Related Experiment Video
Updated: Jun 29, 2026
Using the Overlay Assay to Qualitatively Measure Bacterial Production of and Sensitivity to Pneumococcal Bacteriocins
Published on: September 30, 2014
Penicillium citrinum CFAM 521 Isolated From the Amazon Region: A Novel Source of a Fibrinolytic Enzyme
Thayana Cruz de Souza1, Marcos Gustavo Araujo Schwarz2, Daniela Marinho da Silva1
1Leônidas and Maria Deane Institute, ILMD/Fiocruz, Rua Teresina, 476, Adrianópolis, Manaus, Amazonas 69057-070, Brazil.
Abstract:
Fibrinolytic agents are essential in treating thrombosis, playing a critical role in improving survival rates in cardiovascular diseases. Microbial fibrinolytic proteases have emerged as promising alternatives due to their affordability, specificity, lower toxicity, and reduced side effects. Consequently, the search for microorganisms capable of producing these enzymes has gained significant economic importance in the pharmaceutical industry. This study reports and characterizes a novel fibrinolytic enzyme produced by Penicillium citrinum CFAM 521, a strain isolated from the Amazon region. The enzyme was purified using a polyethylene glycol (PEG)-phosphate salt aqueous two-phase system (ATPS). The effects of PEG molecular weight, PEG concentration, and phosphate concentration on the protease partition coefficient (K) were evaluated through a 22 full factorial design. The enzyme exhibited both fibrinolytic and fibrinogenolytic activities. After partitioning in a two-phase system with 10% (w/w) PEG and 15% (w/w) sodium phosphate, the fibrinolytic proteases were predominantly retained in the salt-rich bottom phase (K = 0.33). The enzyme has a molecular weight of 34 kDa, with optimal pH and temperature at 9°C and 37°C, respectively. Inhibitory analysis confirmed that it is a serine protease, and its activity was enhanced by the addition of Mn2+. Notably, the enzyme exhibited no hemolytic activity. Therefore, P. citrinum CFAM 521 represents a novel source of fibrinolytic enzymes, highlighting its potential as an alternative for the development of thrombolytic agents.
Insights
A novel fibrinolytic enzyme from Penicillium citrinum CFAM 521, isolated in the Amazon, shows potential as a safe and effective thrombolytic agent. This microbial protease offers a promising alternative to traditional treatments for thrombosis.
Area of Science:
- Biochemistry
- Microbiology
- Enzymology
Background:
- Fibrinolytic agents are crucial for treating thrombosis and improving cardiovascular disease survival.
- Microbial fibrinolytic proteases offer advantages like lower toxicity and cost-effectiveness compared to conventional agents.
- Identifying novel microbial sources for these enzymes is economically significant for the pharmaceutical industry.
Purpose of the Study:
- To report and characterize a novel fibrinolytic enzyme from Penicillium citrinum CFAM 521.
- To evaluate the enzyme's purification and properties for potential thrombolytic applications.
Main Methods:
- Isolation and cultivation of Penicillium citrinum CFAM 521 from the Amazon region.
- Purification of the fibrinolytic enzyme using a polyethylene glycol (PEG)-phosphate salt aqueous two-phase system (ATPS).
- Characterization of enzyme activity, including fibrinolytic, fibrinogenolytic, pH, temperature optima, molecular weight, and inhibition assays.
Main Results:
- A novel fibrinolytic enzyme was successfully purified from P. citrinum CFAM 521.
- The enzyme demonstrated both fibrinolytic and fibrinogenolytic activities, with optimal purification in a specific PEG-phosphate ATPS.
- Characterization revealed a 34 kDa serine protease, active at pH 9 and 37°C, enhanced by Mn2+, and notably lacking hemolytic activity.
Conclusions:
- Penicillium citrinum CFAM 521 is a novel source of fibrinolytic enzymes.
- The characterized enzyme exhibits properties suitable for potential development into thrombolytic agents.
- This microbial protease represents a promising, safe alternative for treating thrombosis.
Related Concept Videos
01:27Production of Antibiotics
01:15Antifungal Agents