Screening for Optimal Parameters of Nattokinase Synthesis by Bacillus subtilis natto in Solid-State Fermentation

Cuong Viet Bui, Minh Nguyet Thi Nguyen, Minh Hanh Thi Truong, Xuan Dong Bui

Abstract


Nattokinase, which is an extracellular enzyme synthesized by Bacillus subtilis natto, is a medication for cardiovascular disease treatment. In this study, soybean seed was used as a substrate for culturing Bacillus subtilis natto in solid-state fermentation to produce nattokinase. The optimal culture parameters for solid-state fermentation to synthesize nattokinase by Bacillus subtilis natto were 2:50 (v:w) of the ratio of Bacillus subtilis natto pre-culture to the substrate, 42 h of fermentation time, and 3 cm of the thickness of the substrate. At this optimal culture parameter of solid-state fermentation by using Bacillus subtilis natto, the enzymatic activity of crude nattokinase was 7.13 ± 0.2, 16.24 ± 0.33, and 16.55 ± 0.06 (specific activity/mL), respectively. Furthermore, we aim to apply this study with large scale production initially. Thus, the ratio of reusing Bacillus subtilis natto in fermentation product to substrate for a new process of solid-state fermentation and circulation time of reusing Bacillus subtilis natto in new solid fermentation process were screened. The maximal enzymatic activity of crude nattokinase of 14.10 ± 0.18 (specific activity/mL) was found at 3:100 (w:w) of reusing Bacillus subtilis natto in fermentation product to substrate for a new process of solid-state fermentation. The suggestion for the circulation time of reusing Bacillus subtilis natto for new solid fermentation process was 2nd. The results of this study had provided the necessary information for further research on nattokinase.

Keywords


nattokinase; Bacillus subtilis natto; optimal culture parameters; reusing Bacillus subtilis natto; circulation time; solid-state fermentation.

Full Text:

PDF

References


Jeong, S.-J., et al., Characterization of AprE176, a fibrinolytic enzyme from Bacillus subtilis HK176, J. Microbiol. Biotechnol, 2015, 25(1), pp. 89-97.

Afifah, D.N., M. Sulchan, and D. Syah, Purification and characterization of a fibrinolytic enzyme from Bacillus pumilus 2. g isolated from Gembus, an Indonesian fermented food, Preventive nutrition and food science, 2014, 19(3), pp. 213.

WHO., Global status report on noncommunicable diseases 2010, Geneva, Switzerland, World Health Organization, 2011.

Mathers CD, L.D., Projections of global mortality and burden of disease from 2002 to 2030, PLoS Med, 3: e442., 2006.

Voet D, V.J., Biochemistry, 2nd ed, New York, USA: John Wiley and Sons, 1990.

Killer, M., et al., Current endovascular treatment of acute stroke and future aspects, Drug discovery today, 2010, 15(15), pp. 640-647.

Wang, C., et al., Purification and characterization of nattokinase from Bacillus subtilis Natto B-12, Journal of agricultural and food chemistry, 2009, 57(20), pp. 9722-9729.

Jo, H.-D., et al., Cloning and overexpression of aprE3-17 encoding the major fibrinolytic protease of Bacillus licheniformis CH 3-17, Biotechnology and Bioprocess Engineering: BBE, 2011, 16(2), pp. 352.

Peng, Y., et al., Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food, Comparative biochemistry and physiology part b: biochemistry and molecular biology, 2003, 134(1), pp. 45-52.

Sumi, H., et al., Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinases, Acta haematologica, 1990, 84(3), pp. 139-143.

Sizer, I.W., Medical Applications of Microbial Enzymes, Advances in Applied Microbiology, 1972, 15, pp. 1-11.

Underkofler, L., R. Barton, and S. Rennert, Production of microbial enzymes and their applications, Applied microbiology, 1958, 6(3), pp. 212.

Chand, S. and P. Mishra, Research and application of microbial enzymes, India’s contribution, in Biotechnology in India II, 2003, pp. 95-124.

Pandey, A., et al., Solid-state fermentation for the production of industrial enzymes, Current science, 1999, pp. 149-162.

Stanbury, P.F., A. Whitaker, and S.J. Hall, Principles of fermentation technology, Elsevier, 2013.

Subramaniyam, R. and R. Vimala, Solid-state and submerged fermentation for the production of bioactive substances: a comparative study, Int J Sci Nat, 2012, 3, pp. 480-486.

El-Shemy, H., Soybean and nutrition, Tech. Publish, Rijeka, Croatia, 2011.

A.O.A.C, Official Methods of Analysis of AOAC International, ed. 17th, Maryland: AOAC International, 2000.

Inc, A.E., Protease (Aspergillus llkaline proteinase, EC 3.4.21.63), Amano, pp. 77-78.

Montgomery, D.C. and G.C. Runger, Applied statistics and probability for engineers, John Wiley & Sons, 2010.

Saxena, R. and R. Singh, Amylase production by solid-state fermentation of agro-industrial wastes using Bacillus sp, Brazilian Journal of Microbiology, 2011, 42(4), pp. 1334-1342.

Kapoor, R. and B.P. Panda, Production of nattokinase from Bacillus sp. by solid-state fermentation, International Journal of Medical Sciences and Biotechnology, 2013, I, pp. 61-65.

Kellner, R., F. Lottspeich, and H.E. Meyer, Microcharacterization of proteins. 2008: John Wiley & Sons.

Liu, C., Y. Zhao, and W. Cao, Optimization of fermentation process on a fibrinolytic enzyme from Bacillus subtilis SBS, Biotechnology, 2003, 14(6), pp. 52-54.




DOI: http://dx.doi.org/10.18517/ijaseit.10.3.5492

Refbacks

  • There are currently no refbacks.


Published by INSIGHT - Indonesian Society for Knowledge and Human Development