Strain Screening from Traditional Fermented Soybean Foods and Induction of Nattokinase Production in Bacillus subtilis MX-6

Man, Lili; Xiang, Dian-Jun; Zhang, Chunlan

doi:10.1007/s12602-017-9382-7

Cited by 26 publications

(19 citation statements)

References 25 publications

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“…Due to the high amino acid concentrations in TPW, the role of amino acid-induced enzyme production in TPW can be excluded. This standpoint is consistent with other researchers, such as Li-Li Man, who suggested that the peptide component in the soybean peptone was a critical factor in facilitating the synthesis of nattokinase rather than amino acids [17].…”

Section: Discussionsupporting

confidence: 92%

Tofu processing wastewater as a low-cost substrate for high activity nattokinase production using Bacillus subtilis

Zhan

Guo

et al. 2021

BMC Biotechnol

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Background Even though tofu is a traditional Chinese food loved by Asian people the wastewater generated during the production of tofu can pollute the environment, and the treatment of this generated wastewater can increase the operating cost of the plant. In this study, the production of nattokinase could be achieved by using the nitrogen source in tofu processing wastewater (TPW) instead of using the traditional nattokinase medium. This meets the need for the low-cost fermentation of nattokinase and at the same time addresses the environmental pollution concerns caused by the wastewater. Bacillus subtilis 13,932 is, a high yielding strain of nattokinase, which is stored in our laboratory. To increase the activity of nattokinase in the tofu process wastewater fermentation medium, the medium components and culture parameters were optimized. Nattokinase with high enzymatic activity was obtained in 7 L and 100 L bioreactors when TPW was used as the sole nitrogen source catalyzed by Bacillus subtilis. Such a result demonstrates that the production of nattokinase from TPW fermentation using B. subtilis can be implemented at an industrial level. Results The peptide component in TPW is a crucial factor in the production of nattokinase. Box–Behnken design (BBD) experiments were designed to optimize various critical components, i.e., Glucose, TPW, MgSO4·7H2O, CaCl2, in nattokinase fermentation media. A maximum nattokinase activity was recorded at 37 °C, pH 7.0, 70 mL liquid medium, and 200 rpm. The highest nattokinase activities obtained from 7 to 100 L bioreactors were 8628.35 ± 113.87 IU/mL and 10,661.97 ± 72.47 IU/mL, respectively. Conclusions By replacing the nitrogen source in the original medium with TPW, there was an increase in the enzyme activity by 19.25% after optimizing the medium and culture parameters. According to the scale-up experiment from conical flasks to 100 L bioreactors, there was an increase in the activity of nattokinase by 47.89%.

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Section: Discussionsupporting

confidence: 92%

Tofu processing wastewater as a low-cost substrate for high activity nattokinase production using Bacillus subtilis

Zhan

Guo

et al. 2021

BMC Biotechnol

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“…NK was extracted from traditional fermented food using B. subtilis natto (Sumi et al 1987) [17]. The effects of NK on thrombosis have been confirmed in previous studies.…”

Section: Discussionmentioning

confidence: 90%

Nattokinase Crude Extract Inhibits Hepatocellular Carcinoma Growth in Mice

Yan

Wang

Qian

et al. 2019

Journal of Microbiology and Biotechnology

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Nattokinase (NK, E.C. 3.4.21.62) is a serine protease produced by Bacillus subtilis natto that shows promise for the treatment of thrombotic disease. In this study, we assessed the effects of NK on the development of hepatocellular carcinoma (HCC), a principal malignancy of the liver that causes morbidity and mortality worldwide. Crude extracts of NK (NCE) were isolated from fermentation medium by centrifugation and separated into three fractions (<10 K, 100~30 K and >30K). Orthotopic HCC mouse models were established and NCE was administered by oral gavage. H&E staining was performed to examine the pathology of HCC livers. Immunohistochemistry and immunofluorescence were used to evaluate FOXM1, CD31, CD44 and vimentin expression in the liver. Compared to PBS groups, NCE increased the survival rates of HCC-bearing mice to 31% and decreased ascites. Low-intensity ultrasound imaging showed that the hypoechoic mass area was lower in NCE-treated mice and that tumor growth significantly decreased. IHC staining showed that the expression of FOXM1 was inhibited by NCE treatment. Immunofluorescence results revealed lower levels of CD31, CD44 and vimentin in the NCE groups. Taken together, these data demonstrate that NCE from Bacillus subtilis natto improves survival and inhibits tumor growth in HCC mice.

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“…A single dose of nattokinase administration showed the enhanced fibrinolysis and anticoagulation profiles (Kurosawa et al., 2015). Nattokinase is mainly produced by the Bacillus subtilin (natto) during the fermentation of soybeans or soybeans‐based ingredients to produce Natto‐containing foods (Cai et al., 2017; Man et al., 2019). Now, Nattokinase is typically used as a functional factor in health functional food to prevent cardiovascular diseases and thrombosis.…”

Section: Introductionmentioning

confidence: 99%

“…Natto-containing foods (Cai et al, 2017;Man et al, 2019). Now, Nattokinase is typically used as a functional factor in health functional food to prevent cardiovascular diseases and thrombosis.…”

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confidence: 99%

Microencapsulation of nattokinase from fermentation by spray drying: Optimization, comprehensive score, and stability

et al. 2021

Food Science & Nutrition

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Nattokinase from fermentation has recently gained more attention due to its beneficial effects on cardiovascular system. However, the instability of free nattokinase limits its application. The aim of the study was to develop a spray‐drying microencapsulation process to obtain the nattokinase powder with high activity, high quality, and strong storage stability. Hence, the microencapsulation process of nattokinase from fermentation by spray drying was optimized. Experiments of single‐factor and response surface methodology were used to assess the comprehensive scores and nattokinase activities. According to single‐factor and response surface methodology results, optimum parameters of microencapsulation process of the nattokinase power by spray drying were 30% of mass ratio of wall materials, 139°C of air inlet temperature, 8 L/h of feed rate, and 80°C of outlet temperature. The final optimized result encompassed a comprehensive score of 96, nattokinase activity of 1,340 IU/ml, and moisture content of 4.1 ± 0.1%. In addition, the microencapsulated nattokinase power showed strong storage stability in the conditions of different temperatures and pH. After 30 days of storage, the nattokinase powder was still white or light yellow, with a special smell, no peculiar smell and paste taste, and no impurity. These results build the basis of further industrialization of the nattokinase powder from fermentation broth by spray drying.

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Strain Screening from Traditional Fermented Soybean Foods and Induction of Nattokinase Production in Bacillus subtilis MX-6

Cited by 26 publications

References 25 publications

Tofu processing wastewater as a low-cost substrate for high activity nattokinase production using Bacillus subtilis

Tofu processing wastewater as a low-cost substrate for high activity nattokinase production using Bacillus subtilis

Nattokinase Crude Extract Inhibits Hepatocellular Carcinoma Growth in Mice

Microencapsulation of nattokinase from fermentation by spray drying: Optimization, comprehensive score, and stability

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