Biotechnology, Bioengineering and Applications of &lt;em&gt;Bacillus &lt;/em&gt;Nattokinase

Li, Yuan; Chen, Liangqi; Li, Jinyao

doi:10.20944/preprints202206.0402.v1

2022

DOI: 10.20944/preprints202206.0402.v1

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Biotechnology, Bioengineering and Applications of <em>Bacillus </em>Nattokinase

Yuan Li¹,

Liangqi Chen²,

Jinyao Li³

Abstract: Thrombosis has threatened human health in past decades. Bacillus nattokinase is a potential thrombolytic drug without side-effect and low cost and has been introduced into the consumer market as a functional food or dietary supplement. This review firstly summarizes the biodiversity sources and fermentation process of nattokinase, and systematically expounds the structure, cata-lytic mechanism and enzymatic properties of nattokinase; In view of the problems of low fermen-tation yield, insufficient activity and… Show more

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2023

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Cited by 4 publications

References 95 publications

(125 reference statements)

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Nattokinase: Insights into Biological Activity, Therapeutic Applications, and the Influence of Microbial Fermentation

Fang,

Yuan,

Wang

et al. 2023

Fermentation

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Nattokinase, a serine protease that originates from the traditional food natto, has garnered widespread attention due to its pharmacological functions and therapeutic potential. This review aims to delve into the major advancements of nattokinase across various domains, particularly its emerging roles in Alzheimer’s disease prevention and the treatment of retinal diseases, thereby seeking to usher in a newfound hope in the fields of neurology and ophthalmology. However, the production and preservation of nattokinase present a multitude of challenges, including issues of unstable yield and enzyme activity loss. To address these challenges, we explore potential solutions such as the heterologous expression of the nattokinase gene, the optimization of microbial fermentation strategies, and innovative purification methods. Furthermore, we focus on enhancing the stability and protection of nattokinase through encapsulation and immobilization techniques, thus ensuring its sustainability across a wide array of applications. This review provides readers with the opportunity to gain an in-depth understanding of the diverse prospects for nattokinase applications. Future research directions will encompass a deeper exploration of its biological mechanisms, the development of novel nattokinase derivatives, and the extension of its applications into a broader spectrum of disease treatments and health maintenance.

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Nattokinase: Insights into Biological Activity, Therapeutic Applications, and the Influence of Microbial Fermentation

Fang,

Yuan,

Wang

et al. 2023

Fermentation

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High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of Bacillus licheniformis

Li,

Yang,

Liu

et al. 2023

Fermentation

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During the production of nattokinase (NK) by Bacillus species, certain by-products such as poly-γ-glutamic acid (γ-PGA) are simultaneously synthesized. The impact of γ-PGA synthesis on NK production remains unclear. In this study, we knocked out the pgsC gene, a component of the γ-PGA synthetase cluster (pgsBCA), and constructed a novel recombinant strain, Bacillus licheniformis BL11. Next, we compared the fed-batch fermentation profiles of BL11 and its parental strain BL10, conducted transcriptional analysis, and measured intracellular ATP content. We also optimized glucose-feeding strategies under varying oxygen supply conditions. Our results indicated that the utilization rates of glucose and soybean meal were both improved in the pgsC-deficient strain BL11, and NK activity was enhanced. Furthermore, the transcriptional levels of genes involved in glycolysis and the TCA cycle were relatively upregulated in BL11. The maximal NK activity reached 2522.2 FU/mL at 54 h of fermentation using a constant glucose-feeding rate of 5.0 g/(L·h) under high oxygen supply conditions. The newly developed recombinant strain B. licheniformis BL11, along with the optimized feeding strategy, shows promise for large-scale NK production.

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Characterization of a Nattokinase from the Newly Isolated Bile Salt-Resistant Bacillus mojavensis LY-06

Chen

2022

Foods

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Nattokinase is a potential new thrombolytic drug because of its strong thrombolytic effect, high safety, and low cost. However, there is no research reporting on bile salt-tolerant nattokinase-producing probiotics. In this study, the bile salt-tolerant nattokinase-producing strain Bacillus mojavensis LY-06 was isolated from local Xinjiang douchi, and the fermentation yield of nattokinase of 1434.64 U/mL was obtained by both a single factor experiment and an orthogonal experiment. A gene responsible for fibrinolysis (aprY) was cloned from the genome of strain Bacillus mojavensis LY-06, and the soluble expression of this gene in Escherichia coli (rAprY, fused with His-tag at C-terminus) was achieved; molecular docking elucidates the cause of insoluble expression of rAprY. The optimal pH and temperature for the fibrinolysis activity of nattokinase AprY fermented by Bacillus mojavensis LY-06 were determined to be pH 6.0 and 50 °C, respectively. However, the optimal pH of rAprY expressed in Escherichia coli was 8, and its acid stability, thermal stability, and fibrinolytic activity were lower than those of AprY. Bioinformatics analysis found that the His-tag carried at the C-terminus of rAprY could affect its acidic stability by changing the isoelectric point and surface charge of the enzyme; in contrast to AprY, changes in the number of internal hydrogen bonds and the flexibility of the loop region in the structure of rAprY resulted in lower fibrinolytic activity and poorer thermal stability.

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Biotechnology, Bioengineering and Applications of <em>Bacillus </em>Nattokinase

Cited by 4 publications

References 95 publications

Nattokinase: Insights into Biological Activity, Therapeutic Applications, and the Influence of Microbial Fermentation

Nattokinase: Insights into Biological Activity, Therapeutic Applications, and the Influence of Microbial Fermentation

High Production of Nattokinase via Fed-Batch Fermentation of the γ-PGA-Deficient Strain of Bacillus licheniformis

Characterization of a Nattokinase from the Newly Isolated Bile Salt-Resistant Bacillus mojavensis LY-06

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