Enhanced production of nattokinase from UV mutated Bacillus sp.

Authors

  • V. Mohanasrinivasan Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • C. Subathra Devi Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • Ritusree Biswas Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • Falguni Paul Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • Mohor Mitra Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • E. Selvarajan Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014
  • V. Suganthi Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

DOI:

https://doi.org/10.3329/bjp.v8i2.13690

Keywords:

Modified Holmstorm, Nattokinase, Shrimp shell, Strain improvement

Abstract

In the recent years, nattokinase is one of the most-often employed among the several thrombolytic agents used clinically, particularly because of its lower cost comparing to other thrombolytic agents. In the present research work, Bacillus sp. was isolated from the heterogeneous microbial population present in the soil sample and screened for the production of nattokinase. The production of the enzyme was carried out using two different media (with and without shrimp shell substrate). Nattokinase activity (clot buster) was determined by using a modified Holmstorm method. The production strain SFN01 was improved by random mutagenesis (UV radiation) and the enzyme activity was checked with the enzyme produced by wild strain. The mutated strains had exhibited a higher clot lysis activity in which 1 unit of the enzyme completely lyses 1 mL of human blood when compared to the wild strain. Nattokinase produced by SFN showed a retention time of 10.6 min in RP-HPLC chromatogram.

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Author Biographies

V. Mohanasrinivasan, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

Assistant Professor Senior

C. Subathra Devi, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

Assistant Professor Senior

Ritusree Biswas, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

SBST

Falguni Paul, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

SBST

E. Selvarajan, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

Research Associate

V. Suganthi, Division of Environmental Biotechnology, School of Bio Sciences and Technology, VIT University, Vellore 632014

Research Associate

References

Besson F, Michel G. Mycosubtilins B and C: Minor antibiotics from mycosubtilin producer Bacillus subtilis. Int Microbiol. 1990; 62: 93-99.

Chang CT, Fan MH, Kuo FC, Sung HY. Potent fibrinolytic enzyme from a mutant of Bacillus subtilis IMR-NK1. J Agric Food Chem. 2000; 48: 3210-16.

Chung S, Kong H, Buyer JS, Lakshman DK, Lydon J, Kim SD, Roberts DP. Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soil borne pathogens of cucumber and pepper. Appl Microbiol Biotechnol. 2008; 80: 11523.

Dubey R, Kumar J, Agrawala D, Char T, Pusp P. Isolation, production, purification, assay and characterization of fibrinolytic enzymes (nattokinase, streptokinase and urokinase) from bacterial sources. Afr J Biotechnol. 2011; 10: 1408-20.

Fiddaman PJ, Rossall S. The production of antifungal volatiles by Bacillus subtilis. J Appl Bacteriol. 1993; 74: 119-26.

Fujita M, Nomura K, Hong K, Ito Y, Asada A, Nishimuro S. Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan. Biochem Biophys Res Commun. 1993; 197: 1340-47.

Fujita M, Hong K, Ito Y, Fuji R, Kariya K, Nishimuro S. Thrombolytic effect of nattokinaseon a chemically induced thrombosis model in rat. Biol Pharmaceut Bull. 1995b; 18: 1387-91.

Holmstrom B. Streptokinase assay on large agar diffusion plates. Acta Chem Scand. 1965; 19: 1549-54.

Kim W, Choi K, Kim Y, Park H, Choi J, Lee Y, Oh H, Kwon I, Lee S. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkook-Jang. Appl Environ Microbiol. 1996; 62: 2482-88.

Klich MA, Lax AR, Bland JM. Inhibition of some mycotoxigenic fungi by iturin A, a peptidolipid produced by Bacillus subtilis. Mycopathologia 1991; 116: 77-80.

Peng Y, Huang Q, Zhang RH, Zhang YZ. Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food. Comp Biochem Physiol. Part B: Biochem Molecular Biol. 2003; 134: 45-52.

Reddy KS. India wakes up to threat of cardiovascular disease. J Am Coll Cardiol. 2007; 50: 1370-72.

Sumi H, Hamada H, Nakanishi K, Hiratani H. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematologica. 1990; 84: 139-43.

Sumi H, Hamada H, Tsushima H, Mihara H, Muriki H. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese natto: A typical and popular soybean food in the Japanese diet. Experientia. 1987; 43: 1110-11.

Thaer TAA, Adinarayana K, Poluri E. Isolation and mutagenesis of Streptokinase producing bacteria. Am J Immunol. 2005; 4: 125-29.

Urano T, Ihara H, Umemura K, Suzuki Y, Oike M, Akita S, Tsukamoto Y, Suzuki I, Takada A. The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis cleaves and inactivates plasminogen activator inhibitor type 1. J Biol Chem. 2001; 276: 24690-96.

Wang C, Du M, Zheng D, Kong F, Zu G, Feng Y. Purification and characterization of nattokinase from Bacillus subtilis Natto B-12. J Agric Food Chem. 2009; 57: 9722-29.

Published

2013-03-07

How to Cite

Mohanasrinivasan, V., C. Subathra Devi, R. Biswas, F. Paul, M. Mitra, E. Selvarajan, and V. Suganthi. “Enhanced Production of Nattokinase from UV Mutated Bacillus Sp”. Bangladesh Journal of Pharmacology, vol. 8, no. 2, Mar. 2013, pp. 110-5, doi:10.3329/bjp.v8i2.13690.

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Research Articles