scholarly journals Lactococcin Q, a Novel Two-Peptide Bacteriocin Produced by Lactococcus lactis QU 4

2006 ◽  
Vol 72 (5) ◽  
pp. 3383-3389 ◽  
Author(s):  
Takeshi Zendo ◽  
Shoko Koga ◽  
Yasushi Shigeri ◽  
Jiro Nakayama ◽  
Kenji Sonomoto
Keyword(s):  
Antibacterial Activity ◽  
Lactococcus Lactis ◽  
Exchange Chromatography ◽  
Molar Ratio ◽  
Significant Activity ◽  
Cation Exchange Chromatography ◽  
Reverse Phase ◽  
Wide Range ◽  
Molecular Masses ◽  
The Difference

ABSTRACT A bacteriocin-producing strain, Lactococcus lactis QU 4, was isolated from corn. The bacteriocin, termed lactococcin Q, showed antibacterial activity only against L. lactis strains among a wide range of gram-positive indicator strains tested. Lactococcin Q was purified by acetone precipitation, cation exchange chromatography, and reverse-phase chromatography. Lactococcin Q consisted of two peptides, α and β, whose molecular masses were determined to be 4,260.43 Da and 4,018.36 Da, respectively. Amino acid and DNA sequencing analyses revealed that lactococcin Q was a novel two-peptide bacteriocin, homologous to lactococcin G. Comparative study using chemically synthesized lactococcin Q (Qα plus Qβ) and lactococcin G (Gα plus Gβ) clarified that hybrid combinations (Qα plus Gβ and Gα plus Qβ) as well as original combinations showed antibacterial activity, although each single peptide showed no significant activity. These four pairs of lactococcin peptides acted synergistically at a 1:1 molar ratio and exhibited identical antibacterial spectra but differed in MIC. The MIC of Qα plus Gβ was 32 times higher than that of Qα plus Qβ, suggesting that the difference in β peptides was important for the intensity of antibacterial activity.

scholarly journals Proteinaceous Protease Inhibitor from Lawsonia Inermis: Purification, Characterization and Antibacterial Activity

2013 ◽  
Vol 8 (10) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Arvind Dabhade ◽  
Priti Patel ◽  
Ulhas Patil
Keyword(s):  
Antibacterial Activity ◽  
Protease Inhibitor ◽  
Deae Cellulose ◽  
Gel Permeation Chromatography ◽  
Apparent Molecular Weight ◽  
Exchange Chromatography ◽  
Lawsonia Inermis ◽  
Sds Page ◽  
Wide Range ◽  
And Staphylococcus Aureus

A thermo-stable, proteinaceous protease inhibitor (LPI) from Lawsonia inermis is reported. The LPI was purified from Lawsonia inermis seeds by subsequent ammonium sulfate precipitation, ion exchange chromatography (DEAE-Cellulose) and gel permeation chromatography (Sephadex-50). The purified protease inhibitor is effective against a wide range of proteases viz. papain, trypsin, pepsin and metallo-protease. The apparent molecular weight of the protease inhibitor is 19 kDa, determined by SDS-PAGE electrophoresis. The protease inhibitor was found to be stable at 70 °C for 30 min. It was also examined for antibacterial activity against Pseudomonas aeruginosa MTCC 7926 and Staphylococcus aureus NCIM 2079; the IC50 values of the purified LPI were 11.4 μg/mL and 16.6 μg/mL respectively.

scholarly journals Biochemical and Functional Characterization ofParawixia bistriataSpider Venom with Potential Proteolytic and Larvicidal Activities

2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
Author(s):  
Gizeli S. Gimenez ◽  
Antonio Coutinho-Neto ◽  
Anderson M. Kayano ◽  
Rodrigo Simões-Silva ◽  
Frances Trindade ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Functional Characterization ◽  
Exchange Chromatography ◽  
Biotechnological Applications ◽  
Cation Exchange Chromatography ◽  
Crude Venom ◽  
Molecular Masses ◽  
Larvicidal Activities ◽  
Parawixia Bistriata ◽  
Chromatographic And Electrophoretic Profiles

Toxins purified from the venom of spiders have high potential to be studied pharmacologically and biochemically. These biomolecules may have biotechnological and therapeutic applications. This study aimed to evaluate the protein content ofParawixia bistriatavenom and functionally characterize its proteins that have potential for biotechnological applications. The crude venom showed no phospholipase, hemorrhagic, or anti-Leishmania activities attesting to low genotoxicity and discrete antifungal activity forC. albicans. However the following activities were observed: anticoagulation, edema, myotoxicity and proteolysis on casein, azo-collagen, and fibrinogen. The chromatographic and electrophoretic profiles of the proteins revealed a predominance of acidic, neutral, and polar proteins, highlighting the presence of proteins with high molecular masses. Five fractions were collected using cation exchange chromatography, with the P4 fraction standing out as that of the highest purity. All fractions showed proteolytic activity. The crude venom and fractions P1, P2, and P3 showed larvicidal effects onA. aegypti. Fraction P4 showed the presence of a possible metalloprotease (60 kDa) that has high proteolytic activity on azo-collagen and was inhibited by EDTA. The results presented in this study demonstrate the presence of proteins in the venom ofP. bistriatawith potential for biotechnological applications.

scholarly journals A Polysaccharide Produced by Lactococcus lactis subsp. lactis YZ1 Isolated from Traditional Indonesian Fermented Milk, "Dadih"

2000 ◽  
Vol 1 (1) ◽  
pp. 1-5
Author(s):  
Yusdar Zakaria
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Lactococcus Lactis ◽  
Monosaccharide Composition ◽  
Fermented Milk ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Molar Ratio ◽  
Neutral Sugar ◽  
Glycoside Linkage

ABSTRACT.Lactococcus lactis subsp. Lactis YZI was isolated from M17 agar in which diluted Dadih was poured and incubated at 30 0C for 48 h. Taxonomix properties of the isolate were examined according to Bergey’s Manual of Systematic Bacteriologi and Manual for  Identification of Medical Bacteria. The isolation of polysaccharide from the precipitant was performed on an ion-exchange chromatography. The result showed that the polysaccharides produced by Lactococus lactis subsp. lactis YZI were neutral sugar (unadsorbrd fraction) and glycoconjugated (absorbed fraction). The neutral sugar had molecular weight of 10,000 and 20,000 with and α-glycoside linkage. The monosaccharide composition was mannose, glucose and galactose with a molar ratio of 1 :1,5 : 4,9.

Quantification of urinary oxalate with oxalate oxidase from beet stems.

1983 ◽  
Vol 29 (10) ◽  
pp. 1815-1819 ◽  
Author(s):  
D M Obzansky ◽  
K E Richardson
Keyword(s):  
Horseradish Peroxidase ◽  
Isotope Dilution ◽  
Reference Interval ◽  
Oxalate Oxidase ◽  
Exchange Chromatography ◽  
Urinary Oxalate ◽  
Cation Exchange Chromatography ◽  
Standard Curve ◽  
The Difference

Abstract We describe an automated (ABA-100) enzymic method for determination of urinary oxalate by use of oxalate oxidase (EC 1.2.3.4) isolated from beet stems. The H2O2 produced by the oxidation of oxalate by oxalate oxidase is measured by coupling with oxidation and conjugation of 3-methyl-3-benzothiazolinone hydrazone with N,N-dimethylaniline with catalysis by horseradish peroxidase. The resulting indamine dye is measured spectrophotometrically by the difference in absorption at 500 and 600 nm. Interfering substances are removed by oxidation with acidic ferric chloride and by cation-exchange chromatography. The assay is sensitive to 5 mg of urinary oxalate per liter, the standard curve is linear to 70 mg/L, and the procedure requires less than 3 h for completion. The within-run CV was less than 1.6%, the between-day CV less than 5.6%. The oxalate oxidase method results in a mean and reference interval for oxalate excretion that are comparable with those by isotope dilution, gas-chromatographic, colorimetric, and other enzymic procedures.

scholarly journals Peroxidase and peroxidase-oxidase activities of isolated human myeloperoxidases

1987 ◽  
Vol 242 (3) ◽  
pp. 673-680 ◽  
Author(s):  
B E Svensson ◽  
K Domeij ◽  
S Lindvall ◽  
G Rydell
Keyword(s):  
Enzyme Activity ◽  
Cation Exchange ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Relative Molecular Mass ◽  
Cation Exchange Chromatography ◽  
Healthy Donors ◽  
Activity Measurements ◽  
Molecular Masses ◽  
Sites Of Action

Isolated neutrophils from healthy donors were used for the isolation of four highly purified forms of myeloperoxidase as determined by spectral (A430/A280 ratio 0.80-0.87) and enzyme-activity measurements. Although the myeloperoxidases exhibited different elution profiles on cation-exchange chromatography, gel filtration indicated similar relative molecular masses. When these forms were assayed for peroxidase and peroxidase-oxidase activities with several substrates, they all exhibited virtually the same specific activities. These results suggest that possible functional differences between the enzymes may be related to differences in their sites of action rather than to differences in enzyme activity. Myeloperoxidase from a patient with chronic myeloid leukaemia also revealed a similar heterogeneity on cation-exchange chromatography. However, this myeloperoxidase contained in addition one form with a lower and one form with a higher relative molecular mass, as indicated by gel-filtration chromatography.

scholarly journals Antibacterial activity in bovine lactoferrin-derived peptides.

1997 ◽  
Vol 41 (1) ◽  
pp. 54-59 ◽  
Author(s):  
K S Hoek ◽  
J M Milne ◽  
P A Grieve ◽  
D A Dionysius ◽  
R Smith
Keyword(s):  
Antibacterial Activity ◽  
Disulfide Bond ◽  
Exchange Chromatography ◽  
Single Step ◽  
Bovine Lactoferrin ◽  
Reverse Phase ◽  
Methionine Residue ◽  
Before And After ◽  
High Sequence Homology ◽  
Edman Sequencing

Several peptides sharing high sequence homology with lactoferricin B (Lf-cin B) were generated from bovine lactoferrin (Lf) with recombinant chymosin. Two peptides were copurified, one identical to Lf-cin B and another differing from Lf-cin B by the inclusion of a C-terminal alanine (lactoferricin). Two other peptides were copurified from chymosin-hydrolyzed Lf, one differing from Lf-cin B by the inclusion of C-terminal alanyl-leucine and the other being a heterodimer linked by a disulfide bond. These peptides were isolated in a single step from chymosin-hydrolyzed Lf by membrane ion-exchange chromatography and were purified by reverse-phase high-pressure liquid chromatography (HPLC). They were characterized by N-terminal Edman sequencing, mass spectrometry, and antibacterial activity determination. Pure lactoferricin, prepared from pepsin-hydrolyzed Lf, was purified by standard chromatography techniques. This peptide was analyzed against a number of gram-positive and gram-negative bacteria before and after reduction of its disulfide bond or cleavage after its single methionine residue and was found to inhibit the growth of all the test bacteria at a concentration of 8 microM or less. Subfragments of lactoferricin were isolated from reduced and cleaved peptide by reverse-phase HPLC. Subfragment 1 (residues 1 to 10) was active against most of the test microorganisms at concentrations of 10 to 50 microM. Subfragment 2 (residues 11 to 26) was active against only a few microorganisms at concentrations up to 100 microM. These antibacterial studies indicate that the activity of lactoferricin is mainly, but not wholly, due to its N-terminal region.

Three assays for glycohemoglobin compared

1995 ◽  
Vol 41 (2) ◽  
pp. 191-195 ◽  
Author(s):  
U Turpeinen ◽  
U Karjalainen ◽  
U H Stenman
Keyword(s):  
Cation Exchange ◽  
Correlation Coefficient ◽  
Reference Method ◽  
Hplc Method ◽  
Exchange Chromatography ◽  
The Other ◽  
Affinity Binding ◽  
Cation Exchange Chromatography ◽  
The Difference

Abstract Using 123 specimens, we compared the concordance of three different methods for determining glycohemoglobin (GHb): the Diamat (Bio-Rad Laboratories), an automated analyzer measuring HbA1c by cation-exchange chromatography; an assay with the IMx analyzer (Abbott Laboratories), based on boronate affinity binding; and an HPLC method measuring HbA1c by cation-exchange chromatography on a PolyCAT A column (PolyLC Inc.). The Pearson's correlation coefficient between PolyCAT A and Diamat was 0.900 +/- 0.038 (mean +/- 2 SD) and between PolyCAT A and IMx, 0.857 +/- 0.042. However, up to twofold differences were seen in some samples. The proportion of GHb was consistently lower with the PolyCAT A method than with the other two assays, apparently because of better separation of HbA1c from nonglycated coeluting forms of Hb. The difference in glycation percentage between the PolyCAT A and Diamat methods is 2-3% over the whole concentration range. These results point to the limitations of Diamat as a reference method to be used to calibrate other methods for determining HbA1c. Further, a switch from one method to another is likely to cause considerable problems in the clinical follow-up of certain patients.

Purification and Characterization of an Alkaline Protease from Bacillus alcalophilus

2012 ◽  
Vol 192 ◽  
pp. 285-288 ◽  
Author(s):  
Shan Shan Liu ◽  
Li Li Wang ◽  
Lin Yuan
Keyword(s):  
Alkaline Protease ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Purification And Characterization ◽  
Highly Active ◽  
Wide Range ◽  
Commercial Applications ◽  
Ph Range

The purification and characterization of an alkaline protease produced by Bacillus alcalophilus were investigated. The enzyme was purified in two steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by cation-exchange chromatography. The purified protease had a molecular mass of approximately 28 kDa, was highly active over an alkaline pH range of 10.0 to 11.0, and remained stable over a pH range of 7.0 to 12.0. The optimum temperature for the enzyme activity was found to be 40~60°C, while the thermotolerance of the enzyme was poor. Therefore, these characteristics of the protease indicate its potential for a wide range of commercial applications.

scholarly journals Liberation of tryptic fragments from caseinomacropeptide of bovine κ-casein involved in platelet function. Kinetic study

1990 ◽  
Vol 271 (1) ◽  
pp. 247-252 ◽  
Author(s):  
J Léonil ◽  
D Mollé
Keyword(s):  
Platelet Function ◽  
Acid Precipitation ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Rate Of Hydrolysis ◽  
The Difference ◽  
Cationic Exchange ◽  
Hydrolysis Of ◽  
Similar Kinetic

Carbohydrate-free caseinomacropeptide (CMP) was purified from rennet-hydrolysed caseinate by trichloroacetic acid precipitation and DEAE-TSK Fractogel-650 ion-exchange chromatography. To study the liberation of 106-112, 106-116 and 113-116 fragments from carbohydrate-free CMP involved in platelet function, a quantitative study was made on the rate of hydrolysis of the three peptidic bonds that are susceptible to the action of trypsin. Data were obtained from reverse-phase (Ultrabase column) and cationic-exchange (Mono S column) h.p.l.c. On the basis of the disappearance of substrate, kcat. and Km were respectively 3.95 s-1 and 0.2 mM. The two 111-112 and 112-113 bonds were split according to similar kinetic parameters (kcat. = 1.97 s-1, Km = 0.2 mM) and much faster than the 116-117 bond. The difference in susceptibility of the bonds can probably be attributed to the nature of residues flanking the primary proteolytic sites rather than to their accessibility to the proteinase. On the basis of our results the 106-116 fragment cannot be formed.

A high performance liquid chromatographic assay for the mycotoxin phomopsin A in lupin stubble

1987 ◽  
Vol 27 (1) ◽  
pp. 73 ◽  
Author(s):  
GR Hancock ◽  
P Vogel ◽  
DS Petterson
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
High Performance Liquid Chromatographic ◽  
Exchange Chromatography ◽  
Reverse Phase ◽  
Extraction And Purification ◽  
Wide Range ◽  
Total Analysis ◽  
The Mean ◽  
Liquid Chromatographic

An assay using high performance liquid chromatography to measure phomopsin A, the principal mycotoxin responsible for lupinosis is described. Samples of lupin stubble are extracted with methanol: water and purified by partitioning between n-butanol and water, chromatography on Amberlite XAD-2 and by cation exchange chromatography. The analysis is performed on a reverse phase C18 column using a methanol:water gradient and UV detection. The limit of detection for this procedure is 0.5 mg phomopsin A per kg stubble. Improvements to the extraction and purification procedures were made and total analysis time was reduced to 2 days. The mean (� s.d.) recovery for the purification procedure was 64.3 � 4.5% over a wide range of concentrations.

Sign in / Sign up

Export Citation Format

Share Document