Login Register Cart 0
Generic placeholder image

Cardiovascular & Hematological Agents in Medicinal Chemistry

Editor-in-Chief

ISSN (Print): 1871-5257
ISSN (Online): 1875-6182

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Production of Nattokinase from Bacillus amyloliquefaciens MRS18: A Bacterial Strain Isolated from Fermented Beans

Author(s): Simran B. Coutinho, Rutuja R. Shirodkar, Anju Kanjirakkandy, Vijayaganapathi Arulmozhi, Mohanasrinivasan Vaithilingam* and Subathra D. Chandrasekaran

Volume 22, Issue 2, 2024

Published on: 01 January, 2024

Page: [223 - 229] Pages: 7

DOI: 10.2174/0118715257275639231219054353

Price: $65

Abstract

Background: Nattokinase (NK) is a naturally occurring fibrinolytic protease enzyme obtained from the traditional Japanese food called Natto and has several uses in the pharmaceutical and medical industries. Nowadays, the most often used thrombolytic agent in the clinical field is NK, in part because it is less expensive than other thrombolytic medicines.

Objectives: The objective of this study is to investigate the screening, isolation and characterization of the NK enzyme-producing Bacillus strain from fermented Soya beans.

Methods: The sample of fermented soya beans were tested for the presence of fibrinolytic protease- producing bacteria, followed by the screening, extraction, characterization and clot lysis assays.

Results: A total of three isolates were screened for caseinolytic activities by casein hydrolysis assay. Out of those isolates, MRS18 was found to be potent in producing the enzyme proteinase. To determine the taxonomy and phylogeny of these isolates, biochemical and molecular characterization has been carried out. Bacillus amyloliquefaciens MRS18 has been found with the highest caseinolytic activity. The clot lysing ability of the potent strain Bacillus amyloliquefaciens was found to be 61.7% after 120 min, and on further purification, by ammonium sulphate precipitation method, the lysis percentage was found to be 656% after 120 min.

Conclusion: From the results of the present study, we concluded that Bacillus amyloliquefaciens isolated from the fermented soya beans produced an NK enzyme that exhibits immense potential to lyse blood clots.

« Previous Next »
Graphical Abstract

[1]
Mendis, S. Global atlas on cardiovascular disease prevention and control; World Health Organization, 2011.
[2]
Seattle, W. GBD compare data visualization. 2017. Available from: https://www.healthdata.org/data-tools-practices/interactive-visuals/gbd-compare
[3]
Jamali, N. Nattokinase: Structure, applications and sources. Biocatal. Agric. Biotechnol., 2022, 47, 102564.
[4]
Kundu, J.; Kundu, S. Cardiovascular disease (CVD) and its associated risk factors among older adults in India: Evidence from LASI Wave 1. Clin. Epidemiol. Glob. Health, 2022, 13, 100937.
[http://dx.doi.org/10.1016/j.cegh.2021.100937]
[5]
Rengaswamy, D.; Abdul Rahim, P. Fibrinolytic enzyme-an overview. Curr. Pharm. Biotechnol., 2022, 23(11), 1336-1345.
[http://dx.doi.org/10.2174/1389201023666220104143113] [PMID: 34983344]
[6]
Selvarajan, E.; Bhatnagar, N. Nattokinase: An updated critical review on challenges and perspectives. Cardiovasc. Hematol. Agents Med. Chem., 2017.
[7]
Kotb, E. The biotechnological potential of fibrinolytic enzymes in the dissolution of endogenous blood thrombi. Biotechnol. Prog., 2014, 30(3), 656-672.
[http://dx.doi.org/10.1002/btpr.1918] [PMID: 24799449]
[8]
Jeong, Y.K.; Park, J.U.; Baek, H.; Park, S.H.; Kong, I.S.; Kim, D.W.; Joo, W.H. Purification and biochemical characterization of a fibrinolytic enzyme from Bacillus subtilis BK-17. World J. Microbiol. Biotechnol., 2001, 17(1), 89-92.
[http://dx.doi.org/10.1023/A:1016685411809]
[9]
Weng, Y.; Yao, J.; Sparks, S.; Wang, K. Nattokinase: An oral antithrombotic agent for the prevention of cardiovascular disease. Int. J. Mol. Sci., 2017, 18(3), 523.
[http://dx.doi.org/10.3390/ijms18030523] [PMID: 28264497]
[10]
Collen, D.; Lijnen, H. Basic and clinical aspects of fibrinolysis and thrombolysis. Blood, 1991, 178(12), 3114-3124.
[11]
Kumar, S.S.; Sabu, A. Fibrinolytic enzymes for thrombolytic therapy. Adv. Exp. Med. Biol., 2019, 1148, 345-381.
[http://dx.doi.org/10.1007/978-981-13-7709-9_15]
[12]
Kotb, E. Microbial fibrinolytic enzyme production and applications.Microbial Functional Foods and Nutraceuticals; John Wiley & Sons Ltd., 2017, pp. 175-213.
[13]
Merlyn Keziah, S.; Subathra Devi, C. Molecular identification of autochthonous bacterial fibrinolytic protein producers from fermentative food preparations. Indian J. Biotechnol., 2021, 20, 154-162.
[14]
Dubey, R. Isolation, production, purification, assay and characterization of fibrinolytic enzymes (Nattokinase, Streptokinase and Urokinase) from bacterial sources. Afr. J. Biotechnol., 2011, 10(8), 1408-1420.
[15]
Brown, J.H. Bergey’s manual of determinative bacteriology; American Public Health Association, 1939.
[16]
Adhoni, S.A.; Shivasharana, C.; Kaliwal, B. Identification and characterisation of chlorella vulgaris for biodiesel production. Int. J. Scient. Res. Eng. Stud, 2016, 3(1)
[17]
Holmström, B.; Seppälä, P.; Vilhunen, R.; Enari, T-M. Streptokinase assay on large agar diffusion plates. Acta Chem. Scand., 1965, 19(7), 1549-1554.
[http://dx.doi.org/10.3891/acta.chem.scand.19-1549] [PMID: 4955306]
[18]
Jo, H.D.; Lee, H.A.; Jeong, S.J.; Kim, J.H. Purification and characterization of a major fibrinolytic enzyme from Bacillus amyloliquefaciens MJ5-41 isolated from Meju. J. Microbiol. Biotechnol., 2011, 21(11), 1166-1173.
[http://dx.doi.org/10.4014/jmb.1106.06008] [PMID: 22127128]
[19]
Nwagu, T.N.; Ugwuodo, C.J.; Onwosi, C.O.; Inyima, O.; Uchendu, O.C.; Akpuru, C. Evaluation of the probiotic attributes of Bacillus strains isolated from traditional fermented African locust bean seeds (Parkia biglobosa), “daddawa. Ann. Microbiol., 2020, 70(1), 20.
[http://dx.doi.org/10.1186/s13213-020-01564-x]
[20]
Ouoba, L.I.I.; Thorsen, L.; Varnam, A.H. Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments. Int. J. Food Microbiol., 2008, 124(3), 224-230.
[http://dx.doi.org/10.1016/j.ijfoodmicro.2008.03.026] [PMID: 18474404]
[21]
Odunfa, S.A.; Adeyele, S. Microbiological changes during the traditional production of ogi-baba, a West African fermented sorghum gruel. J. Cereal Sci., 1985, 3(2), 173-180.
[http://dx.doi.org/10.1016/S0733-5210(85)80027-8]
[22]
Srinivasan, V.M. Screening for staphylokinase producing Staphylococcus spp. from bovine milk sample. Int. J. Pharm. Pharm. Sci., 2013, 5(2), 601.
[23]
Devi, C.S.; Mohanasrinivasan, V.; Sharma, P.; Das, D.; Vaishnavi, B.; Naine, S.J. Production, purification and stability studies on nattokinase: A therapeutic protein extracted from mutant Pseudomonas aeruginosa CMSS isolated from bovine milk. Int. J. Pept. Res. Ther., 2016, 22(2), 263-269.
[http://dx.doi.org/10.1007/s10989-015-9505-5]
[24]
Sumi, H.; Hamada, H.; Nakanishi, K.; Hiratani, H. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase. Acta Haematol., 1990, 84(3), 139-143.
[http://dx.doi.org/10.1159/000205051] [PMID: 2123064]
[25]
Hsia, C.H.; Shen, M.C.; Lin, J.S.; Wen, Y.K.; Hwang, K.L.; Cham, T.M.; Yang, N.C. Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects. Nutr. Res., 2009, 29(3), 190-196.
[http://dx.doi.org/10.1016/j.nutres.2009.01.009] [PMID: 19358933]
[26]
Obeid, A.; Alawad, A.M.; Ibrahim, H.M. Isolation and characterization of bacillus subtillus with potential production of nattokinase. Int. J. Adv. Res., 2015, 3(3), 94-101.
[27]
Peng, Y.; Yang, X.; Zhang, Y. Microbial fibrinolytic enzymes: An overview of source, production, properties, and thrombolytic activity in vivo. Appl. Microbiol. Biotechnol., 2005, 69(2), 126-132.
[http://dx.doi.org/10.1007/s00253-005-0159-7] [PMID: 16211381]

Rights & Permissions Print Cite
© 2024 Bentham Science Publishers | Privacy Policy