Strain improvement, production and stability of Nattokinase from UV mutant strain of Pseudomonas aeruginosa CMSS

Abstract

Nattokinase (NK) is a potent fibrinolytic enzyme , which belongs to the second large family of serine proteases. NK gain significant attention in the treatment of hypertension and cardiovascular disorders (CVD's). Thus, a number of NK producers have been extensively studied, especially Pseudomonas sp. has extended the production level than Bacillus sp. with different properties. Thus the current study precedes with the strain improvement of NK producing UV mutated Pseudomonas aeruginosa CMSS by chemical mutagenesis. The potent mutant strain UV-EMS, treated with Ethyl Methyl Sulphonate (EMS) showed maximum NK activity. The maximum production of NK from mutant strain was determined at optimized parameters like pH 5 (1315.8 U mL-1), temperature at 37 o C (2413.3 U mL-1), shrimp shell as nitrogen source (2355.0 U mL-1) and sucrose as carbon source (3930.0 U mL-1).The activity of partially purified NK was stable at pH 7, temperature at 10 o C and 5mM of MgCl 2. The stability of NK was partially inhibited by SDS and completely inhibited by EDTA. The partially purified NK showed74% of in vitro clot lysis activity.

Comparative Studies on the Optimization of Staphylokinase and Nattokinase Produced From Staphylococcus Aureus and Bacillus Inaqusorum

“…to control. EDTA and silver nitrate showed almost same amount of decrease in the enzyme activity as shown in fig16 and fig17. The statistical analysis done by ANOVA showed that there is no significant difference between the inhibitors used in Staphylokinase and Nattokinase production as p value was greater than 0.05 as shown in table 13..…”

“…It is produced by fermentation by adding the bacterium Bacillus natto which is the preeminent nattokinase producer (Haritha, M. et al, 2011). It is a serine protease of the subtilisin family with 275 amino acid residues and molecular weight of 27,728 Dalton having potent fibrinolytic activity (Mohansrinivasan, V. et al, 2014). Nattokinase is in expensive and promote normal blood pressure, reduce whole blood viscosity and increase circulation being an effective supplement to support cardiovascular disease (Kim, W. et al, 1996).…”