Expression and purification of recombinant nattokinase in Spodoptera frugiperda cells

Xiaoxiang, Li; Wang, Xiaoli; Xiong, Shaoling; Cai, Lijun; Yu, Yang

doi:10.1007/s10529-007-9426-2

Cited by 14 publications

(9 citation statements)

References 11 publications

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“…There are many advantages, including its food origin and safety, to apply subtilisin NAT to thrombosis therapy. Recently, many genetic engineering methods, for example, heterologous expression by Escherichia coli or Spodoptera frugiperda cells, of producing subtilisin NAT have been reported (27,28). However, low expression efficiency of recombinant hosts, low yield, and instability of recombinant subtilisin NAT confine it to the application of industrial scale production.…”

Section: Discussionmentioning

confidence: 99%

A Simple and Cost-Saving Approach To Optimize the Production of Subtilisin NAT by Submerged Cultivation of Bacillus subtilis Natto

Tsai

Pan

2008

J. Agric. Food Chem.

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Subtilisin NAT, formerly designated nattokinase or subtilisin BSP, is a potent cardiovascular drug because of its strong fibrinolytic activity and safety. In this study, one Bacillus subtilis natto strain with high fibrinolytic activity was isolated. We further studied the optimal conditions for subtilisin NAT production by submerged cultivation and three variables/three levels of response surface methodology (RSM) using various inoculum densities, glucose concentrations, and defatted soybean concentrations as the three variables. According to the RSM analysis, while culturing by 2.93% defatted soybean, 1.75% glucose, and 4.00% inoculum density, we obtained an activity of 13.78 SU/mL. Processing the batch fermentation with this optimal condition, the activity reached 13.69 SU/mL, which is equal to 99.3% of the predicted value.

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Section: Discussionmentioning

confidence: 99%

A Simple and Cost-Saving Approach To Optimize the Production of Subtilisin NAT by Submerged Cultivation of Bacillus subtilis Natto

Tsai

Pan

2008

J. Agric. Food Chem.

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“…A modified Bac-to-Bac ® baculovirus expression system was used to express and produce active soluble NK in Spodoptera frugiperda insect cells [52]. The high production costs and longer duration needed for expression (~two weeks) do not support the use of insect cells as an NK production system.…”

Section: Recombinant Nattokinase Production Via Genetic Engineeringmentioning

confidence: 99%

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

Weng

Yao

Sparks

et al. 2017

IJMS

134

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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.

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“…NK gene from B. subtilis natto is also cloned and expressed in a variety of hosts including B. subtilis, Escherichia coli, Lactococcus lactis, and Spodoptera frugiperda insect cells (Chiang et al 2005;Li et al 2007;Liang et al 2007a, b;Liu and Song 2002;Nguyen et al 2013). Many efforts have been invested to increase the recombinant NK production.…”

Section: Nattokinase Production and Formulationmentioning

confidence: 99%

Nattokinase: production and application

Dabbagh

Negahdaripour

Berenjian

et al. 2014

Appl Microbiol Biotechnol

104

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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 interest 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 nutraceutical. 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 including 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.

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Expression and purification of recombinant nattokinase in Spodoptera frugiperda cells

Cited by 14 publications

References 11 publications

A Simple and Cost-Saving Approach To Optimize the Production of Subtilisin NAT by Submerged Cultivation of Bacillus subtilis Natto

A Simple and Cost-Saving Approach To Optimize the Production of Subtilisin NAT by Submerged Cultivation of Bacillus subtilis Natto

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

Nattokinase: production and application

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