A Statistical Application for the Enhanced Production of Streptokinase from a Mutant Strain Streptococcus equinus VIT_VB
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A Statistical Application for the Enhanced Production of Streptokinase from a Mutant Strain Streptococcus equinus VIT_VB2 Vaishnavi Babu1 • C. Subathra Devi1
Received: 14 June 2017 / Revised: 13 January 2020 / Accepted: 18 January 2020 The National Academy of Sciences, India 2020
Abstract The present study focuses on media optimization of streptokinase (SK) production from the mutant strain EMS1 of Streptococcus equinus VIT_VB2 which was originally isolated from bovine milk. A two-level Plackett– Burman design was applied to screen key ingredients of the media (fructose, tryptose, KH2PO4, MgCl2, NaHCO3, CH3COONa, temperature, pH and incubation time). The dependent variables influencing SK production was further carried out using central composite design using response surface methodology. Statistical analysis revealed that the second-order model was found to be significant with R2 value of 0.9895. The production of SK was found to be high with supplementation of fructose, tryptose and potassium dihydrogen phosphate. The mutant strain EMS1 gave 2.06-fold higher SK production in the presence of optimized medium (663.53 U mL-1) than that of parent strain (wild strain) in minimal basal medium (321.5 U mL-1). The optimization of process parameters using response surface methodology resulted in a twofold increase in the yield of fibrinolytic enzyme. Keywords Streptococcus equinus VIT_VB2 Streptokinase (SK) Plackett–Burman design Central composite design (CCD) Response surface methodology (RSM)
& C. Subathra Devi [email protected] Vaishnavi Babu [email protected] 1
Department of Biotechnology, School of Biosciences and Technology, VIT University, Vellore, Tamil Nadu 632 014, India
Approximately, 30% of mortality worldwide, 7.4 million deaths, were due to coronary heart disease, and 6.7 million deaths were due to stroke [1]. The knowledge about these diseases is abundant as these spectra of treatments are accessible in the form of clot-dissolving medication to surgical intervention. The primary assistance for the treatment of cardiovascular disorders (CVD) was aided by using clot dissolving or thrombolytic drugs. SK, one of the major plasminogen activators, is produced by different strains of b-haemolytic Streptococci. It is a protein, not an enzyme, and it combines with plasminogen to form the active complex that converts the free plasminogen to proteolysis plasmin. Hence, this characteristic feature of the protein has been extensively used in the treatment of thrombosis disorders, particularly acute myocardial infarction. The cost of SK has been reduced by 65% within short period of time due to increase in availability and easy affordable access. In addition, the use of media optimization and recombinant technology in large-scale production using bioreactor was developed by Council of Scientific and Industrial Research—Institute of Microbial Technology [2]. Statistical methodology was applied to optimize and fit the model using different variables such as carbon and nitro
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