Construction of a 3D model of nattokinase, a novel fibrinolytic enzyme from Bacillus natto

Zheng, Zhaohui; Zuo, Zhenyu; Liu, Zhigang; Tsai, Keng‐Chang; Liu, Aifu; Zou, Guolin

doi:10.1016/j.jmgm.2004.10.002

Cited by 51 publications

(38 citation statements)

References 28 publications

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“…The two a-helices formed by non-conserved segments are not critical. The different conformation and the slight decrease in the catalytic efficiency reveal that substitution of Ile 11 and Gly 34 alters the secondary structure but does not significantly affect the catalytic triad. Therefore, both amino acids are believed to participate in hydrogen bond formation with residues around the catalytic triad except for His 32 , Asp 64 and Ser 221 .…”

Section: Conserved Regions Of Propeptide Fold Into Three B-strands Thmentioning

confidence: 95%

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Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase

et al. 2010

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a b s t r a c tHere, we show that during in vivo folding of the precursor, the propeptide of subtilisin nattokinase functions as an intramolecular chaperone (IMC) that organises the in vivo folding of the subtilisin domain. Two residues belonging to b-strands formed by conserved regions of the IMC are crucial for the folding of the subtilisin domain through direct interactions. An identical protease can fold into different conformations in vivo due to the action of a mutated IMC, resulting in different kinetic parameters. Some interfacial changes involving conserved regions, even those induced by the subtilisin domain, blocked subtilisin folding and altered its conformation. Insight into the interaction between the subtilisin and IMC domains is provided by a three-dimensional structural model.

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Section: Conserved Regions Of Propeptide Fold Into Three B-strands Thmentioning

confidence: 95%

“…We have previously used site-directed mutagenesis and molecular dynamic simulations to probe the importance of hydrogen bonds in the active site of NK, and we constructed a three-dimensional model of NK [11,12]. In addition, our previous study demonstrated that active NK was not detected without the assistance of the propeptide [13].…”

Section: Introductionmentioning

confidence: 99%

Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase

et al. 2010

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“…Over the last 10 years, several effective thrombolytic agents have been identified and characterized from microorganisms [4][5][6], earthworms [7], snake venoms [8,9], centipede venoms [10], insects [11], and leeches [12]. The agents are of interest as useful tools for understanding fibrinolytic mechanism and as potential therapeutic drugs.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of a fibrinolytic enzyme of Bacillus subtilis DC33, isolated from Chinese traditional Douchi

Wang

Лі

et al. 2006

J IND MICROBIOL BIOTECHNOL

148

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Bacillus subtilis DC33 producing a novel fibrinolytic enzyme was isolated from Ba-bao Douchi, a traditional soybean-fermented food in China. The strong fibrin-specific enzyme subtilisin FS33 was purified to electrophoretic homogeneity using the combination of various chromatographic steps. The optimum temperature, pH value, and pI of subtilisin FS33 were 55°C, 8.0, and 8.7, respectively. The molecular weight was 30 kDa measured by SDS-PAGE under both reducing and non-reducing conditions. The enzyme showed a level of fibrinolytic activity that was about six times higher than that of subtilisin Carlsberg. The first 15 amino acid residues of N-terminal sequence of the enzyme were A-Q-S-V-P-Y-G-I-P-Q-I-K-A-P-A, which are different from that of other known fibrinolytic enzymes. The amidolytic activities of subtilisin FS33 were inhibited completely by 5 mM phenylmethanesulfonyl fluoride (PMSF) and 1 mM soybean trypsin inhibitor (SBTI), but 1,4-dithiothreitol (DTT), b-mercaptoethanol, and p-hydroxymercuribenzoate (PHMB) did not affect the enzyme activity; serine and tryptophan are thus essential in the active site of the enzyme. The highest affinity of subtilisin FS33 was towards N-Succ-Ala-Ala-Pro-PhepNA. Therefore, the enzyme was considered to be a subtilisin-like serine protease. The fibrinolytic enzyme had a high degrading activity for the Bb-chains and Aa-chain of fibrin(ogen), and also acted on thrombotic and fibrinolytic factors of blood, such as plasminogen, urokinase, thrombin, and kallikrein. So subtilisin FS33 was able to degrade fibrin clots in two ways, i.e., (a) by forming active plasmin from plasminogen and (b) by direct fibrinolysis.

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“…As compared to the clinical thrombolytic drugs (urokinase and streptokinase), nattokinase (NK) with many advantages such as confirmed efficacy, prolonged effects, cost effectiveness and preventative use is a potent thrombolytic enzyme in vitro and in vivo (Zheng et al 2005). For example, it has four times fibrinolytic activity over plasmin (Sumi et al 1992), and the activity can be retained in the blood for more than 3 h (Fujita et al 1995).…”

Section: Introductionmentioning

confidence: 99%

Co-Production of Nattokinase and Poly (γ-Glutamic Acid) Under Solid-State Fermentation Using Soybean and Rice Husk

Nie

Zhu

Liu

et al. 2015

Braz. arch. biol. technol.

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Construction of a 3D model of nattokinase, a novel fibrinolytic enzyme from Bacillus natto

Cited by 51 publications

References 28 publications

Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase

Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase

Purification and characterization of a fibrinolytic enzyme of Bacillus subtilis DC33, isolated from Chinese traditional Douchi

Co-Production of Nattokinase and Poly (γ-Glutamic Acid) Under Solid-State Fermentation Using Soybean and Rice Husk

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