FIBRINOLYTIC ENZYME FROM BACILLUS AMYLOLIQUEFACIENS: OPTIMISATION AND SCALE UP STUDIES
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2014, Vol 6, Issue 10
Abstract
Objective: This research work was carried out to identify a potent microorganism, which produced the fibrinolytic enzyme and to optimise the media and growth parameters to achieve the maximal enzyme production for commercial application.Methods: Microorganisms were isolated from different sources and assayed for fibrinolytic activity. The shortlisted cultures after preliminary screening (casein hydrolysis, blood plate assay and blood clot dissolution) were identified using 16S rRNA amplification method. The media and growth parameters were optimized to achieve the maximal enzyme activity. In-silico studies were carried out to identify the activators and inhibitors of the enzyme.Results: Two species of Bacillus, namely, Bacillus amyloliquefaciens and Bacillus licheniformis, isolated from spoilt milk and soy flour, respectively, exhibited fibrinolytic activity. In the laboratory scale studies, of these two cultures, B. amyloliquefaciens produced the Fibrinolytic enzyme in higher quantities, 28.98 FU/mL, compared to 26.63 FU/mL in B. licheniformis. The maximal activities were obtained after 72 h. The optimum conditions at laboratory scale for the maximal production of the fibrinolytic enzyme were: pH 7.2, temperature 37 C and agitation 200 rpm. When scale up studies with B. amyloliquefaciens in a 7 L Fermentor were undertaken. The maximal activity obtained was 55.60 FU/mL in 72 h, compared to that of 28.98 FU/mL in shake flask studies. The molecular weight of the enzyme was estimated to be about 38 kDa. In in-silico studies, it was observed that PMSF inhibited the fibrinolytic activity, thereby, confirming this fibrinolytic enzyme is a serine protease (Nattokinase).Conclusion: The enzyme had exhibited excellent blood clot dissolving property and therefore may be considered for further scale up and commercial exploitation.Â
Authors and Affiliations
Rajani Gopal Gad, S Nirmala, S Narendar Sivvaswamy
Keywords
Related Articles
- EP ID EP579468
- DOI -
- Views 79
- Downloads 0
DRUG DISCOVERY OF NEWER ANALOGS OF ANTI-MICROBIALS THROUGH ENZYME-INHIBITION: A REVIEW
There is a growing interest towards the development of new antibiotics from last decades due to emergence of newer pathogenic bacterial strains with high resistance to powerful antibiotics of last resort. This has caused...
ESTIMATION OF STUNTED, WASTED, WASTED AND STUNTED CHILDREN AT TOWN DHADGAON, DISTRICT-NANDURBAR OF MAHARASHTRA STATE, INDIA
Objective: The aim of present study was to estimate the percentage of all three classes of malnutrition such as i) Stunted ii) wasted iii) wasted and stunted at study site, so that these findings could help to formulate...
A REVIEW ON CO-PROCESSED EXCIPIENTS: CURRENT AND FUTURE TREND OF EXCIPIENT TECHNOLOGY
There is no single-component excipient fulfills all the requisite performance to allow an active pharmaceutical ingredient to be formulated into a specific dosage form. Co-processed excipient has received much more atten...
MYRICETIN ISOLATED FROM TURBINARIA ORNATA AMELIORATES ROTENONE INDUCED PARKINSONISM IN DROSOPHILA MELANOGASTER
Objective: Parkinson’s disease (PD) is a neurodegenerative disorder which affects the elderly population. Free radicals overproduction, oxidative stress, apoptosis, inflammation and abnormalities in mitochondria are cr...
COMPARATIVE PHARMACOGNOSTICAL AND PHYTOCHEMICAL STUDY OF VARIOUS SPECIES OF THE CONTROVERSIAL DRUG ‘PUNARNAVA'
Objective: Punarnava is a controversial drug i.e. its true botanical identity has not been ascertained, and several different species are used as Punarnava. It is a part of many medicinally important traditional formulat...