scholarly journals Investigation of isolation conditions and ion-exchange purification of protein coagulation components from common bean seed

2007 ◽  
pp. 3-10
Author(s):  
Mirjana Antov ◽  
Marina Sciban ◽  
Slavica Adamovic ◽  
Mile Klasnja
Keyword(s):  
Ion Exchange ◽  
Common Bean ◽  
Crude Extract ◽  
Protein Concentration ◽  
Extraction Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Bean Seed ◽  
Coagulation Activity ◽  
Water As Solvent

Investigation of an extraction procedure of protein coagulants from common bean seed regarding concentration of NaCl and pH was performed. High values of protein concentration and coagulation activity in crude extract (9.19 g/l and 23.9%, respectively) were obtained when the extraction was performed using 0.5 mol/l NaCl and water as solvent, which represents an advantage for economic and environmental reasons. Crude extract of common bean seed was purified by precipitation at two different percentages of (NH4)2SO4 saturation, followed by batch ion-exchange chromatography. The highest obtained coagulation activity, 45%, was determined in fraction that was eluated at 1.75 mol/l NaCl from resin loaded with proteins precipitated upon 80-100% (NH4)2SO4 saturation. High values of coagulation activity showed by some eluates suggest their application as natural coagulant for water purification. .

scholarly journals Control of 5-aminolaevulinate synthetase activity in Rhodopseudomonas spheroides a role for trisulphides

1975 ◽  
Vol 150 (2) ◽  
pp. 245-257 ◽  
Author(s):  
J D Sandy ◽  
R C Davies ◽  
A Neuberger
Keyword(s):  
Ion Exchange ◽  
Extraction Procedure ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Synthetase Activity ◽  
Multiple Forms ◽  
Anaerobic Culture ◽  
Structural Requirement ◽  
Rhodopseudomonas Spheroides ◽  
Low Activity

1. The aminolaevulinate synthetase activator of fresh extracts of semi-anaerobically grown Rhodopseudomonas spheroids was resolved into two fractions by ion-exchange chromatography. One fraction was identified as cystine trisulphide (CySSSCy). Analysis of the other fraction indicated the presence of about equal amounts of glutathione trisulphide (GSSSG) and the mixed trisulphide of glutathione and cystine (GSSSCy). 2. Four further fractions with activator activity were observed on ion-exchange chromatography of extracts prepared by methods similar to those described earlier [Neuberger et al. (1973)Biochem. J. 136,491-499]. These activators were generated by the extraction procedure. Two of them have been identified as trisulphanedisulphonate (S5O62-) and additional cystine trisulphide. 3. For the series of polysulphanedisulphonates (-O3S-Sn-SO3-, n greater than or equal to 1), activator activity at muM concentrations was exhibited only by compounds with n greater than 3. This, together with the results described above, indicates that for a compound R-Sn-R' (where R and R' are organic or inorganic groups) the only structural requirement for activity is n greater than or equal to 3. 4. Oxygenation of a semipanaerobic culture of R. spheroids for 1.5h before harvesting the cells produced a decrease of more than 90% in the cellular content of cystine trisulphide and glutathione trisulphides. 5. Chromatography on DEAE-Sephadex confirmed the presence of multiple forms of aminolaevulinate synthetase in extracts of R. spheroides [Tuboi et al. (1970) Arch. Biochem. Biophys. 138,147-154]. Oxygenation of a semi-anaerobic culture resulted in the disappearance of high-activity enzyme (a-form) and the accumulation of low-activity enzyme (b-form) in the cell. Spontaneous activation [Marriott et al. (1969) Biochem. J. 111,385-394] And activation by cystine trisulphide both resulted in the almost complete conversion of the b-form into the a-form.

scholarly journals Ultrafiltration as a simple purification method of a water extract of common bean seed as a natural coagulant

2020 ◽  
Vol 74 (3) ◽  
pp. 211-220
Author(s):  
Jelena Prodanovic ◽  
Bojana Saric ◽  
Marina Sciban ◽  
Dragana Kukic ◽  
Vesna Vasic ◽  
...  
Keyword(s):  
Organic Matter ◽  
Common Bean ◽  
Crude Extract ◽  
Water Extract ◽  
Bean Seed ◽  
Coagulation Activity ◽  
Turbidity Removal ◽  
Molecular Weights ◽  
Natural Coagulants ◽  
Model Water

Natural coagulants from a crude water extract of common bean seed showed very good efficiency of turbidity removal from water of ~89 % under optimal coagulation conditions, which were determined using response surface methodology (RSM). However, they also increased the content of organic matter in treated model water by ~66 %, which is the main drawback of usage of natural coagulants, in general. Thus, ultrafiltration was applied for processing of the crude water extract in order to separate biomolecules, which exhibit the coagulation activity. Four fractions obtained by ultrafiltration were applied in coagulation tests under the same conditions as the crude extract, and the 4th fraction (molecules with molecular weights >30 kDa) with the predominant content of proteins with molecular weights 50 - 60 kDa, achieved almost as high efficiency of turbidity removal (75 %) as the crude extract. At the same time, the content of organic matter in treated water increased just for 16 % in comparison to the blank (model water processed in the same way but without coagulant). After optimization of process parameters by RSM for usage of the 4th fraction, the coagulation activity increased further to 80 %.

scholarly journals Extraction and partial purification of coagulation active components from common bean seed

2006 ◽  
pp. 37-43 ◽  
Author(s):  
Marina Sciban ◽  
Mirjana Antov ◽  
Mile Klasnja
Keyword(s):  
Common Bean ◽  
Anion Exchange ◽  
Crude Extract ◽  
Partial Purification ◽  
Turbid Water ◽  
Nacl Solution ◽  
Bean Seed ◽  
Active Components ◽  
Coagulation Activity ◽  
Nacl Concentration

An active coagulation component was extracted from common bean seed by NaCl solution and the obtained crude extract was partially purified through a sequence of steps that included precipitation of protein by ammonium sulphate, desalting by dialysis and anion exchange. A turbid water was treated by protein fractions obtained in the anion- exchange elution process by stepwise increase in NaCl concentration. The jar tests were conducted at various dosages of eluates. Different mode of relation between coagulation activity and applied coagulant dose for each protein fraction indicated the existence of different mechanisms of coagulation/flocculation, depending of characteristics of different proteins in the fractions.

scholarly journals Interaction between S-Type Pyocins and Microcin-II-Like Bacteriocins in Pseudomonas aeruginosa

2021 ◽  
Vol 83 (3) ◽  
pp. 72-80
Author(s):  
O.B. Balko ◽  
Keyword(s):  
Molecular Weight ◽  
Pseudomonas Aeruginosa ◽  
Ion Exchange ◽  
Protein Concentration ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Activity Increase ◽  
Activity Indices

According to our previous results, S-type bacteriocins of Pseudomonas aeruginosa are characterized by high activity against phytopathogenic Pseudomonas syringae strains. In addition to these pyocins producing strains are able to synthesize microcin-II-like bacteriocins. Presence of interaction between these two killer factors can determine methods of their use and activity increase of bacteriocins with antiphytopathogenic properties. The aim of the work was to test possibility of interaction between S-type pyocins and microcin-II-like bacteriocins of P. aeruginosa. Methods. The objects of the study were pyocins produced by 6 P. aeruginosa strains. Killer factors in composition of induced lysates were concentrated by 70% ammonium sulphate precipitation, dialyzed through dialysis membrane with molecular weight cut-off (MWCO) 3.5 kDa. Then ion-exchange chromatography with DEAE-cellulose, gel filtration with Sephadex G-75 and ultracentrifugation at 215.000 g for 1 and 4 hours were used for their separation. Protein concentration and antimicrobial activity were determined in obtained fractions. Visualization of proteins in active fraction composition was conducted by electrophoresis according to the Laemmli method. Results. Under ion-exchange chromatography with DEAE-cellulose application elution of bacteriocins available in lysate composition occurs simultaneously. The highest indices of activity and protein concentration were in the 4th fraction, containing two protein bands with molecular weight near 58 and 9 kDa, which are typical for S5 pyocin and microcin-II-like bacteriocins of P. aeruginosa. Further gel filtration of sampled fractions through Sephadex G-75 allowed to separate noted killer factors and obtaine purified fraction containing microcin-II-like pyocins only. Application of ultracentrifugation during 1 hour didn’t precipitate studied bacteriocins, whereas during 4 hours – lead to their separation. At the same time a twofold increase of activity indices for S-type pyocins in precipitates and for microcin-IIlike killer factors – in supernatants were observed. However achieved concentration was characterized by short-term effect, since in 14 days activity of supernatants decreased by 4–16 times, and for precipitates – by 80–640 times. Then revealed tendency for activity decrease continued. Conclusions. S-type pyocins and microcin-II-like bacteriocins of P. aeruginosa interact with each other, that ensures their stabilization and protects again destruction. Application of methods that cause separation of these killer factors is inexpedient, since it results into considerable decrease of bacteriocin activity indices.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

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