Strain Screening, Fermentation, Separation, and Encapsulation for Production of Nattokinase Functional Food

This study presents a novel and integrated preparation technology for nattokinase functional food, including strain screening, fermentation, separation, and encapsulation. To rapidly screen a nattokinase-productive strain, PCR-based screening method was combined with fibrinolytic activity-based meth...

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 2012-12, Vol.168 (7), p.1753-1764
Main Authors: Wei, Xuetuan, Luo, Mingfang, Xie, Yuchun, Yang, Liangrong, Li, Haojian, Xu, Lin, Liu, Huizhou
Format: Article
Language:English
Subjects:
Acrylates - chemistry
Amino Acid Sequence
Bacillus subtilis
Bacillus subtilis - isolation & purification
Bacillus subtilis - metabolism
Base Sequence
Biochemistry
Biological and medical sciences
Biotechnology
Capsules
Chemical Precipitation
Chemistry
Chemistry and Materials Science
Cicer - enzymology
Cicer - microbiology
Cicer arietinum
Enzyme Stability
Ethanol - chemistry
Ethanol - isolation & purification
Ethanol - metabolism
Fermentation
Fibrinolysis
Functional foods & nutraceuticals
Fundamental and applied biological sciences. Psychology
Gelatin - chemistry
Glycine max - enzymology
Glycine max - microbiology
Hydrogen-Ion Concentration
Kinases
Methods. Procedures. Technologies
Microbial engineering. Fermentation and microbial culture technology
Molecular Sequence Data
Polymerase Chain Reaction
Polymethacrylic Acids - chemistry
Subtilisins - biosynthesis
Subtilisins - chemistry
Subtilisins - genetics
Subtilisins - metabolism
Time Factors
Water - chemistry
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Summary:This study presents a novel and integrated preparation technology for nattokinase functional food, including strain screening, fermentation, separation, and encapsulation. To rapidly screen a nattokinase-productive strain, PCR-based screening method was combined with fibrinolytic activity-based method, and a high productive strain, Bacillus subtilis LSSE-22, was isolated from Chinese soybean paste. Reduction of poly-γ-glutamic acid (γ-PGA) concentration may contribute to separation of nattokinase and reduction of late-onset anaphylaxis risk. Chickpeas were confirmed as the favorable substrate for enhancement of nattokinase production and reduction of γ-PGA yield. Using cracked chickpeas, the nattokinase activity reached 356.25 ± 17.18 FU/g (dry weight), which is much higher than previous reports. To further reduce γ-PGA concentration, ethanol fractional extraction and precipitation were applied for separation of nattokinase. By extraction with 50 % and precipitation with 75 % ethanol solution, 4,000.58 ± 192.98 FU/g of nattokinase powders were obtained, and the activity recovery reached 89 ± 1 %, while γ-PGA recovery was reduced to 21 ± 2 %. To improve the nattokinase stability at acidic pH condition, the nattokinase powders were encapsulated, and then coated with methacrylic acid–ethyl acrylate copolymer. After encapsulation, the nattokinase was protected from being denatured under various acid conditions, and pH-responsible controlled release at simulated intestinal fluid was realized.
ISSN:0273-2289
1559-0291
DOI:10.1007/s12010-012-9894-2