Production and Purification of Nattokinase
Nattokinase is extract from fermented soybean product–natto, a traditional Japanese food which using bacterium ( Bacillus Subtilis ) fermentation of soybeans. And Dr Hiroyuki Sumi discovered the artificial fibrin can be dissolved by the natto in Chicago University Medical School. Then he and his team extracted an kind of enzyme from natto which not only can dissolve fibrin but also a plasmin substrate. He named this enzyme ” nattokinase”
Nattokinase is an enzyme mainly get from the natto, the fermanted soybean by Bacillus subtilis. Other fermented soybean -based foods like Chinese douchi, kidney beans, Thai thua nao and etc by Bacillus subtilis can produce a similar enzyme also called nattokinase.
The traditional way to make natto is very simple and easy which can make at home straightforward. The key and important to matto natto is Bacillus subtilis. The Bacillus subtilis fermented natto can keep activity at PH of 6-12 and resist the temperatures up to 60 degree. Its viability and metabolic activity of NK products can maintain more than 6 months. Boiled the soybean with Bacillus subtilis n the room temperature to ferment and wait more than 24 hours until its surface covered with sticky substanced which bacterium produce and composed of glutamate polymers. Different from the homemade, quantity production of NK to majorization the condition of fermenttation by productive practive to maximization producing yield of the NK by Bacillus subtilis, including the condition of right temperature, PH value, and fermentation time. In order to increase the Nattokinase yield, lots of nutriton ability have been examined to use, like yeast extract, shrimp shell, glycerol, and soy peptone. Compared with the yield obtained by batch fermentation, the optimal feeding strategy used in the fed-batch fermentation method significantly improved the yield of Nattokinase.
Different from the simple fermantation natto, the extract ,Production and Purification of Nattokinase is very difficult. Several processing include organic solvent, salting out proteins, Several steps are required, including homogenization with an organic solvent, salting out the proteins, protein ion-exchange chromatography and dialysis, etc. And we in factory, our researchers and enginners already get a high -repid and efficient techniques to extract and purify the NK. Thorat and Garg find and practice a high purify NK technique –three phase partitioning (TPP) by combining t-butanol (1.5× to crude extract) and ammonium sulphate (30% W/V) to precipitate the NK protein. Its results required the PH 8.0 and temperature 37 degree for guarantee the NK activity varies.
In Japan, the natto mainly used by a common food, but its mild odor and stringy texture is difficulty to accept by most of person. Other countries use NK mainy taking by capsules which made of the vegetable- based material as the therapeutic medicine for thrombosis. NK products are currently in the spotlight at the federal drug administration (FDA) because of the high purity requirements for single constituent entities. Therefore, recombination techniques have been explored to increase the amount and purity of NK produced.More Scientific research and development of production and purification of Nattokinase, please check here
