Enhanced production of nattokinase from UV mutated Bacillus sp.

Mohanasrinivasan, V.; Devi, C. Subathra; Biswas, Ritusree; Paul, F. U. John; Ethiraj, Selvarajan; Suganthi, V.

doi:10.3329/bjp.v8i2.13690

Cited by 23 publications

(11 citation statements)

References 16 publications

(9 reference statements)

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“…The production of NK has been enhanced by physical and chemical mutagenesis. The results of the present study are supported by existing reports on enhanced production of NK by UV-mutated Bacillus spp., which significantly demonstrated better activity compared to the wild type strain ( 22 ).…”

Section: Discussionmentioning

confidence: 89%

“…In the Holmstrom method, the extent of lyses was checked after the coagulation of blood, in order to determine enzyme activity. One unit of enzyme activity was defined as the minimum amount of enzyme at which blood clot lysis occurred completely ( 18 , 22 , 23 ). Thereby, it can be concluded from the results that 60 and 90 minutes of exposure time to UV was the most effective in producing favorable mutation that enhanced NK production with a higher activity.…”

Section: Discussionmentioning

confidence: 99%

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Exploring the In Vitro Thrombolytic Activity of Nattokinase From a New Strain Pseudomonas aeruginosa CMSS

Чандрасекаран

Mohanasrinivasan

Shanker

et al. 2015

Jundishapur J Microbiol

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Background:Thrombolytic therapy has become a conventional treatment for acute myocardial infarction (AMI), yet currently, clinically prescribed thrombolytic drugs have problems such as delayed action and other side effects. Fibrinolytic enzymes have attracted interest as thrombolytic agents because of their efficiency in the fibrinolytic process, including plasmin activation. Nattokinase (NK) is a potent fibrinolytic agent for thrombosis therapy.Objectives:The aim of this study was to enhance the production of NK from Pseudomonas aeruginosa CMSS by media optimization and strain improvement.Materials and Methods:In the present study, a potent NK-producing strain was isolated from cow milk and identified. To enhance the yield of NK, effect of various parameters such as pH, temperature, carbon source, nitrogen source and inoculum size were optimized. Strain improvement of P. aeruginosa CMSS was done by random UV-mutagenesis. Nattokinase was partially purified and the activity was determined by the casein digestion method, blood clot lysis and fibrin degradation assay.Results:Based on morphological, biochemical and molecular characterization, the strain was confirmed as P. aeruginosa (GenBank accession number: JX112657), designated as P. aeruginosa CMSS. The optimum condition at pH 7 and temperature at 25˚C showed activity of NK as 1514 U mL-1 and 1532 U mL-1, respectively. Sucrose as the carbon source and shrimp shell powder (SSP) as the nitrogen source expressed NK activity of 1721 U mL-1 and 2524 U mL-1, respectively. At 1% inoculum size, the maximum rate of enzyme production was achieved with 2581 U mL-1. The NK activity of the mutant strain UV60 was 4263 U mL-1, indicating a two-fold increase in activity compared to the wild strain (2581 UmL-1). Nattokinase produced from mutant strain P. aeruginosa CMSS UV60 showed 94% blood clot lysis at ten minutes. The degradation of fibrin clot by the produced NK was observed after two hours of incubation. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the molecular mass of CMSS UV60 NK to be 21kDa.Conclusions:The current study demonstrated the enhanced production of NK by P. aeruginosa CMSS. This study is unique and the findings are the first report on the production of NK from P. aeruginosa CMSS isolated from cow milk.

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Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Exploring the In Vitro Thrombolytic Activity of Nattokinase From a New Strain Pseudomonas aeruginosa CMSS

Чандрасекаран

Mohanasrinivasan

Shanker

et al. 2015

Jundishapur J Microbiol

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“…Furthermore, P. aeruginosa CMSS grown on 1% sucrose supplemented medium, showed maximum NK activity (3930 U mL -1 ) than the strain grown in medium with other carbon source (Fig.3 d). The results of the present study are supported by existing reports on enhanced production of NK by UV-mutated Bacillus spp., which significantly demonstrated better activity compared to the wild type strain 13 . The previous studies on NK production after optimization reported that the maximum NK activity from Bacillus sp.…”

Section: Optimization Studiesmentioning

confidence: 90%

Strain improvement, production and stability of Nattokinase from UV mutant strain of Pseudomonas aeruginosa CMSS

Srivatsava¹,

Mohanasrinivasan²,

Naine³

et al. 2018

IJCTR

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Nattokinase (NK) is a potent fibrinolytic enzyme , which belongs to the second large family of serine proteases. NK gain significant attention in the treatment of hypertension and cardiovascular disorders (CVD's). Thus, a number of NK producers have been extensively studied, especially Pseudomonas sp. has extended the production level than Bacillus sp. with different properties. Thus the current study precedes with the strain improvement of NK producing UV mutated Pseudomonas aeruginosa CMSS by chemical mutagenesis. The potent mutant strain UV-EMS, treated with Ethyl Methyl Sulphonate (EMS) showed maximum NK activity. The maximum production of NK from mutant strain was determined at optimized parameters like pH 5 (1315.8 U mL-1), temperature at 37 o C (2413.3 U mL-1), shrimp shell as nitrogen source (2355.0 U mL-1) and sucrose as carbon source (3930.0 U mL-1).The activity of partially purified NK was stable at pH 7, temperature at 10 o C and 5mM of MgCl 2. The stability of NK was partially inhibited by SDS and completely inhibited by EDTA. The partially purified NK showed74% of in vitro clot lysis activity.

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“…The heated plasma was then cooled to optimum temperature and added to nutrient agar and mixed well by shaking (15ml of nutrient agar medium was prepared, 10 ml of plasma was added) and poured into petri dishes. Well diffusion was also used in this technique [17]. [18,19] with minor modification.…”

Section: -Human-heated Plasma Agar Testmentioning

confidence: 99%

Detection of Sak Gene and Expression of Staphylokinase in Different Clinical Isolates of Staphyloccocus spp

Abed¹,

AL-Azzawie²

2018

Tikrit J. Pure Sci.

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This study had investigated the presence of Staphylokinase (SAK) phenotypically and genetically. A total of 100 Staphylococcus spp. samples were isolated from different clinical cases. Morphological, biochemical tests and for further conformation; Viteck2 technique were adopted to isolate and diagnose the staphylococcal samples. Phenotypic assay tests including hydrolysis of casein and plasma agar were performed to check for the production of Staphylokinase from producing Staphylococcus spp. Out of 100, only 32 isolates had exhibited (SAK) activity whereby 29 of them represented the species S. aureus while 3 belonged to others. All isolates were subjected to Polymerase Chain Reaction (PCR) to determine the possessed sak gene. This investigation certainly had shown that not all isolates of S. aureus has the sak gene, only the lysogenic one has it and that it is not species committed but also other staphylococcus species could have it.

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Enhanced production of nattokinase from UV mutated Bacillus sp.

Cited by 23 publications

References 16 publications

Exploring the In Vitro Thrombolytic Activity of Nattokinase From a New Strain Pseudomonas aeruginosa CMSS

Exploring the In Vitro Thrombolytic Activity of Nattokinase From a New Strain Pseudomonas aeruginosa CMSS

Strain improvement, production and stability of Nattokinase from UV mutant strain of Pseudomonas aeruginosa CMSS

Detection of Sak Gene and Expression of Staphylokinase in Different Clinical Isolates of Staphyloccocus spp

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