Enhanced production of nattokinase from UV mutated Bacillus sp.
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.
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.
Copyright (c) 2013 V. Mohanasrinivasan, C. Subathra Devi, Ritusree Biswas, Falguni Paul, Mohor Mitra, E. Selvarajan, V. Suganthi
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).