NATTOKINASE

Degradative Effect of Nattokinase on Spike Protein of SARS-CoV-2

Takashi Tanikawa  1 , Yuka Kiba  2 , James Yu  3 , Kate Hsu  3 , Shinder Chen  3 , Ayako Ishii  4 , Takami Yokogawa  2 , Ryuichiro Suzuki  5 , Yutaka Inoue  1 , Masashi Kitamura  2

Abstract

The coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged as a pandemic and has inflicted enormous damage on the lives of the people and economy of many countries worldwide. However, therapeutic agents against SARS-CoV-2 remain unclear. SARS-CoV-2 has a spike protein (S protein), and cleavage of the S protein is essential for viral entry. Nattokinase is produced by Bacillus subtilis var. natto and is beneficial to human health. In this study, we examined the effect of nattokinase on the S protein of SARS-CoV-2. When cell lysates transfected with S protein were incubated with nattokinase, the S protein was degraded in a dose- and time-dependent manner. Immunofluorescence analysis showed that S protein on the cell surface was degraded when nattokinase was added to the culture medium. Thus, our findings suggest that nattokinase exhibits potential for the inhibition of SARS-CoV-2 infection via S protein degradation.

Citation

Tanikawa T, Kiba Y, Yu J, Hsu K, Chen S, Ishii A, Yokogawa T, Suzuki R, Inoue Y, Kitamura M. Degradative Effect of Nattokinase on Spike Protein of SARS-CoV-2. Molecules. 2022 Aug 24;27(17):5405. doi: 10.3390/molecules27175405. PMID: 36080170; PMCID: PMC9458005. https://pubmed.ncbi.nlm.nih.gov/36080170/

 

Nattokinase: A Promising Alternative in Prevention and Treatment of Cardiovascular Diseases

Chen  1 , Eileen M McGowan  2 , Nina Ren  3 , Sara Lal  2 , Najah Nassif  2 , Fatima Shad-Kaneez  2 , Xianqin Qu  2 , Yiguang Lin  2

Abstract

Cardiovascular disease (CVD) is the leading cause of death in the world and our approach to the control and management of CVD mortality is limited. Nattokinase (NK), the most active ingredient of natto, possesses a variety of favourable cardiovascular effects and the consumption of Natto has been linked to a reduction in CVD mortality. Recent research has demonstrated that NK has potent fibrinolytic activity, antihypertensive, anti-atherosclerotic, and lipid-lowering, antiplatelet, and neuroprotective effects. This review covers the major pharmacologic effects of NK with a focus on its clinical relevance to CVD. It outlines the advantages of NK and the outstanding issues pertaining to NK pharmacokinetics. Available evidence suggests that NK is a unique natural compound that possesses several key cardiovascular beneficial effects for patients with CVD and is therefore an ideal drug candidate for the prevention and treatment of CVD. Nattokinase is a promising alternative in the management of CVD.

Citation

Chen H, McGowan EM, Ren N, Lal S, Nassif N, Shad-Kaneez F, Qu X, Lin Y. Nattokinase: A Promising Alternative in Prevention and Treatment of Cardiovascular Diseases. Biomark Insights. 2018 Jul 5;13:1177271918785130. doi: 10.1177/1177271918785130. PMID: 30013308; PMCID: PMC6043915.

https://pubmed.ncbi.nlm.nih.gov/30013308/

 

Nattokinase: An Oral Antithrombotic Agent for the Prevention of Cardiovascular Disease

Yunqi Weng,1 Jian Yao,1 Sawyer Sparks,2 and Kevin Yueju Wang2,*

Abstract

Natto, a fermented soybean product, has been consumed as a traditional food in Japan for thousands of years. Nattokinase (NK), a potent blood-clot dissolving protein used for the treatment of cardiovascular diseases, is produced by the bacterium Bacillus subtilis during the fermentation of soybeans to produce Natto. NK has been extensively studied in Japan, Korea, and China. Recently, the fibrinolytic (anti-clotting) capacity of NK has been recognized by Western medicine. The National Science Foundation in the United States has investigated and evaluated the safety of NK. NK is currently undergoing a clinical trial study (Phase II) in the USA for atherothrombotic prevention. Multiple NK genes have been cloned, characterized, and produced in various expression system studies. Recombinant technology represents a promising approach for the production of NK with high purity for its use in antithrombotic applications. This review covers the history, benefit, safety, and production of NK. Opportunities for utilizing plant systems for the large-scale production of NK, or for the production of edible plants that can be used to provide oral delivery of NK without extraction and purification are also discussed.

Citation

Weng Y, Yao J, Sparks S, Wang KY. Nattokinase: An Oral Antithrombotic Agent for the Prevention of Cardiovascular Disease. Int J Mol Sci. 2017 Feb 28;18(3):523. doi: 10.3390/ijms18030523. PMID: 28264497; PMCID: PMC5372539. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5372539/

 

NATTOKINASE (Anti-blood clotting)

Nattokinase (NK) is a serine protease purified and extracted from natto, a traditional Japanese (cheese like) food produced from the fermentation of soybeans with the bacterium, Bacillus subtilis. [74-76] Recent studies demonstrated that a high natto intake was associated with decreased risk of total cardiovascular disease mortality and, in particular, a decreased risk of mortality from ischemic heart diseases. [77] Nattokinase has potent fibrinolytic, antithrombotic, and antiplatelet activity. [74;75;78-81] NK degrades fibrin directly and also increases the release of tPA with a subsequent increase in the formation of plasmin.

[82] Furthermore, NK enhances fibrinolysis through cleavage and inactivation of PAI-1. [76;81] In a study comparing the antiplatelet effects of NK and aspirin, NK was shown to display excellent antiplatelet aggregation and antithrombotic activities in vitro and in vivo, inhibiting thromboxane B2 formation from collagen-activated platelets. [83] In addition, in both animal and human studies, NK also has antihypertensive, anti-atherosclerotic, lipid-lowering, and neuroprotective actions. [75;81;84] Of particular relevance to patients with spike-related clotting, Nattokinase causes the proteolytic cleavage of both spike protein and amyloid proteins.

[85] In a randomized study, NK proved to be more effect than statins (simvastatin) in reducing carotid artery atherosclerosis. [86] Chen et al demonstrated that high dose NK (10 800 Fibrolytic Units [FU]/day; ~ 500 mg/day) reduced the thickness of the carotid artery intima-media and the size of the carotid plaque. [87] The authors reported a synergistic effect between NK and ASA.

Studies indicate that an oral administration of NK can be absorbed by the intestinal tract. [84;88] NK, unlike most proteins, is more resistant to the highly acidic gastric fluids in the stomach and can be absorbed in the later sections of the digestive tract. The optimal dose of nattokinase is unclear, however, a dose of 100-200 mg (4000- 8000 FU/day) twice daily has been suggested. While NK appears to have an excellent safety profile, [87;89] bleeding has rarely been reported in patients with risk factors for bleeding (advanced age, renal failure, hypertension, concomitant ASA, etc). [90;91] High concentrations of vitamin K2 in natto can reduce the INR when co-administered with warfarin; this may also occur with nattokinase supplements if vitamin K2 is not removed during the production process. Information regarding safety and efficacy in pregnancy and lactation is lacking.

Citations

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75. Weng Y, Yao J, Sparks S, Wang KY. Nattokinase: An oral antitrombotic agent for the prevention of cardiovascular disease. Int J Mol Sci 2017; 18:523. I-RECOVER: An approach to managing post-vaccine syndrome (1/9/2023) 41

76. Dabbagh F, Negahdaripour M, Berenjian A, Behfar A, Mohammadi F, Zamani M. Nattokinase: production and application. Applied Microbiology and Biotechnology 2014; 98:9199-9206.

77. Nagata C, Wada K, Tamura T, Konishi K, Goto Y, Koda S et al. Dietary soy and natto intake and cardiovascular disease mortality in Japanese adults: the Takayama study. Am J Clin Nutr 2017; 105:426-631.

78. Sumi H, Hamada H, Nakanishi K, Hiratani H. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematol 1990; 84:139-143.

79. Hsia CH, Shen MC, Lin JS, Wen YK, Hwang KL, Cham TM. Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. Nutrition Research 2009; 29:190-196.

80. Kurosawa Y, Nirengi S, Homma T, Esaki K, Ohta M. A single-dose of oral nattokinase potentiates thrombolysis and anti-coagulation profiles. Scientific Reports 2015; 5:11601.

81. Chen H, McGowan EM, Ren N, Lal S, Nassif N, Qu X et al. Nattokinase: A promising alternative in prevention and treatment of cardiovascular diseases. Biomarker Insights 2018; 13:1-8.

82. Yatagai C, Maruyama M, Kawahara T, Sumi H. Nattokinase-promoted tissue plasminogen activator release form human cells. Pathoyphysiol Haemost Thromb 2009; 36:227-232.

83. Jang JY, Kim TS, Cai J, Kim J, Kim Y, Shin K. Nattokinase improves blood flow by inhibiting platelet aggregation and thrombus formation. Lab Anim Res 2013; 29:221-225.

84. Fujita M, Ohnishi K, Takaoka S, Ogaswara K, Fukuyama R, Nakamuta H. Antihypertensive effects of continuous oral administraion of nattokinase and its fragment in spontaneously hypertensive rats. Biol Pharm Bull 2011; 34:1696-1701.

85. Tanikawa T, Kiba Y, Yu J, Hsu K, Chen S, Ishii A et al. Degradative effect of Nattokinase on spike protein of SARS-CoV-2. Molecules 2022; 27:5405.

86. Ren NN, Chen HJ, Li Y, Megowan GW, Lin YG. A clinical study on the effect of nattokinase on carotid artery atherosclerosis and hyperlipidemia [Chinese, Abstract in English]. Zhonghua Yi Vue Za Zhi 2017; 97:2038-2042.

87. Chen H, Chen J, Zhang F, Li Y, Wang R, Zheng Q. Effective management of atherosclerosis progress and hyperlipidemia with nattokinase: A clinical study with 1,1062 participants. Front Cardiovasc Med 2022; 9:964977.

88. Fujita M, Hong K, Ito Y, Misawa S, Takeuchi N, Kariya K et al. Transport of nattokinase across the rat intestinal tract. Biol Pharm Bull 1995; 18:1194-1196.

89. Gallelli G, Di Mizio G, Palleria C, Siniscalchi A, Rubino P. Data recorded in real life support the safety of Nattokinase in patients with vascular diseases. Nutrients 2021; 13:2031.

91. Ramachandran L, Aqeel A, Jafri A, Sidhu Y, Djirdeh TM. Nattokinase-associated hemoperitoneum in an elderly woman. Cureus 2022; 13:-e20074.
92. Chnag YY, Liu JS, Lai SL, Wu HS, Lan MY. Cerebellar hemorrhage provoked by combinaed use of nattokinase and aspirin in a patient with cerebral microbleeds. Inter Med 2008; 47:467-469.