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The stability of Nattokinase during production

Samples collected in lab

The stability of nattokinase during production is always a question my clients ask after purchasing BSNK66 ® Nattokinase. Since stability is a major issue in manufacturing especially in the nutrient and food industry, the more we know about the specification of ingredients, the easier it will allow a better quality of final products and less waste of materials during production. Yet there are often so many consecutive factors that a slight change at one might impact the whole activity which we more often call it effect of the product. We will introduce you to the most suitable pH, temperature, and other catalysts that can help you get the best quality of nattokinase’ fibrinolytic activity.


The stability of nattokinase during production: Temperature

As the previous article (What is nattokinase enzyme used for?), we introduce that nattokinase’s main character of dissolving blood clots. Nattokinase has been shown that it has greater fibrinolytic activity than plasmin.

Long story short, as we and other essays indicate that the optimum temperature for nattokinase enzyme activity was approximately at 55℃. The nattokinase was relatively stable up to 55℃ but the stability dropped sharply at higher temperatures. It can incubate at 55℃ for 2 hours remained a relative activity of higher than 40% while its relative activity dropped down to less than 30% at 65℃ within 20 mins. In this article written by Hong-Ting Victor Lin, Guan-James Wu, and other 3 scientists you can check more about the decline of nattokinase activity in different temperatures (Purification and Characterization of Nattokinase from Cultural Filtrate of Red Alga Porphyra Dentata Fermented by Bacillus Subtilis N1).

So, this means that as long as the processing temperature is less than 55℃, the activity of nattokinase can be guaranteed. If the capsule is filled at room temperature, because the activity declines little, you can process it with confidence. In order to prevent the decline of nattokinase activity, generally, we Eiffel Biotech will study the processing process in detail with customers and give a certain more amount of activity to supplement it. For example, if our customer buys 20,000 FU/g of nattokinase, considering the attenuation of transportation and capsule filling, we will generally give away an activity of 1000-3000 FU/g more to ensure product quality.

The stability of nattokinase during production: Temperature

The stability of nattokinase during production: pH

The optimum pH for nattokinase activity is pH 8. But it remains over 90% activity at other tested pH values which indicates that nattokinase is active over a wide pH range. Since most of the production of nattokinase is making capsules, pills, and powder, we think that this pH might not be an important point when you manufacture nattokinase-related products.

One thing you need to be careful of is that the activity of nattokinase is completely lost when this enzyme was outside the pH range of 5 to 9.

The stability of nattokinase during production: pH


The stability of nattokinase during production: Inhibitors and Metal Ions

K+, Na+, Mg2+, Ca2+, Zn2+ ions show some minor inhibitory effect, which might be due to the fact that the substitutions or over loadings of metal ions K+, Na+, Mg2+, Ca2+, Zn2+ could change enzyme stability or activity. And Cu2+ (or SO42-) and Fe3+ ions at 10 mM enhanced the activity in a concentration-dependent manner, with being by 36.99% and 23.34%, respectively, at 10 mM.

What is the use of this information? Metal ions can reduce activity loss under high-temperature processing of nattokinase, but how to remove them in the subsequent steps is also a problem for processing plants.

This also provides us with an idea to make a product that combined Fn, Zn, Mg, Ca, and nattokinase.

The stability of nattokinase during production: Inhibitors and Metal Ions


Do you have any other questions? If you also want to process nattokinase, please contact us!



[1] Lin, Hong-Ting Victor, et al. “Purification and characterization of Nattokinase from cultural filtrate of red alga porphyra dentata fermented by Bacillus subtilis N1.” Journal of Marine Science and Technology 23.2 (2015): 240-248.

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