19-08-2014, 11:30 AM
The present work was undertaken with the following objectives: (1) To study fermentative production of nattokinase using Bacillus natto. The media optimization using one factor-at-a- time and statistical method viz. Response Surface Methodology (RSM) (2) To study downstream processing of nattokinase. (3) Characterization of nattokinase.
Nattokinase, a fermentation product has attracted worldwide attention because of its health benefits and long history in Japanese food. Nattokinase (E.C. 3.4.21.62) is a novel fibrinolytic enzyme and can be considered to be a promising agent for thrombosis therapy. The fibrinolytic enzymes were successively discovered from different microorganisms, the most important among which is the genus Bacillus from traditional fermented foods. The human body produces several types of enzymes for making thrombus, but only one main enzyme for breaking it down and dissolving it is plasmin. The properties of nattokinase closely resemble that of plasmin which makes nattokinase a particularly potent thrombolytic agent.
The present work was undertaken with the following objectives: (1) To study fermentative production of nattokinase using Bacillus natto. The media optimization using one factor-at-a- time and statistical method viz. Response Surface Methodology (RSM) (2) To study downstream processing of nattokinase. (3) Characterization of nattokinase.
In first step, optimization of nattokinase production was carried out for various parameters like pH, inoculum size, carbon source, nitrogen source and salt ions. The optimization was carried out by one factor at-a-time followed by response surface methodology (RSM).
One factor at-a-time method resulted in increase of nattokinase production from 4.2 FU/gm to 11.67 FU/gm. Further increase in nattokinase production from 11.67 FU/gm to 13.55 FU/gm was obtained using RSM.
Purification of nattokinase was done by ammonium sulphate precipitation followed by gel permeation chromatography (GPC). Characterization of enzyme was carried out to study the enzyme activity with respect to variations in pH and temperature.