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Strain Screening, Fermentation, Separation, and Encapsulation for Production of Nattokinase Functional Food

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Abstract

This study presents a novel and integrated preparation technology for nattokinase functional food, including strain screening, fermentation, separation, and encapsulation. To rapidly screen a nattokinase-productive strain, PCR-based screening method was combined with fibrinolytic activity-based method, and a high productive strain, Bacillus subtilis LSSE-22, was isolated from Chinese soybean paste. Reduction of poly-γ-glutamic acid (γ-PGA) concentration may contribute to separation of nattokinase and reduction of late-onset anaphylaxis risk. Chickpeas were confirmed as the favorable substrate for enhancement of nattokinase production and reduction of γ-PGA yield. Using cracked chickpeas, the nattokinase activity reached 356.25 ± 17.18 FU/g (dry weight), which is much higher than previous reports. To further reduce γ-PGA concentration, ethanol fractional extraction and precipitation were applied for separation of nattokinase. By extraction with 50 % and precipitation with 75 % ethanol solution, 4,000.58 ± 192.98 FU/g of nattokinase powders were obtained, and the activity recovery reached 89 ± 1 %, while γ-PGA recovery was reduced to 21 ± 2 %. To improve the nattokinase stability at acidic pH condition, the nattokinase powders were encapsulated, and then coated with methacrylic acid–ethyl acrylate copolymer. After encapsulation, the nattokinase was protected from being denatured under various acid conditions, and pH-responsible controlled release at simulated intestinal fluid was realized.

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Acknowledgments

This work was funded by the Innovative Research Group Science Fund (no. Y020308133).

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Authors and Affiliations

  1. Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, People’s Republic of China

    Xuetuan Wei, Yuchun Xie, Liangrong Yang, Haojian Li, Lin Xu & Huizhou Liu

  2. National Key Laboratory of Biochemical Engineering, Chinese Academy of Sciences, Beijing, 100190, People’s Republic of China

    Xuetuan Wei, Yuchun Xie, Liangrong Yang, Haojian Li, Lin Xu & Huizhou Liu

  3. Graduate University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Xuetuan Wei & Lin Xu

  4. Technology Transfer Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Mingfang Luo

  5. No. 1 Bei Er Tiao, Zhong Guan Cun, Haidian District, Beijing, 100190, People’s Republic of China

    Mingfang Luo & Huizhou Liu

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  1. Xuetuan Wei
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  2. Mingfang Luo
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Correspondence to Mingfang Luo or Huizhou Liu.

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Wei, X., Luo, M., Xie, Y. et al. Strain Screening, Fermentation, Separation, and Encapsulation for Production of Nattokinase Functional Food. Appl Biochem Biotechnol 168, 1753–1764 (2012). https://doi.org/10.1007/s12010-012-9894-2

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  • DOI: https://doi.org/10.1007/s12010-012-9894-2

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