Spectrophotometric bioassay method for urokinase.

INTRODUCTION: Urokinase is a potent plasminogen activator. Present Bio assay methods of Urokinase are very tedious. So a new simple spectrophotometric Bio assay method was developed to estimate thrombolytic activity of Urokinase. METHODS AND RESULTS: This Bio assay is designed for the quantitative in vitro spectrophotometric determination of Urokinase activity by utilizing its thrombolytic activity to carry out the lysis of plasma clots. The initial concentration of the plasma clots was adjusted to 0.2+/-0.01 absorbance and the linear decrease in the absorbance by addition of Urokinase concentration from 200 to 1200 IU/ml at lambda(max) 530 nm was studied. The activity of sample Urokinase can be quantified by comparing the absorbance of sample Urokinase with Urokinase standard Bio assay calibration curve and predicted in IU. The r(2) value of standard calibration curve was found out to be 0.993. The repeatability of the Bio assay was studied by performing the experiment six times with fresh plasma samples and the results showed significant similarity. DISCUSSION: We can conclude that the novel Bio assay method was found to be simple, economical, reproducible and accurate than the present Bio assay methods.

Media optimization studies for Serratiopeptidase production from Serratia marcescens ATCC 13880.

Production of an anti-inflammatory enzyme serratiopeptidase by fermentation with Serratia marcescens ATCC 13880 was studied to ascertain optimal nutritional conditions for large scale production. To study biosynthesis and production of serratiopeptidase by Serratia marcescens ATCC 13880, different physicochemical parameters were studied and optimized. The optimized medium contain, (g/l) glycerine 10.0, maltose 10.0 as carbon source, peptone 10.0 as organic nitrogen source, ammonium sulphate 10.0 as inorganic nitrogen source, dihydrogen phosphate 10.0, sodium bicarbonate 10.0, sodium acetate 10.0 as inorganic salt source, ascorbic acid 10.0 as stabilizer, distilled water 1000 ml and the optimized fermentation conditions were pH 7.0, temperature 37 degrees C and duration 24 hr. The modified fermentation medium produced 27.36 IU/ml of serratiopeptidase compared to 17.97 IU/ml in basal medium and the molecular weight of the purified serratiopeptidase was found to be 52 kD.