The Optimization of Protease Enzyme Extraction From Streblus Asper (Kesinai)

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

Download Full-text

Insights Studies for Certain Natural FDA Approved Polyphenolics and Repurposing for COVID-19

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11ispl1.3667 ◽

2020 ◽

Vol 11 (SPL1) ◽

pp. 1390-1395

Author(s):

Arafa Musa ◽

Ehab M. Mostafa ◽

Mohammad M. Al-Sanea ◽

Shaimaa R. Ahmed ◽

Gomaa Mostafa-Hedeab ◽

...

Keyword(s):

Docking Studies ◽

Human Populations ◽

Energy Score ◽

Protease Enzyme ◽

Human Disorders ◽

Variable Chemical ◽

Pancreatic Disorders ◽

New Treatment ◽

New Applications ◽

Death Cases

Natural polyphenolic drugs were approved for treatment of various diseases. Diosmin, rutin, quercetin, aesculin, genistein, hesperidin and silybin are known for their safety and have been applied for several human disorders including cancer, cardiovascular disorders, atherosclerosis, oxidative stress, capillary fragility, liver and pancreatic disorders and others. As the structures of the selected polyphenolic compounds possess variable chemical moieties with diverse chemical and electronic characters, these properties have been employed for further insights studies to predict new applications concerning the newly occurred pandemic. COVID-19 is a terrible disease that attacked millions of human populations at the end of year 2019. As the number of death cases has increased to exceed one million of humans, the early discovery of new treatment from previously approved and safe drugs is the main target of this study. Employing the putative docking studies for the selected polyphenolic drugs were done and the results were promising, all tested drugs exhibited high affinity with the selected five proteins of protease enzyme, the docking score ranged from -8.4461 to -26.1691 kcal/mol with 3-7 variable interaction bonds. Among the tested drugs, diosmin appeared as the most promising drug as it exhibited the highest energy score and interaction bonds with the most of proteins.

Download Full-text

The Influence of Hydrogen atoms on the Performance of Radial Distribution Function Based Descriptors in the Chemoinformatic Studies of HIV-1 Prote-ase Complexes with Inhibitors.

Current Drug Discovery Technologies ◽

10.2174/1570163817666200102130415 ◽

2020 ◽

Vol 17 ◽

Author(s):

Jurica Novak ◽

Maria A. Grishina ◽

Vladimir A. Potemkin

Keyword(s):

Distribution Function ◽

Radial Distribution Function ◽

Radial Distribution ◽

Current Model ◽

Direct Interaction ◽

Hydroxyl Group ◽

Linear Regression Method ◽

Hydrogen Atoms ◽

Protease Enzyme ◽

Hiv 1

: In this letter the newly introduced approach based on the radial distribution function (RDF) weighted by the number of va-lence shell electrons is applied for a series of HIV-1 protease enzyme and its complexes with inhibitors to evaluate the influ-ence of hydrogen atoms on the performance of the model. The multiple linear regression method was used for the selection of the relevant descriptors. Two groups of residues having dominant contribution to the RDF descriptor are identified as relevant for the inhibition. In the first group are residues like Arg8, Asp25, Thr26, Gly27 and Asp29, which establish direct interaction with the inhibitor, while the second group consists of the amino acids at the interface of the two homodimer sub-units or with the solvent. The crucial motif pointed out by our approach as the most important for inhibition of the enzyme’s activity and present in all inhibitors is hydroxyl group that establish hydrogen bond with Asp25 side chain. Additionally, the comparison to the model without hydrogen showed that both models are of similar quality, but the downside of the current model is the need for the determination of residues’ protonation states.

Download Full-text

Constraints in Adoption ofModern Farm Machines by Tribal Farmers in Ramgarh District of Jharkhand

Journal of AgriSearch ◽

10.21921/jas.v6i03.16216 ◽

2019 ◽

Vol 6 (03) ◽

Author(s):

PK SUNDARAM ◽

BIKASH SARKAR ◽

UJJWAL KUMAR ◽

AP ANURAG ◽

DK RAGHAV ◽

...

Keyword(s):

Cell Line ◽

Normal Cell ◽

Human Cancer ◽

Skim Milk ◽

Exchange Chromatography ◽

Protease Production ◽

Normal Cell Line ◽

Protease Enzyme ◽

Tribal Farmers ◽

Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) andamp; the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method andamp; it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column andamp; the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , andamp; one normal cell line Ref ( Rat embryonic fibroblast ). The cancer andamp; normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8andamp; 0.16 mg/ml) then incubated for additional 48h at 37C 0 andamp; the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andamp;slight toxicity ( 37.12% )on normal cell line (Ref) in a concentration (0.8mg/ml).

Download Full-text

Convergent Chemical Synthesis and Crystal Structure of a 203 Amino Acid “Covalent Dimer” HIV-1 Protease Enzyme Molecule

Angewandte Chemie International Edition ◽

10.1002/anie.200604087 ◽

2007 ◽

Vol 46 (10) ◽

pp. 1667-1670 ◽

Cited By ~ 139

Author(s):

Vladimir Yu. Torbeev ◽

Stephen B. H. Kent

Keyword(s):

Crystal Structure ◽

Amino Acid ◽

Chemical Synthesis ◽

Enzyme Molecule ◽

Synthesis And Crystal Structure ◽

Protease Enzyme ◽

Covalent Dimer ◽

Hiv 1

Download Full-text

Streblus asper Lour. exerts MAPK and SKN-1 mediated anti-aging, anti-photoaging activities and imparts neuroprotection by ameliorating Aβ in Caenorhabditis elegans

Nutrition and Healthy Aging ◽

10.3233/nha-210121 ◽

2021 ◽

pp. 1-17

Author(s):

Mani Iyer Prasanth ◽

James Michael Brimson ◽

Dicson Sheeja Malar ◽

Anchalee Prasansuklab ◽

Tewin Tencomnao

Keyword(s):

Oxidative Stress ◽

Caenorhabditis Elegans ◽

Stress Resistance ◽

Vital Role ◽

Transgenic Strain ◽

Wild Type ◽

Neuroprotective Effects ◽

C Elegans ◽

Streblus Asper

BACKGROUND: Streblus asper Lour., has been reported to have anti-aging and neuroprotective efficacies in vitro. OBJECTIVE: To analyze the anti-aging, anti-photoaging and neuroprotective efficacies of S. asper in Caenorhabditis elegans. METHODS: C. elegans (wild type and gene specific mutants) were treated with S. asper extract and analyzed for lifespan and other health benefits through physiological assays, fluorescence microscopy, qPCR and Western blot. RESULTS: The plant extract was found to increase the lifespan, reduce the accumulation of lipofuscin and modulate the expression of candidate genes. It could extend the lifespan of both daf-16 and daf-2 mutants whereas the pmk-1 mutant showed no effect. The activation of skn-1 was observed in skn-1::GFP transgenic strain and in qPCR expression. Further, the extract can extend the lifespan of UV-A exposed nematodes along with reducing ROS levels. Additionally, the extract also extends lifespan and reduces paralysis in Aβ transgenic strain, apart from reducing Aβ expression. CONCLUSIONS: S. asper was able to extend the lifespan and healthspan of C. elegans which was independent of DAF-16 pathway but dependent on SKN-1 and MAPK which could play a vital role in eliciting the anti-aging, anti-photoaging and neuroprotective effects, as the extract could impart oxidative stress resistance and neuroprotection.

Download Full-text

Biological activities of spores and metabolites of some fungal isolates on certain aspects of the spiny bollworms Earias insulana (Boisd.) (Lepidoptera: Noctuidae)

Egyptian Journal of Biological Pest Control ◽

10.1186/s41938-019-0192-y ◽

2019 ◽

Vol 29 (1) ◽

Cited By ~ 1

Author(s):

Eman Mohammed Abd-ElAzeem ◽

Warda Ahmed Zaki El-Medany ◽

Hend Mohammed Sabry

Keyword(s):

Biological Activities ◽

Larval Mortality ◽

Adult Emergence ◽

Pupal Weight ◽

Earias Insulana ◽

Dead Larva ◽

Fungal Isolates ◽

Protease Enzyme ◽

Newly Hatched Larvae ◽

Lepidoptera Noctuidae

AbstractBiological activities of spores and metabolites of some fungi isolated from dead larva of the spiny bollworms (SBW), Earias insulana (Boisd.) (Lepidoptera: Noctuidae), against the newly hatched larvae of the pest were carried out. Results showed that the fungi Metarhizium anisopliae, Acremonium sp., and Paecilomyces variotii had affected the newly hatched larvae of (SBW). Acremonium sp. was the most potent one as it had the highest newly hatched larval mortality percentage (65 and 58.33%) for its spore suspension and metabolites, respectively, while the lowest one (41%) was for P. variotii metabolites. Also, spore suspensions of the all fungal isolates had the highest larval mortality than fungal metabolites. Studying the enzymatic activity showed that Acremonium sp. produced protease enzyme on media containing gelatin, which caused the highest larval mortality (72.22%).These isolates showed different effects on all stages of the pest and decreased pupal weight, adult emergence percentages, deposited eggs, and hatchability percentages than the control. Identification of Acremonium sp. EZ1 was confirmed using 18 s rRNA and its accession number MN25101.

Download Full-text

Determination of Clenbuterol, Salbutamol, and Cimaterol in Bovine Retina by Electrospray Ionization–Liquid Chromatography–Tandem Mass Spectrometry

Journal of AOAC International ◽

10.1093/jaoac/87.1.31 ◽

2004 ◽

Vol 87 (1) ◽

pp. 31-38 ◽

Cited By ~ 18

Author(s):

Joseph H W Lau ◽

Cheang S Khoo ◽

John E Murby

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Tandem Mass Spectrometry ◽

Electrospray Ionization ◽

Tandem Mass ◽

Bovine Retina ◽

Chromatography Tandem Mass Spectrometry ◽

Protease Enzyme ◽

Liquid Chromatography Tandem Mass

Abstract An electrospray ionization–liquid chromatography–tandem mass spectrometry (ESI/LC/MS/MS) method was developed for the simultaneous determination of the β-agonists clenbuterol, salbutamol, and cimaterol in bovine retina. The tissue was homogenized in alkaline buffer and spiked to give 10, 15, and 20 ng/g each of the 3 analytes together with the internal standards d6-salbutamol and d6-clenbuterol. The mixture was incubated with protease enzyme to release any protein-bound analytes and then made alkaline before extraction with isobutanol. The extract was dissolved in water and transferred to a clenbuterol immunoaffinity column. After washing, the analytes were eluted and analyzed by ESI/LC/MS/MS using a C18 column with acetic acid–methanol as mobile phase. No interferences were observed from the spiked retina extract at the various single-reaction monitoring modes. Average recoveries for clenbuterol, salbutamol, and cimaterol were 94, 85, and 87% with coefficients of variation (CVs) of 9.4, 9.9, and 8.6%, respectively. A correlation coefficient of r2 = 0.9999 was obtained for all analytes. The limits of detection for clenbuterol, salbutamol, and cimaterol, determined from 3 times the standard deviation of 7 replicates of the lowest spike, were 2.5, 3.5, and 2.0 ng/g with CVs of 8.9, 11.6, and 7.2%, respectively.

Download Full-text