Study on the mechanism of production of γ-PGA and nattokinase in Bacillus subtilis natto based on RNA-seq analysis

Li, Min; Zhang, Zilong; Li, Shenwei; Tian, Zhen; Ma, Xinbin

doi:10.1186/s12934-021-01570-x

Cited by 30 publications

(22 citation statements)

References 55 publications

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“… B. subtilis PGA production was measured using a cetryltrimethylammonium bromide (CTAB) assay as previously described ( 38 , 39 ). Cells were grown in LB broth containing various concentrations of ZnSO 4 or CuSO 4 in LB media overnight at 37°C for 16–18 h. Cells were then pelleted from 15 mL of culture by centrifugation at 10000 × g for 30 min at 4°C and the supernatant was retained.…”

Section: Methodsmentioning

confidence: 99%

Poly-Gamma-Glutamic Acid Secretion Protects Bacillus subtilis from Zinc and Copper Intoxication

et al. 2022

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Zinc and copper are potent antimicrobial compounds. As such, bacteria have evolved a diverse range of tools to prevent metal intoxication. Here, we show that the Gram-positive model organism, Bacillus subtilis , produces poly-gamma-glutamic acid (γ-PGA) as a protective mechanism against zinc and copper intoxication and that zinc and copper dependent γ-PGA production occurs by a yet undefined mechanism independent of known γ-PGA regulation pathways.

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Section: Methodsmentioning

confidence: 99%

Poly-Gamma-Glutamic Acid Secretion Protects Bacillus subtilis from Zinc and Copper Intoxication

et al. 2022

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“…The isolated supernatants were dotted into the middle of a 0.5% agarose plate containing thrombin and fibrinogen solution at pH 7.4. The diameters (mm) of the clear zones indicated the thrombolytic activity [15]. The positive-control activity was measured with plasmin.…”

Section: Methodsmentioning

confidence: 99%

“…Chungkookjang is comparable to Japanese natto, soybeans fermented with B. subtilis natto for approximately two days. During the fermentation of soybeans, B. subtilis natto produces nattokinase and γ-PGA, which help prevent or treat cardiovascular diseases by improving hypertension and dyslipidemia and reducing blood clotting [15]. B. subtilis, B. licheniformis, and B. amyloliquefaciens also have fibrinolytic activities and produce γ-PGA, but different strains have different efficiencies [7,8].…”

Section: Introductionmentioning

confidence: 99%

Alleviation of Neuronal Cell Death and Memory Deficit with Chungkookjang Made with Bacillus amyloliquefaciens and Bacillus subtilis Potentially through Promoting Gut–Brain Axis in Artery-Occluded Gerbils

Zhang

Ryu

et al. 2021

Foods

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Short-term fermented soybeans (chungkookjang) with specific Bacillus (B.) spp. have anti-obesity, antidiabetic, and anti-stroke functions. We examined the hypothesis that the long-term consumption of B. amyloliquefaciens SCGB 1 fermented (CKJ1) and B. subtilis SCDB 291 (CKJ291) chungkookjang can alleviate clinical symptoms and hyperglycemia after ischemic stroke by promoting the gut microbiota–brain axis. We examined this hypothesis in Mongolian male gerbils with stroke symptoms induced by carotid artery occlusion. The artery-occluded gerbils were divided into five groups: no supplementation (Control, Normal-control), 4% cooked soybeans (CSB), CKJ1, or CKJ291 in a high-fat diet for 3 weeks. The carotid arteries of gerbils in the Control, CSB, CKJ1, and CKJ291 groups were occluded for 8 min and they then continued on their assigned diets for an additional 3 weeks. Normal-control gerbils had no artery occlusion. The diets in all groups contained an identical macronutrient composition using starch, casein, soybean oil, and dietary fiber. The CSB, CKJ1, and CKJ291 groups exhibited less neuronal cell death than the Control group, while the CKJ1 group produced the most significant reduction among all groups, as much as 85% of the Normal-control group. CKJ1 and CKJ291 increased the blood flow and removal of blood clots, as determined by Doppler, more than the Control. They also showed more improvement in neurological disorders from ischemic stroke. Their improvement showed a similar tendency as neuronal cell death. CKJ1 treatment improved memory impairment, measured with Y maze and passive avoidance tests, similar to the Normal-control. The gerbils in the Control group had post-stroke hyperglycemia due to decreased insulin sensitivity and β-cell function and mass; the CKJ291, CSB, and CKJ1 treatments protected against glucose disturbance after artery occlusion and were similar to the Normal-control. CKJ1 and CKJ291 also reduced serum tumor necrosis factor-α concentrations and hippocampal interleukin-1β expression levels, compared to the Control. CKJ1 and CKJ291 increased the contents of Lactobacillus, Bacillus, and Akkermansia in the cecum feces, similar to the Normal-control. Picrust2 analysis showed that CKJ1 and CKJ291 increased the propionate and butyrate metabolism and the starch and glucose metabolism but reduced the lipopolysaccharide biosynthesis and fatty acid metabolism compared to the Control. In conclusion, daily CKJ1 and CKJ291 intake prevented neuronal cell death and memory dysfunction from the artery occlusion by increasing blood flow and β-cell survival and reducing post-stroke-hyperglycemia through modulating the gut microbiome composition and metabolites to influence the host metabolism, especially inflammation and insulin resistance, protecting against neuronal cell death and brain dysfunction. CKJ1 had better effects than CKJ291.

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“…However, the effect of this by-product on the fermentation of nattokinase is still controversial. Li et al found that the activity of nattokinase was the highest when the production of γ-PGA reached the maximum value; thus Li et al speculated that the γ-PGA produced by Bacillus subtilis during the fermentation process could stimulate the biosynthesis of nattokinase [ 45 ]. Contrarily, Wei et al believed that γ-PGA mixed in nattokinase was an immunogenic substance that would cause allergic reactions in humans that consume it.…”

Section: Study On Fermentation Process Of Nattokinasementioning

confidence: 99%

Biotechnology, Bioengineering and Applications of Bacillus Nattokinase

Chen

2022

Biomolecules

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Thrombosis has threatened human health in past decades. Bacillus nattokinase is a potential low-cost thrombolytic drug without side-effects and has been introduced into the consumer market as a functional food or dietary supplement. This review firstly summarizes the biodiversity of sources and the fermentation process of nattokinase, and systematically elucidates the structure, catalytic mechanism and enzymatic properties of nattokinase. In view of the problems of low fermentation yield, insufficient activity and stability of nattokinase, this review discusses the heterologous expression of nattokinase in different microbial hosts and summarizes the protein and genetic engineering progress of nattokinase-producing strains. Finally, this review summarizes the clinical applications of nattokinase.

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Study on the mechanism of production of γ-PGA and nattokinase in Bacillus subtilis natto based on RNA-seq analysis

Cited by 30 publications

References 55 publications

Poly-Gamma-Glutamic Acid Secretion Protects Bacillus subtilis from Zinc and Copper Intoxication

Poly-Gamma-Glutamic Acid Secretion Protects Bacillus subtilis from Zinc and Copper Intoxication

Alleviation of Neuronal Cell Death and Memory Deficit with Chungkookjang Made with Bacillus amyloliquefaciens and Bacillus subtilis Potentially through Promoting Gut–Brain Axis in Artery-Occluded Gerbils

Biotechnology, Bioengineering and Applications of Bacillus Nattokinase

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