Some properties of the extracellular, cell bound and intracellular protease of mucor hiemalis

This study was undertaken to provide some basic aspects of the extracellular, intracellular and cell-bound protease of M. Hiemalis. was also tested for the production of milk-clotting enzymes. It was found that the protease whether obtained extracellularly or intracellulary exhibited milk-clotting action, although it was very weak and took about 8 hours to clot the milk. Intracellular protease showed maximum activity when it was extracted with 7.8 pH phosphate buffer. It was most active when incubated at 35°C. The protease hydrolysed casein and the rate of hydrolysis increased with the increase in time. Similar results were obtained with the extracellular and cell-bound protease

Separation and partial characterization of two of the extracellular endoglucanases of cellulomonas flavigena

Two extracellular endoglucanases of Cellulomonas flavigena were purified by acetone precipitation, gel filtration, ion-exchange chromatography and preparative gradient PAGE and named as endoglucanases E1 and E2. Both of these were only active against carboxymethyl cellulose [CMC] and their Km values were determined to be 7.8 and 20 g/l, respectively. Each of the enzyme gave glucose along with cellobiose as end product. Both the endoglucanases were most active at pH 6.5 - 7.0 and at 45 - 50°C. Each of the enzyme was inhibited by 20 mM glucose and cellobiose. Heavy metal ion e.g. Ag+ and Fe++ and B-mercaptoethanol strongly inhibited both the enzymes whereas slight inhibition was observed with Ca++. The enzymes are stable in the pH range of 3 - 10 and below 30°C. Molecular weights as determined by SDS-PAGE are 75,000 for endoglucanase E1 and 60,000 for the endoglucanase E2

Lipid class and percentage of cotyledons and primary roots in zea mays [neelum]

An investigation was conducted to study the variation in the lipid class and fatty acid composition of the individual lipid fraction in Zea mays [Neelum] cotyledons and primary roots during germination to various root length. There was loss in both the dry weight and lipid content in the cotyledons, while in the roots the dry weight increased and decreased. During germination the relative amount of the neutral lipids were utilized at a faster rate than polar lipid in both the cotyledons and primary roots indicating that the seeds utilized the fat reserves to germinate. The resting seeds contained lesser amount of polar lipids as glycolipids and phosphatidyi ethanolamine and its lyso compound were absent. In the lipid class composition of the primary root significant changes indicated the function of neutral glycerides as a source of reserve energy while the glycolipids and phosphoglycerides played a role as component of the membrane systems in the growing root

Purification and partial characterization of extracellular lipases of mucor hiemalis

Extracellular lipases of Mucor hiemalis were purified from the culture grown in shake flasks for 96 hours. The broth was saturated with ammonium sulfate to 70%, the precipitates were dissolved and the solution desalted through a Sephadex G-25 column. The crude preparation was fractionated by chromatography with DEAE-Sephadex A-50 using a linear salt gradient. The two enzyme activities as obtained by pooling active fractions were purified further by passing each of these through a Sephadex G-l50 column. Each of the enzymes, F 1 and F 2, was found to be homogeneous by SDSPAGE. Enzyme actitives were also detected on polyacramide gel rods incorporated with 0.1% olive oil as substrate. Molecular weights of lipases F 1 and F 2 as determined by gel filteration with Sephadex G-150 were found to be 45, 710 and 19, 950, while their pH optima was 8.9 and 7.6, respectively. Lipase F 1 was purified 47.85 times with specific activity of 6.7 U/mg while lipase F2 showed 42.5 times purification with specific activity of 5.95 U/mg protein

Preparation and biological evaluation of Tc99m-Sn phosphate colloid for the study of reticuloendothelial system

A colloid of sodium dihydrogen phosphate and stannous chloride was prepared at pH 6.0 which could be instantly labellea with Tc [99m]. Labelling yield was over 96% as determined by paper chromatography, ultrafiltration and gel chromatography. Effect of heat, pH, electolytes and concentration on the stability of the colloid was studied by visual inspection. Particle size of the colloid was adjusted by observing agglomeration and comparing with other colloids like Tc [99m] Tin and Tc [99m] Sulphur. Biodistribution study performed in mice showed 70% of the colloid uptake in liver, 4% in spleen and 7% in bone marrow at one hour post-injection of Tc [99m]-Sn-PO4 colloid. Time-activity curve drawn showed maximum uptake in the liver area at 15 minutes post-injection. The radiocolloid showed a potential in the study of liver malfunctions in humans. No lung uptake was detected and no sign of toxicity was observed