Optimization of culture media for extracellular expression of streptokinase in Escherichia coli using response surface methodology in combination with Plackett-Burman Design

Aghaeepoor, Mojtaba; Kobarfard, Farzad; Eidgahi, Mohammad Reza Akbari; Shabani, Aliakbar; Dehnavi, Ehsan; Ahmadi, Maryam; Akbarzadeh, Ali

doi:10.4314/tjpr.v16i11.1

Cited by 8 publications

(6 citation statements)

References 24 publications

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“…Attaining an optimum expression state for a recombinant protein could be in uenced by different factors (11,12). It should be taken into account that the expression of a plasmid in the host cell brings to bear a metabolic burden which could terminate with diminished speci c growth rate and biomass content and plasmid instability (13). Various bacterial species do not have similar codon preferences for translation.…”

Section: Discussionmentioning

confidence: 99%

“…However, because of the high number of required experiments due to multiplex parameters, this approach is not feasible. In addition to the inconvenient nature of this approach, if there is an interaction between different variables, it can cause misinterpretation of the results (13,16). RSM is a popularly employed alternative procedure to overcome the aforesaid obstacles.…”

Section: Discussionmentioning

confidence: 99%

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Optimization of a recombinant BlaR-CTD protein formulation using the Response Surface Methodology

Abdolvahab,

Safari,

Hasannejad

et al. 2023

Preprint

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A penicillin-binding protein (PBP) is a group of proteins that have an affinity for and bind to penicillin. They are a normal component of many bacteria and play a crucial role in bacterial cell wall synthesis. PBPs are the targets of β-lactam antibiotics, such as penicillin. The sequence of a carboxy-terminal of the β-lactam sensor-transducer protein (BlaR-CTD) from Bacillus licheniformis ATCC14580 was extracted from US7745193B2 patent and expressed in E. coli using pColdI vector as a soluble His-tag recombinant protein. In this study, several excipients were used to improve the stability of recombinant BlaR-CTD and obtain the optimal formulation for this protein using response surface methodology (RSM)/ Central Composite Design (CCD). Total protein concentration was measured by UV spectroscopy and Bradford test. A total of 7 various formulations were designed using four different excipients including Glycerol, Sucrose, Triton x-100 and Tween-20, and three different buffers like Tris, Borate and PBS. By obtaining suitable excipients and buffer i.e. glycerol and sucrose, several concentrations of pH 7 to 9 were evaluated. The pH 7.62, glycerol 15.35% and sucrose 152.52 mM were determined as the most suitable for improve thermal stability of recombinant BlaR-CTD.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Optimization of a recombinant BlaR-CTD protein formulation using the Response Surface Methodology

Abdolvahab,

Safari,

Hasannejad

et al. 2023

Preprint

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show abstract

“…DoE is a statistical technique used to plan experiments and analyse data using a controlled set of tests designed to model and explore the relationship between factors and observed responses [14]. This technique allows the researcher to use the minimum number of experiments, in which the experimental parameters can be varied simultaneously, to make evidence based decisions [86]. It uses a mathematical model to analyse the process data, such as protein expression levels [87].…”

Section: Design Of Experiments (Doe) To Optimise Recombinant Protementioning

confidence: 99%

The Goldilocks Approach: A Review of Employing Design of Experiments in Prokaryotic Recombinant Protein Production

Uhoraningoga¹,

Kinsella²,

Henehan³

et al. 2018

Bioengineering

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The production of high yields of soluble recombinant protein is one of the main objectives of protein biotechnology. Several factors, such as expression system, vector, host, media composition and induction conditions can influence recombinant protein yield. Identifying the most important factors for optimum protein expression may involve significant investment of time and considerable cost. To address this problem, statistical models such as Design of Experiments (DoE) have been used to optimise recombinant protein production. This review examines the application of DoE in the production of recombinant proteins in prokaryotic expression systems with specific emphasis on media composition and culture conditions. The review examines the most commonly used DoE screening and optimisation designs. It provides examples of DoE applied to optimisation of media and culture conditions.

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“…Our research suggested that SK may be used clinically to treat cardiovascular disease. This technically advanced, stable, and valid animal model was created using ultrasonography and minimally invasive surgery, and it closely matched key clinical characteristics (Aghaeepoor et al, 2019). This could aid in improving our comprehension of thrombotic diseases and the creation of preventative and remedial measures.…”

Section: Introductionmentioning

confidence: 97%

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

Yousaf,

Arshad,

Harraz

et al. 2024

Braz. J. Biol.

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Cardiovascular disorders, including acute myocardial infarction (AMI), often lead to blood clot formation, impacting blood circulation. Streptokinase, a cost-effective and widely available thrombolytic agent, is crucial in treating thrombosis. This study aimed to produce streptokinase from Streptococcus pyogenes EBL-48 and compare its efficacy with heparin in an animal model. We evaluated the clot-lysing effectiveness of streptokinase produced from Streptococcus pyogenes EBL-48, emphasizing its low cost and ease of production. Streptokinase was produced using pre-optimized fermentation media and purified through ion exchange and gel-filtration chromatography. In vivo analysis involved inducing clots in a trial animal model using ferric chloride, comparing streptokinase with heparin. Ultrasonography assessed the clot-lysing activity of streptokinase. Streptokinase (47 kDa) effectively lysed clots, proving its low cost, easy production, and minimal adverse effects. Ultrasonography confirmed its fibrinolytic efficacy. These findings highlight potential as an affordable and easily produced thrombolytic agent, particularly relevant in resource-limited settings. Streptokinase efficacy and minimal adverse effects make it a promising option for thrombolytic therapy, especially in economically constrained regions. Future studies could optimize production techniques, explore different strains, and conduct clinical trials for human validation. Comparative studies with other thrombolytic agents would enhance understanding of their advantages and limitations.

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Optimization of culture media for extracellular expression of streptokinase in Escherichia coli using response surface methodology in combination with Plackett-Burman Design

Cited by 8 publications

References 24 publications

Optimization of a recombinant BlaR-CTD protein formulation using the Response Surface Methodology

Optimization of a recombinant BlaR-CTD protein formulation using the Response Surface Methodology

The Goldilocks Approach: A Review of Employing Design of Experiments in Prokaryotic Recombinant Protein Production

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

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