Advances in Nattokinase Research: Therapeutic Potential and Applications | Wuhan University Journal of Natural Sciences

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Table 1

Overview of enzyme activity differences between different fermentation methods

Fermentation category	Optimization method	Fermentation strain	Specific conditions	NK enzyme activity	Reference
Solid state fermentation	Univariate and response surface tests	Bacillus natto JNFE 0127	Inoculum 6%, fermentation at 30 °C for 24 h, with KCl 0.1%, sucrose 2%, tryptone 1.5%	9 924.81 IU/g	[62]
	Univariate and orthogonal tests	Bacillus subtilis BS21076	Soybeans baked and fried at 90 °C for 5 min as materials, added 3% konjac powder, inoculum 8%, fermentation at 37 °C for 24 h	(3 582.48±83.13) IU/g	[63]
	Univariate and response surface tests	Bacillus natto	Half of the crushed soybeans used as materials, inoculum 11%, fermentation at initial pH for 45 h	1 408 IU/g	[64]
	Univariate and response surface tests	Bacillus subtilis SWS01	Initial moisture content 55%, 90 g/250 mL dry soybean meal, inoculum 6%, initial pH, fermentation at 36 °C for 25 h	5 421 IU/g	[65]
Two-species solid state fermentation	Single factor test	Bacillus natto F-2-4 Bacillus natto S-15	Inoculation volume ratio of two strains 1:1, inoculum 4%	6 287.16 IU/g	[66]
Liquid fermentation	Univariate and orthogonal tests	Bacillus natto NK4	Inoculum 2%, initial medium pH 6.0, fermentation at 34 °C for 72 h	848.28 IU/mL	[67]
	Univariate and orthogonal tests	Bacillus natto WTC016	Fermentation at 30 °C, medium pH 7.0, inoculum 2%, sample volume 60 mL, fermentation time 26 h, loading amount 24%	(3 284±58) IU/mL	[68]
	Univariate and response surface tests	Bacillus subtilis X3	Fermentation at 34 °C, medium pH 6.5, inoculum 2%, loading 20%, fermentation time 30 h	393.095 IU/mL	[69]
Liquid fermentation	Univariate and response surface tests	Bacillus natto	Fermentation at 34 °C, medium pH 7.5, inoculum 3%, fermentation time 95 h, bacterial age 17.5 h	5 087 IU/mL	[70]
	Single factor test	Recombinant Bacillus subtilis strain PSP2	Fermentation medium pH 7.0, containing 1.0% trypsin, 1.0% oyster protein hydrolysate, 2.0% maltose, and 0.5% sodium chloride, fermentation time 72 h	(390.23±10.24) IU/mL	[71]
	Univariate and response surface tests	Recombinant Bacillus subtilis strain 13932	Fermentation medium: glycerol 23 g/L, soybean residue 96 g/L, MgSO₄·7H₂O 0.8 g/L, CaCl₂ 0.7 g/L, initial pH 7.5, fermentation at 37 °C, inoculum 7%	(10 576.28 ± 91.78) IU/mL	[72]
	Univariate and orthogonal tests	Bacillus natto ACCC10614	Fermentation medium: 2% soybean meal, pH 7.0, inoculum 1%, fermentation time 48 h	4 429.6 IU/mL	[73]
Mixed liquid fermentation of three strains	Univariate and orthogonal tests	Mutagenic Bacillus amyloliquefaciens CBL-2-7-10-6, Mutagenesis of Bacillus subtilis DNS-17-3-15-7 and L-5-8-15	Fermentation medium: 4% glucose, 4% soy peptone, 0.04% MgSO₄, 0.06% CaCl₂, 0.2% K₂HPO₄, 0.1% KH₂PO₄, pH 7.0. The inoculations of CBL-2-7-10-6, DNS-17-3-15-7 and L-5-8-15 were 670, 670 and 1 000 μL/100 mL, fermentation temperature 31 ℃, fermentation time 50 h	4 952.33 IU/mL	[74]

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