant activity and acute oral toxicity of Pomegranate (Punica granatum L.) fruit peel (PFP) extract. Materials and methods: Phytochemicals of PFP were determined by qualitative chemical tests, thin layer chromatography, total polyphenol and flavonoid contents. The PFP extract was evaluated for antioxidant activity by DPPH assay and MDA assay. In vivo acute oral toxicity test was conducted using Karber-Behrens method to determine LD50. Results: Results illustrated that PFP mainly contains flavonoids, alkaloids, tannins, triterpenes, saponins, and coumarins. PFP extract exhibited the total polyphenol and flavonoid contents with 189.97 mg gallic acid equivalent/g dry weight and 9.42 mg quercetin equivalent/g dry weight, respectively. The DPPH free radical scavenging and anti-lipid peroxidation activities of PFP extract were expressed with IC50 value of 4.80 μg/mL and 0.38 μg/ mL, sequentially. Simultaneously, the Dmax (the maximum dose administered to mice that no toxicity was observed) of PFP extract was determined to be 21.28 g/kg, equivalent to 35.64 g dried herb. Conclusion: The PFP extract is relatively safe and revealed high antioxidant activity. Key words: Punica granatum L.; polyphenols; flavonoids; gallic acid; quercetin; antioxidant activity; acute oral toxicity

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