The main aim of the review is to recapitulate the effects of thrombotic diseases, clotting disorders, and the methodology of prospecting a range of thrombolytics from various sources and thrombolytic therapies. Thrombolytic medications are administered to infringe dangerous clots inside blood vessels when there is deprivation in this haemostatic balance. Certain bioactive compounds present in herbs, plant leaves, roots and fruits exhibit thrombolytic activities. Serine proteases, metalloproteases and some cysteine proteases possess thrombolytic properties. These proteases are found in plants, humans, animals and microbes as well. Thrombolytics which are of clinical importance are discussed in this article along with its variants and biological sources exhibiting thrombolytic activity. Alternative therapies followed in recent days are also discussed, including future prospects of research on thrombolytics. Knowledge of thrombolytic biodiversity plays a decisive role in choosing safe, economic and effective thrombolytics for scale up and commercialization. © 2018, Institute of Korean Medicine, Kyung Hee University and Springer Science+Business Media B.V., part of Springer Nature.

Subathradevi C

Department of Biotechnology

School of Bio Sciences and Technology

Vellore Campus

Merlyn Keziah S

Vellore Institute of Technology (VIT) is a private university located in&nbsp;Tamil Nadu, India. Founded in 1984, as Vellore Engineering College, the institution offers 20 undergraduate, 34 postgraduate, four integrated and four research programs. It has campuses in Vellore, Amravati, Bhopal and Chennai.

VIT is one of the top ranked private universities in India according to NIRF, THE and QS Rankings.&nbsp;Govt. of India has recognized&nbsp;VIT, Vellore as an&nbsp;Institution of Eminence. This has allowed VIT to take independent quality initiatives and move up in world ranking.

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VIT University

Oriental Pharmacy and Experimental Medicine

Back ground: One of the most prevailing diseases that required effective drug as therapeutic purposes is cardiac related illness includes myocardial infarction Current scenario makes enzyme to raises their sector towards therapeutic as effective active component for such diseases. Objectives: The main aim of the study was to isolate, screen, characterize and produce an extracellular thrombolytic protease from marine actinomycetes. Methods: Marine actinomycete was isolated and characterized on the basis of morphological, biochemical, and molecular characterization. The primary screening for protease activity was done by casein hydrolysis method followed by radial caseinolytic assay. The actinoprotease was partially purified using ammonium sulfate precipitation technique followed by dialysis and Sephadex G-50 gel permeation chromatography. Results: 16srDNA sequencing and BLAST search analysis of the sequence revealed close affiliation with Streptomyces genera and identified as Streptomyces violaceus VITYGM with 99% similarity. The specific activity of purified protease was found to be 1437 units/mg along with purification fold up to 1.5 times. The blood clot lysis activity was compared with the standard and found to lyse the blood clot with 97.43%. Till now very less evidences have been reported on actinoprotease. A single peak at retention time 0.9 min observed on high performance liquid chromatography (HPLC) confirmed the homogeneity of the preparation. Conclusion: This is the first study to report on actinoprotease from Streptomyces violaceus VITYGM. This study emphasizes the potency of novel actinoproteases as active compound in drugs for the treatment of cardiovascular diseases. © 2016 Bentham Science Publishers.

Cardiovascular & Hematological Agents in Medicinal Chemistry

In vitro Thrombolytic Potential of Actinoprotease from Marine Streptomyces violaceus VITYGM

A potent and novel anticoagulant peptide was enzymatically isolated from the edible seaweed Porphyra yezoensis (commercially known as Nori). The pepsin hydrolysate showed 35% degree of hydrolysis and exhibited higher anticoagulant activity than undigested crude. The hydrolysate was fractionated by 4 sequential steps using size exclusion on Sephadex G-100, anion exchange, Sephadex G25 chromatography and desalting by Sephadex G-10. The purified peptide showed a single peak on RP-HPLC. The molecular mass of the peptide obtained by MS/MS analysis was 1.77 kDa and its sequence was identified as NMEKGSSSVVSSRM(+15.99)KQ. In vitro anticoagulation assays of purified peptide for clotting time with human plasma showed a dose-dependent prolongation of activated partial thromboplastin time from normal 35 s to 320 s with IC50 value of 0.3 μM. It did not prolong prothrombin or thrombin time. These results emphasise that the peptide interacts with the clotting factors involved in the intrinsic pathway of coagulation. © 2015 Elsevier Ltd.

Journal of Functional Foods

A novel anticoagulant peptide from the Nori hydrolysate

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. © 2015, Ahvaz Jundishapur University of Medical Sciences.

Fulltext

Jundishapur Journal of Microbiology

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

The aim of this study was to isolate and characterize streptokinase-producing β-hemolytic Streptococcus sp. from bovine milk. A total of 50 milk samples were collected randomly from different breeds of cow and goat (Vellore, Tamil Nadu, India). The samples were characterized and screened for streptokinase-producing isolates using microbial and biochemical analysis. About 97 colonies were isolated from milk samples showing hemolytic patterns of α (19.6%), β (24.7%) and γ (55.6%). Out of 20β-hemolytic isolates, only 6 colonies (VB2, VB3, VB8, VB14, VB16, and VB17) were identified as β-hemolytic Streptococci as potent producers of streptokinase. VB2 and VB14 showed the greatest streptokinase activities of 265 U mL-1 and 225 U mL-1, respectively. Based on biochemical and molecular characterization, the potent isolates VB2 and VB14 were identified and confirmed as S. equinus and S. agalactiae, respectively. The identified strains were named Streptococcus equinus VIT_VB2 (GenBank accession no. JX406835) and Streptococcus agalactiae VITVS5 (GenBank accession No. KF186620) The strains isolated from bovine milk provide a variance in the fibrinolytic activity on blood clots. The current study has demonstrated that the isolation of streptokinase producers from bovine milk, and the production of streptokinase from novel strain, enhanced the fibrinolytic activity. This study is the first to report that Streptococcus equinus produces streptokinase. © 2015 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation.

Journal	Data powered by TypesetOriental Pharmacy and Experimental Medicine
Publisher	Data powered by TypesetSpringer Science and Business Media LLC
ISSN	1598-2386
Open Access	No