Protease Production by Different Thermophilic Fungi

Submerged cultures of 10 thermophilic fungi, grown at 45° in a medium containing 4% casein, 0.4% glucose, 0.3% yeast extract and salts, were surveyed for extra- and intra-cellular protease production. The yield, thermostability, pH optimum, and inhibitor sensitivity of the crude proteases were determined in each case. The inhibitors used were diisopropylfluorophosphate (DFP), ethylenediaminetetraacetic acid (EDTA), and p-chloromercuribenzoate (pCMB).Seven of the thermophiles produced a substantial quantity of protease, particularly extracellular protease. In comparison to mesophilic fungi most proteases appeared to be unusually thermostable. Neutral and alkaline proteases were most common, as were proteases inhibited by DFP and pCMB. Finally in most of the cases a sensitivity to more than one inhibitor indicated the existence of more than one type of protease.

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Cytotoxic Effect of Pseudomonas aurogenosa Protease on Cancer Cells

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.10.3.7 ◽

2019 ◽

Vol 10 (03) ◽

Author(s):

Ghanyia J. Shanyoor ◽

Fatima R. Abdul ◽

Nehad A. Taher ◽

Ihsan A. Raheem

Keyword(s):

Cell Line ◽

Normal Cell ◽

Cytotoxic Effect ◽

Human Cancer ◽

Skim Milk ◽

Exchange Chromatography ◽

Protease Production ◽

Normal Cell Line ◽

Protease Enzyme ◽

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 ) 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).

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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).

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