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Antioxidant, antibacterial, and cytoprotective activity of agathi leaf protein.

Zarena AS, Gopal S, Vineeth R - J Anal Methods Chem (2014)

Bottom Line: The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical.The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min.In addition, ALP showed strong antibacterial activity against Pseudomonas aeruginosa (20 ± 3.64 mm) and Staphylococcus aureus (19 ± 1.53 mm) at 200  μ g/mL.

View Article: PubMed Central - PubMed

Affiliation: Department of Studies in Microbiology, University of Mysore, Mysore 570006, India.

ABSTRACT
In the present study a protein termed agathi leaf protein (ALP) from Sesbania grandiflora Linn. (agathi) leaves was isolated after successive precipitation with 65% ammonium sulphate followed by purification on Sephadex G 75. The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical. SDS-PAGE analysis of P I indicated that the molecular weight of the protein is ≈ 29 kDa. The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min. ALP was able to reduce oxidative damage by scavenging lipid peroxidation against erythrocyte ghost (85.50 ± 6.25%), linolenic acid (87.67 ± 3.14%) at 4.33  μ M, ABTS anion (88 ± 3.22%), and DNA damage (83 ± 4.20%) at 3.44  μ M in a dose-dependent manner. The purified protein offered significant protection to lymphocyte (72% at 30 min) induced damage by t-BOOH. In addition, ALP showed strong antibacterial activity against Pseudomonas aeruginosa (20 ± 3.64 mm) and Staphylococcus aureus (19 ± 1.53 mm) at 200  μ g/mL. The safety assessment showed that ALP does not induce cytotoxicity towards human lymphocyte at the tested concentration of 0.8 mg/mL.

No MeSH data available.


Related in: MedlinePlus

Inhibition of hydroxyl radical-mediated DNA degradation by ALP. Data are expressed as the mean ± standard deviation (n = 3). Means with different letters (a–c) are significantly different (P < 0.05).
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fig3: Inhibition of hydroxyl radical-mediated DNA degradation by ALP. Data are expressed as the mean ± standard deviation (n = 3). Means with different letters (a–c) are significantly different (P < 0.05).

Mentions: The effect of ALP on hydroxyl radicals generated by Fe3+/H2O2 ions was measured by determining the degree of DNA degradation by test tube assay (Figure 3). While a marginal inhibition was evident at the lower concentration, nearly 83 ± 4.20% inhibition was observed at higher concentration at 3.44 μM. The scavenging effect increased with increasing ALP concentration up to a certain extent (3.44 μM) and then leveled off with further increase. Further ABTS●+ method showed an activity of 88 ± 3.22% at 3.44 μM when compared to synthetic antioxidant BHA which showed an activity of 92 ± 4.03% at 400 μM (Figure 4) and leveled off thereafter. It was observed that the inhibition value of ALP increased with increase in concentration.


Antioxidant, antibacterial, and cytoprotective activity of agathi leaf protein.

Zarena AS, Gopal S, Vineeth R - J Anal Methods Chem (2014)

Inhibition of hydroxyl radical-mediated DNA degradation by ALP. Data are expressed as the mean ± standard deviation (n = 3). Means with different letters (a–c) are significantly different (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3927847&req=5

fig3: Inhibition of hydroxyl radical-mediated DNA degradation by ALP. Data are expressed as the mean ± standard deviation (n = 3). Means with different letters (a–c) are significantly different (P < 0.05).
Mentions: The effect of ALP on hydroxyl radicals generated by Fe3+/H2O2 ions was measured by determining the degree of DNA degradation by test tube assay (Figure 3). While a marginal inhibition was evident at the lower concentration, nearly 83 ± 4.20% inhibition was observed at higher concentration at 3.44 μM. The scavenging effect increased with increasing ALP concentration up to a certain extent (3.44 μM) and then leveled off with further increase. Further ABTS●+ method showed an activity of 88 ± 3.22% at 3.44 μM when compared to synthetic antioxidant BHA which showed an activity of 92 ± 4.03% at 400 μM (Figure 4) and leveled off thereafter. It was observed that the inhibition value of ALP increased with increase in concentration.

Bottom Line: The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical.The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min.In addition, ALP showed strong antibacterial activity against Pseudomonas aeruginosa (20 ± 3.64 mm) and Staphylococcus aureus (19 ± 1.53 mm) at 200  μ g/mL.

View Article: PubMed Central - PubMed

Affiliation: Department of Studies in Microbiology, University of Mysore, Mysore 570006, India.

ABSTRACT
In the present study a protein termed agathi leaf protein (ALP) from Sesbania grandiflora Linn. (agathi) leaves was isolated after successive precipitation with 65% ammonium sulphate followed by purification on Sephadex G 75. The column chromatography of the crude protein resulted in four peaks of which Peak I (P I) showed maximum inhibition activity against hydroxyl radical. SDS-PAGE analysis of P I indicated that the molecular weight of the protein is ≈ 29 kDa. The purity of the protein was 98.4% as determined by RP-HPLC and showed a single peak with a retention time of 19.9 min. ALP was able to reduce oxidative damage by scavenging lipid peroxidation against erythrocyte ghost (85.50 ± 6.25%), linolenic acid (87.67 ± 3.14%) at 4.33  μ M, ABTS anion (88 ± 3.22%), and DNA damage (83 ± 4.20%) at 3.44  μ M in a dose-dependent manner. The purified protein offered significant protection to lymphocyte (72% at 30 min) induced damage by t-BOOH. In addition, ALP showed strong antibacterial activity against Pseudomonas aeruginosa (20 ± 3.64 mm) and Staphylococcus aureus (19 ± 1.53 mm) at 200  μ g/mL. The safety assessment showed that ALP does not induce cytotoxicity towards human lymphocyte at the tested concentration of 0.8 mg/mL.

No MeSH data available.


Related in: MedlinePlus