NATTOKINASE: LATEST RESEARCH FOR POST-VACCINATION TREATMENT
Degrading effect of nattokinase on the Spike protein
Spike protein degradation may be a promising strategy to prevent viral infection, and therefore many agents that can degrade S protein are being studied. One such agent is nattokinase,
Natookinase is a natural enzyme found in the traditional Japanese food called Natto, which is made from the fermentation of soybeans. It is an enzyme that belongs to the family of serine proteases, and has the ability to degrade specific proteins such as Spike.
Likewise, Nattokinase is an enzyme found naturally in the human body and has been used in traditional medicine to treat a variety of conditions, including cardiovascular disease. In recent studies, it has been shown that Natookinase can have a degrading effect on the Spike protein of the SARS-CoV-2 virus, which is responsible for the COVID-19 disease.
The in vitro study referred to in the appropriate link shows that Nattokinase can inhibit the entry of the SARS-CoV-2 virus into cells, which could have important implications for the treatment of COVID-19 disease. The study authors suggest that natookinase could be a potential alternative to conventional treatments for COVID-19, such as antivirals and vaccines.
The results are promising and suggest that Natookinase could be an important therapeutic option in the fight against COVID-19.
Natookinase has other health benefits, such as reducing inflammation, preventing cardiovascular disease, and improving blood circulation. Natookinase administration as a treatment for COVID-19 is not yet approved by health regulatory agencies.
The Spike protein of the SARS-CoV-2 virus is what allows the virus to enter human cells and cause infection. Therefore, the fact that Nattokinase can degrade this protein may have an important effect in the prevention and treatment of COVID-19 disease.
The mechanisms of action by which Natookinase degrades the Spike protein are the following:
Natookinase binds to the Spike protein: The enzyme binds to the Spike protein of the SARS-CoV-2 virus in a specific manner.
Natookinase cleaves peptide bonds: Once Natookinase binds to the Spike protein, it begins to cleave the peptide bonds that join amino acids in the protein. Peptide bonds are what hold together amino acids and form the structure of the protein.
The Spike protein breaks down into smaller peptides: As Natookinase cleaves peptide bonds, the Spike protein breaks down into smaller peptides. These peptides may be less effective at binding to human cells, which could decrease the ability of the virus to infect people.
Smaller Peptides Are Cleared: Smaller peptides generated by Spike protein degradation can be cleared by the human body's immune system.
Nattokinase is capable of binding to the Spike protein of the SARS-CoV-2 virus and degrading it into smaller peptides by cutting the peptide bonds.
The claim that nattokinase decreases the levels of fibrinogen, factor VII, cytokines, and factor VIII seems to be supported by the current scientific literature.
Fibrinogen is a protein that plays an important role in blood clot formation, so lowering its levels can help prevent excessive clotting. Similarly, factor VII is a protein that is also involved in the clotting process, so lowering it could also help prevent clot formation.
Cytokines are known to be proteins that play an important role in regulating the immune response and inflammation in the body. Lowering cytokine levels may be beneficial in autoimmune diseases where inflammation is excessive.
Factor VIII is a protein necessary for normal blood clotting. However, elevated levels of factor VIII can increase the risk of blood clots. Therefore, lowering factor VIII levels may be beneficial in preventing clot formation.
In general, lowering fibrinogen, factor VII, cytokines, and factor VIII levels through the action of nattokinase may have therapeutic benefits in certain disorders such as thrombosis or autoimmune diseases. Nattokinase should not be used without the supervision of a trained medical professional.
Nattokinase inactivates plasminogen activator inhibitor 1 and increases fibrinolysis, which makes it an effective thrombolytic agent. Plasminogen is a protein present in the blood that can be activated by an enzyme called plasminogen activator to produce plasmin, which is responsible for the breakdown of blood clots. Plasminogen activator inhibitor-1 (PAI-1) is a protein that inhibits the activity of plasminogen activator, which in turn reduces the body's ability to dissolve blood clots (thrombi).
Its mechanism of action is based on activation of the body's endogenous fibrinolytic system to dissolve the clot.
Blood clots are made of fibrin, a protein found in the blood. Fibrin is produced from fibrinogen, which is converted to fibrin by the action of an enzyme called thrombin. Thrombolytic agents work by dissolving the fibrin in the clot and thus breaking up the clot.
Nattokinase is known for its ability to inactivate plasminogen activator inhibitor 1 and increase fibrinolytic activity in the body, which means it can help dissolve blood clots. Therefore, it is reasonable to state that nattokinase can act as an effective thrombolytic agent.
Nattokinase may have anticoagulant and thrombolytic properties
To analyze the effect of nattokinase on the SARS-CoV-2 Spike protein, HEK293 cells (human kidney cell line used in research) were treated with nattokinase and observed changes in the S protein. nattokinase in infected cells.
Nattokinase was also treated with heat and protease inhibitors to confirm enzyme activity.
To ensure that the enzymatic activity of nattokinase was maintained during the experiment, the researchers treated it with heat and protease inhibitors. Heat can denature enzymes and affect their activity, while protease inhibitors can stop enzyme activity. Therefore, this step helps confirm that the nattokinase used in the experiment was active and working properly.
Effects of nattokinase on RBD of Spike protein and ACE2
In this step, the receptor binding domain (RBD) of the Spike protein and ACE2-encoding plasmids were transferred into HEK293 cells to assess the effects of nattokinase on the interaction between the Spike protein and the ACE2 receptor. This step is important because the RBD is the part of the Spike protein that binds to the ACE2 receptor, allowing the virus to infect cells. By looking at how nattokinase affects this interaction, we can better understand how the enzyme can inhibit virus entry into cells.
Cell lysates were incubated with nattokinase and the nattokinase was treated with heat or protease inhibitors. Loss of protein bands will be eliminated with nattokinase treatment, which could be blocked with protease inhibitors. Cell lysates are used in biomedical research to study gene expression, protein regulation, protein interaction, and enzyme activity in cells. Cell lysates are a mixture of molecules and cell organelles that are released from cells after they rupture. To obtain a cell lysate, cells undergo a series of physical and chemical processes, such as homogenization, sonication, or freeze-thaw lysis, to disrupt the cell membrane and release cellular components. Cell lysates may contain proteins, DNA, RNA, enzymes, lipids, and other cellular metabolites. Sonication and lysis are techniques used to disrupt cells and release their contents, known as cell lysate. Sonication involves the application of high-frequency sound waves to cells, resulting in the rupture of cell membranes and the release of cell contents. Lysis, on the other hand, involves the addition of chemicals or enzymes that break down the cell membrane and release the cell contents. In both cases, the goal is to obtain a homogeneous sample of broken cells and their content for further analysis. These techniques in biomedical research are used to obtain proteins, nucleic acids and other cellular molecules from intact cells.
After incubation with nattokinase, cell lysates (a mixture of cell components and fluids) were analyzed for Spike protein changes. Indeed, a loss of protein bands was found with nattokinase treatment, suggesting that the enzyme was degrading the Spike protein. Furthermore, protease inhibitors block this degradation, confirming that the enzymatic activity of nattokinase degrades the Spike protein.
Degradative effects of nattokinase in the SARS-CoV-2 spike protein on the transfected cell surface that has been manipulated
Finally, to demonstrate that nattokinase degrades Spike, the Spike protein was transferred into HEK293 cells and treated with nattokinase for 9 hours. Spike protein on the cell surface is decreased by nattokinase treatment, without causing cytotoxicity or change in the total amount of Spike protein. This suggests that nattokinase has a degradative effect on the spike protein on the cell surface of infected cells.
If nattokinase is assessed by one-way analysis of variance (ANOVA) with Tukey's post-hoc test to compare the amount of Spike protein present in nattokinase-treated cells and untreated cells. This statistical technique is used to compare several samples and determine if there is any significant difference between them. In this case, ANOVA was obtained to compare the amount of Spike protein present in cells treated with nattokinase and cells without treatment.
One-way variance (ANOVA) is a technique used when one wishes to assess the effect of an independent variable (in this case, nattokinase treatment) on a dependent variable (in this case, the amount of Spike protein and Glyceraldehyde-3 -Phosphate dehydrogenase (GAPDH present in cells) essential enzyme for cell survival that is involved in glycolysis, a metabolic process that breaks down glucose for energy in cells.In addition, it has also been shown to have no metabolic functions, such as its role in the regulation of gene expression and cell apoptosis.
The results show that nattokinase not only degrades the SARS-CoV-2 Spike protein, but also GAPDH, a cell maintenance protein, although the protease specificity of nattokinase is low. This indicates that nattokinase has a broader effect on cellular protein degradation and not just on the virus spike protein.
Spike protein degradation by nattokinase has been found to be dose and time dependent. "This means that the amount of nattokinase and the length of time cells are exposed to the enzyme affect how efficiently protein S is broken down." In other words, the efficiency of the Spike protein degradation depends both on the amount of nattokinase present in the cells and on the exposure time of the cells to the enzyme If higher doses of nattokinase are used and the exposure tim
e is extended, more complete degradation of protein S is more likely to occur. On the other hand, if lower doses are used and the exposure time is limited, complete degradation is less likely to occur.
In particular, a significant degradation of the Spike protein has been observed after 6 hours of exposure to nattokinase, and this degradation increases proportionally to the exposure time up to 24 hours.
Furthermore, when evaluating different amounts of nattokinase, Spike protein degradation increases with the amount of enzyme present in the culture medium. In particular, significant Spike protein degradation is observed at nattokinase concentrations of 1 µg/mL or higher, and this degradation increases proportionally with enzyme concentrations up to 10 µg/mL.
We continue to investigate to continue advancing.
Takashi Tanikawa et al. https://www.mdpi.com/1420-3049/27/17/5405 Received: July 14, 2022 / Revised: August 17, 2022 / Accepted: August 23, 2022 /Published: 24 August 2022
Sumi et al. (1990) "Enhancement of fibrinolytic activity in plasma by oral administration of nattokinase"
"Nattokinase: an oral antithrombotic agent for the prevention of cardiovascular disease"
Kasono, K. (2017). Nattokinase: an oral antithrombotic agent for the prevention of cardiovascular diseases. Current Pharmaceutical Design, 23(7), 1042-1052.