scholarly journals PRODUCTION AND PARTIAL CHARACTERIZATION OF FIBRINOLYTIC ENZYME FROM A SOIL ISOLATE ASPERGILLUS CARBONARIUS S-CSR-0007

2016 ◽  
Vol 8 (12) ◽  
pp. 142 ◽  
Author(s):  
Afini A.v. M. ◽  
Sooraj S. Nath ◽  
Smitha K. V. ◽  
Kunhi A.a. M.
Keyword(s):  
Ammonium Sulfate ◽  
Substrate Concentration ◽  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Maximum Activity ◽  
Fibrinolytic Enzyme ◽  
Clot Lysis ◽  
Aspergillus Carbonarius ◽  
Lysis Activity ◽  
Wide Range

<p><strong>Objective: </strong>This work was undertaken with the aim of isolating and screening fungal soil isolates with fibrinolytic activity.</p><p><strong>Methods: </strong>Soil sample near slaughter house was collected and screened for fibrinolytic activity by using fibrin-agar. Enzyme production was optimized under various parameters like pH, temperature, substrate concentration and purified partially by ammonium sulphate precipitation. The stability of the partially purified enzyme was analyzed under the influence of a wide range of pH, temperature, and substrate concentrations.<strong></strong></p><p><strong>Results: </strong>Among the seven isolates screened, <em>Aspergillus carbonarius</em> S-CSR-0007 exhibited largest clear zone and was selected for further studies. Among the various substrates tested casein was found to support the highest caseinolytic activity of 816 U/ml and fibrinolytic activity of 510 U/ml. The culture supernatant of <em>A. carbonarius</em> S-CSR-0007 was fractionated by ammonium sulfate precipitation followed by dialysis, and maximum activity was obtained in the fraction with 80% ammonium sulfate, with an enzyme activity of 1200 U/ml using tyrosine as standard. The partially purified fibrinolytic enzyme showed optimal activity at 45 °C and pH 7.0. The enzyme was stable up to a temperature of 50 °C and pH 8.0, and the optimum substrate concentration was 4%.</p><p><strong>Conclusion: </strong>The crude enzyme showed high blood clot lysis activity, which may be a good candidate in the pharmaceutical industry. However, more studies need to be carried out to establish its clinical use.</p>

Production of fibrinolytic protease from a halobacterium Bacillus licheniformis VITLMS isolated from marine sponges of Rameshwaram coast, India

2020 ◽  
Vol 16 ◽  
Author(s):  
Lalith Kumar .CK ◽  
Merlyn Keziah.S ◽  
Hemalatha. M ◽  
Mohanapriya.A ◽  
Mohanasrinivasan.V ◽  
...  
Keyword(s):  
Ammonium Sulfate ◽  
Bacillus Licheniformis ◽  
Fibrinolytic Activity ◽  
Gel Filtration ◽  
Marine Sponges ◽  
Clot Lysis ◽  
Malt Extract ◽  
Ammonium Sulfate Precipitation ◽  
Fibrinolytic Enzymes ◽  
Lysis Activity

Background: Marine bacteria serve as excellent sources of therapeutic enzymes, metabolites and natural products, which possess novel therapeutic properties. Increasing death rates due to cardiovascular diseases urges for cost effective production of fibrinolytic enzyme. Methods: Marine sponge samples were screened for potent fibrinolytic producing bacteria. Primary screening was done for protease production, clot lysis activity. Secondary screening was done for casein plasminogen activity and fibrinolytic activity. The strain which had potent fibrinolytic activity among them was further subjected to morphological, biochemical and molecular characterization. Media optimization was carried to enhance enzyme production. The enzyme produced was subjected to purification using ammonium sulfate precipitation, gel filtration and characterized using HPLC and FTIR analysis Results: Sponge was identified to be Desmapsamma anchorata. Thirteen bacterial isolates were isolated from the sponge sample. . The 16S rRNA sequencing revealed that the potential strain had 99% similarity with Bacillus licheniformis. Amongst the isolates most were found to be morphologically identical to Bacillus genus. Gram’s staining and SEM analysis of the potent isolate was performed to identify the spore formation and rod shaped morphology of the bacteria. The optimal temperature and pH for production of the enzyme was 37 °C and 8 respectively. The carbon source maltose and nitrogen sources were malt extract and yeast extract were found to be optimal. The optimum incubation time was found to be both 4 to 5 days. The crude supernatant was purified with ammonium sulfate precipitation and gel filtration chromatography. The retention time of 11.3 min and presence of functional groups show the purity of the enzyme. The partially purified enzyme showed 96.4 % clot lysis in artificial clot lysis activity. Conclusion: Although the secretion of fibrinolytic enzymes from Bacillus species is not new, based on our investigation there are no reports regarding Bacillus licheniformis being isolated from marine sponges. However, there are reports of Bacillus licheniformis secreting fibrinolytic enzymes isolated from fermented food samples This study identifies marine environment as a potential source of new exploration for drug discovery.

scholarly journals Novel Fibrinolytic Protease Producing Streptomyces radiopugnans VITSD8 from Marine Sponges

Marine Drugs ◽  
2019 ◽  
Vol 17 (3) ◽  
pp. 164 ◽  
Author(s):  
Dhamodharan D ◽  
Jemimah S ◽  
Merlyn S ◽  
Subathra C
Keyword(s):  
Tamil Nadu ◽  
Specific Activity ◽  
Blood Clot ◽  
Exchange Chromatography ◽  
Marine Sponges ◽  
Protease Production ◽  
Clot Lysis ◽  
Carbon And Nitrogen ◽  
Lysis Activity

Fibrinolytic enzymes have received more attention due to their medicinal potential for thrombolytic diseases. The aim of this study is to characterize the in vitro fibrinolytic nature of purified protease producing Streptomyces radiopugnans VITSD8 from marine brown tube sponges Agelas conifera. Three varieties of sponge were collected from the Rameshwaram Sea coast, Tamil Nadu, India. The fibrinolytic activity of Streptomyces sp. was screened and determined by casein plasminogen plate and fibrin plate methods respectively. The crude caseinolytic protease was purified using ammonium sulfate fractionation, affinity and ion-exchange chromatography. Based on the morphological, biochemical, and molecular characterization, the isolate VITSD8 was confirmed as Streptomyces radiopugnans. Maltose and peptone were found to be the best carbon and nitrogen sources for the production of fibrinolytic protease. The carbon and nitrogen source peptone showed (781 U/mL) enzyme activity. The optimum pH and temperature for fibrinolytic protease production was found to be 7.0 and 33 °C respectively. The purified enzyme showed a maximum specific activity of 3891 U. The blood clot lysis activity was compared with the standard, and it was concluded that a minimum of 0.18 U (10 µL) of purified protease was required to dissolve the blood clot. This is the first report which exploits the fibrinolytic protease activity of Streptomyces radiopugnans VITSD8 extracted from a marine sponge. Hence the investigation suggests a potential benefit of purified fibrinolytic protease which will serve as an excellent clot buster alternative.

Changes with age in Fibrinolytic Activity, Coagulation Factors and Lipids in an Industrial Population

1975 ◽  
Author(s):  
R. Chakrabarti ◽  
M. Brozovic ◽  
T. W. Meade ◽  
W. R. S. North ◽  
Y. Stirling
Keyword(s):  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Coagulation Factors ◽  
Blood Cholesterol ◽  
Factor Vii ◽  
Clot Lysis ◽  
Factor V ◽  
Cholesterol Levels ◽  
Lysis Time ◽  
Similar Factor

Fibrinolytic activity, coagulation factors I, V, VII and VIII, blood cholesterol and triglyceride levels have been measured in a randomyl selected group of 650 men aged 17–64 and 350 women aged 17–59 working in an industrial population. Fibrinolytic activity (the reciprocal of the dilute blood clot lysis time in hours) decreases significantly (0.003 per annum) with advancing age in men; there is no significant change in women. Factors I and V increase significantly with age in both men and women at about the same rate (1.0% per annum for factor I and 0.6% for factor V), their mean levels in each group being very similar. Factor VIII also increases significantly with age in both sexes up to the age of about 50 years, after which levels continue to rise in men, but fall in women. Factor VII and blood cholesterol levels are lower in young women than in young men; they rise in both sexes, but significantly faster in women than in men (1.1% and 0.4%, respectively for factor VII, for example), so that in older women they are substantially higher than in older men. Triglyceride levels rise with age in both sexes, levels in men being hig er than those in women.

ENDOTOXIN INDUCES DIC BY BOTH TISSUE FACTOR (TF) AND PLASMINOGEN ACTIVATOR-INHIBITOR (PA-l) SECRETION

1987 ◽  
Author(s):  
B Baldus ◽  
G Gehrmann ◽  
W Witt ◽  
P Donner
Keyword(s):  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Plasminogen Activator Inhibitor ◽  
Clot Lysis ◽  
E Coli ◽  
Microtiter Plates ◽  
Lysis Time ◽  
Male Wistar Rats ◽  
Baseline Values ◽  
Activator Inhibitor

DIC-induced organ failure represents a major pathomecha-nism in endotoxemia. To investigate which components of the haemostatic system may be disturbed during endotoxemia anaesthetized male Wistar rats were injected with E. coli endotoxin at dosages of 1 ng/kg to 100 pg/kg body mass. Citrated blood samples were investigated for fibrinolytic activity by a dilute blood clot-lysis time assay (DBC-LT), for PA-I activity by a new Tunctional assay using immobilized rt-PA on 96-well microtiter plates and for TF activity by measuring the rate of thrombin formation after stimulation with a 2000-fold diluted thrombo plastin reagent.Endotoxin induced a decrease in fibrinolytic activity measured as a prolongation of the DBC-LT at dosages from 75 ng/kg to 100 μg/kg. This effect appeared 2 h after the treatment with endotoxin and persisted for<3 h. In parallel, an increase (up to 7-fold compared to baseline values) in PA-I activity could be measured 2 h after endotoxin application. When the plasma samples were clotted either by kaolin-phospholipid reagent, thrombin or reptilase in the resulting serum PA-I activity was more than 50-fold versus baseline. TF activity generated after stimulation with diluted thromboplastin reagent increased to about 5-fold compared to baseline values. This effect was already significant 1 h after endotoxin injection.The results of this study indicate, that endotoxin induces a destruction of the balance of the haemostatic system by increasing coagulability via stimulation of TF activity and by decreasing fibrinolytic activity by secretion of PA-I. Furthermore the present data provide evidence for activation of a latent form of PA-I during the clotting process.

scholarly journals The pathogenesis of accelerated fibrinolysis in liver cirrhosis: a critical role for tissue plasminogen activator inhibitor

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1315-1319 ◽  
Author(s):  
SL Hersch ◽  
T Kunelis ◽  
RB Jr Francis
Keyword(s):  
Liver Cirrhosis ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Critical Role ◽  
Plasminogen Activator Inhibitor ◽  
Clot Lysis ◽  
Lysis Time ◽  
Tissue Plasminogen

Abstract The pathogenesis of accelerated fibrinolysis in liver cirrhosis was investigated by comparing the results of specific assays for tissue plasminogen activator (tpa) antigen, tpa activity, tpa inhibitor, and alpha-2 plasmin inhibitor (a2PI) in 12 patients with cirrhosis and markedly accelerated fibrinolysis (dilute whole blood clot lysis time (DWBCLT) less than two hours), in nine patients with cirrhosis and moderately accelerated fibrinolysis (DWBCLT two to four hours), and in nine patients with cirrhosis and normal fibrinolysis (DWBCLT greater than four hours). Mean tpa antigen was markedly increased in all three groups, but no correlation was observed between overall fibrinolytic activity as measured by the DWBCLT and the level of tpa antigen. In contrast, there was a significant correlation between overall fibrinolytic activity and tpa activity and an equally significant correlation between fibrinolytic activity and decreased tpa inhibition. Mean a2PI activity was significantly lower than normal in groups 1 and 2 but was normal in group 3. The pathogenesis of accelerated fibrinolysis in liver cirrhosis thus appears to depend critically on the capacity of plasma inhibitors to inhibit increased circulating tpa antigen. Reduced a2PI also appears to play a role.

Adriamycin (A) Induces a Delayed Hypercoagulable and Prothrombotic State in Rats.

1979 ◽  
Author(s):  
F. Delaini ◽  
A. Popgi ◽  
T. Colombo ◽  
L. Komblihtt ◽  
I. Reyers ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Fibrinolytic Activity ◽  
Blood Clot ◽  
Clot Lysis ◽  
Limited Attention ◽  
Prothrombotic State ◽  
Occlusion Time ◽  
Protein Levels ◽  
Acute Changes

The effects of anticancer agents. on the host’s haemostatic mechanism have been given limited attention so far. We have studied the early and long-term changes in haemostatic parameters of normal rats given A. Single doses of A (5-10 mg/kg b.w.), at levels active as an anticancer agent in this animal species, did not induce any acute changes in platelets, fibrinogen or fibrinolytic activity, as long as plasma drug levels were measurable. Two weeks later, however, a hypercoagulable stat developed, characterized by increased fibrinogen and F.VIII:C activity, short APTT, low total antithrombin and heparin cofactor levels and markedly depressed fibrinolytic activity (measured by euglobulin and dilute whole blood clot lysis times, plasminogen and antiplasmin amidolytic activity). These changes were dose-related and lasted at least five weeks after treatment. When the plasma antithrombin level was low, this inhibitor could be measured in the urine. A significant reduction in the occlusion time of an aortic loop was observed in A-treated rats. The occurrence of ascites, proteinuria and reduced plasma protein levels in the same animals suggests that A-induced nephrotoxicity (nephrotic syndrome?) could be involved in the pathogenesis of the hypercoagulable and prothrombotic state observed.Supported by Contract NIH-NCI-C-72-3242, NIH (USA) and Italian CNR Project “Control of Tumoral Growth”.

Thrombolytic Properties of a Tissue Activator of Plasminogen

1979 ◽  
Author(s):  
G.V. Andreenko ◽  
E.E. Shimonaeva ◽  
T.N. Serebryakova
Keyword(s):  
Intravenous Injection ◽  
Fibrinolytic Activity ◽  
Degradation Products ◽  
Blood Clot ◽  
Blood Stream ◽  
Rat Plasma ◽  
Physiological Solution ◽  
Clot Lysis ◽  
Lysis Time

Preparations of a tissue activator of plasminogen (TA) readily soluble in the physiological solution were obtained from porcine heart according to the procedure of Bachman et al. up to the stage of precipitation by Zn( C2H3O2)2. H2O. “In vitro” the TA preparations reduced the lysis time of the rat plasma euglobulin fraction clot from 65-90 min in the control down to 7-14 min and induced the blood clot lysis. The highest rate of lysis was observed when the activator was combined with plasminogen. When TA was dissolved in rat plasma and physiological solution, the rate of the clot lysis was decreased. An intravenous injection of TA into the blood stream of rats with experimental thrombosis led to a clot lysis for 15-60 min. The rate of clot lysis was independent of the dose used. After 10 and 20 min of intravenous injection of TA the fibrinolytic activity of the plasma euglobulin fraction was increased 1,5-2-fold. The level of fibrinogen tended to a decrease. The level of degradation products was increased 1,2-1,5-fold. The fibrinolytic activity was slightly increased after 20 min. The activities of antiplasmi ns and antiactivator and the recalcification time remained unchanged.

Cohparison of Variability of Different Methods of Measuring Fibrinolytic Activity

1979 ◽  
Author(s):  
R. Chakrabarti ◽  
S.G. Thompson ◽  
T.W. Meade
Keyword(s):  
Fibrinolytic Activity ◽  
Large Scale ◽  
Blood Clot ◽  
Clot Lysis ◽  
Laboratory Error ◽  
Lysis Time ◽  
Fibrin Plate ◽  
Heart Study ◽  
Euglobulin Lysis Time ◽  
Plate Area

The Northwick Park Heart Study has drawn attention to the variability, especially within-person, of the dilute blood clot lysis time (DBCLT). However, this method has considerable practical advantages in large-scale surveys. The variability of OBCLT has therefore been compared with that of the euglobulin. lysis time (ELT) and the euglobulin fibrin plate area (EFPA). Each measurement has been made 10 times over a 9 month period in each of 12 healthy people; fibrinogen levels have also been measured. As expected, highly significant correlations (r) have been obtained between the three methods: DBCLT and ELT, + 0.63 DBCLT and EFPA, - 0.47; ELT and EFPA, - 0.54 (P<0.001 in each case). The between-pernon variance, distinguishing one person from another, accounts for 60% of the total vnriance in DBCLT, 35% in ELT and 10% in SFPA. For fibrinogen, the value is nearly 90%. The rest of the variance in each case is accounted for by hiological within-person variation and Laboratory error. Thus, while DBCLT is a variable method of determining an individual’s characteristic level of fibrinolytic activity, it is probably lees so than other commonly used methods. The results of this experiment suggest that a previous estimate of the within-person variability of DBCLT (Meade and North, 1977, British Medical Bulletin, 33, 283) may have been too high.

Sign in / Sign up

Export Citation Format

Share Document