MINI-REVIEW Nattokinase: production and application Fatemeh Dabbagh & Manica Negahdaripour & Aydin Berenjian & Abdolazim Behfar & Fatemeh Mohammadi & Mozhdeh Zamani & Cambyz Irajie & Younes Ghasemi Received: 24 August 2014 /Revised: 30 September 2014 /Accepted: 4 October 2014 /Published online: 22 October 2014 # Springer-Verlag Berlin Heidelberg 2014 Abstract Nattokinase (NK, also known as subtilisin NAT) (EC 3.4.21.62) is one of the most considerable extracellular enzymes produced by Bacillus subtilis natto. The main inter- est about this enzyme is due to its direct fibrinolytic activity. Being stable enough in the gastrointestinal tract makes this enzyme a useful agent for the oral thrombolytic therapy. Thus, NK is regarded as a valuable dietary supplement or nutraceu- tical. Proven safety and ease of mass production are other advantages of this enzyme. In addition to these valuable advantages, there are other applications attributed to NK in- cluding treatment of hypertension, Alzheimer’ s disease, and vitreoretinal disorders. This review tends to bring a brief description about this valuable enzyme and summarizes the various biotechnological approaches used in its production, recovery, and purification. Some of the most important applications of NK, as well as its future prospects, are also discussed. Keywords Bacillus subtilis . Nattokinase . Medical applications . Subtilisin NAT . Supplement . Fibrinolytic activity Introduction In 1987, Sumi et al. (1987) introduced a novel fibrinolytic enzyme from Natto (a typical and popular fermented soybean food in the Japanese diet) and named it nattokinase. Nattokinase (NK, also known as subtilisin NAT) (EC 3.4.21.62) is among the most considerable extracellular enzymes being produced by Bacillus subtilis natto (Hsieh et al. 2009; Liu et al. 2005; Unrean and Nguyen 2013). The present review aims to detail the current knowledge of the NK enzyme. This aim is followed by the critical review regarding its production, extraction, formu- lation, and applications. SCOPUS, Web of Science, and PubMed databases were reviewed until July 2014 using the “nattokinase” and “subtilisin NAT” keywords as the search terms. The lists of articles identified by this search strategy were studied and the relevant articles were selected. Nattokinase structure Natto is a traditional soybean food produced by B. subtilis natto fermentation. Natto is considered a rich source of valu- able products including NK and menaquinone-7 (Berenjian et al. 2014b). We previously demonstrated this strain as the dominant microorganism for the industrial production of these health-promoting compounds (Berenjian et al. 2012, 2013). NK is a serine protease composed of 275 amino acids (molecular weight 27.7 kDa and pI 8.6) which belongs to F. Dabbagh : M. Negahdaripour : F. Mohammadi : M. Zamani : Y. Ghasemi Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran F. Dabbagh : F. Mohammadi : M. Zamani : Y. Ghasemi (*) Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran e-mail: ghasemiy@sums.ac.ir A. Berenjian (*) School of Engineering, Faculty of Science and Engineering, The University of Waikato, Hamilton, New Zealand e-mail: aydin.berenjian@waikato.ac.nz A. Behfar Department of Food Science and Medical Hydrology, School of Pharmacy, Research Center for Safety Assessment, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran C. Irajie Department of Resource Development and Management, Shiraz University of Medical Sciences, Shiraz, Iran Appl Microbiol Biotechnol (2014) 98:9199–9206 DOI 10.1007/s00253-014-6135-3