Purification of Bovine Milk Lactoperoxidase and Investigation of Antibacterial Properties at Different Thiocyanate Mediated

Uğuz, Metin Tansu; Özdemir, Haşan

doi:10.1007/s10438-005-0059-8

Cited by 20 publications

(18 citation statements)

References 20 publications

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“…7), using TMB as a substrate as described in Materials and Methods, while the K m for MPO and LPO are 0.5 mM and 0.2 mM, respectively [34, 35]. The data indicate that the K m of VPO1 is about 3-fold and 7.5-fold higher than MPO and LPO, respectively.…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of VPO1, a new animal heme-containing peroxidase

Cheng¹,

Salerno²,

Cao³

et al. 2008

Free Radical Biology and Medicine

118

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Animal heme-containing peroxidases play roles in innate immunity, hormone biosynthesis and the pathogenesis of inflammatory diseases. Using the peroxidase-like domain of Duox1 as a query, we carried out homology searching of the NCBI database. Two novel heme-containing peroxidases were identified in humans and mice. One, termed VPO1 (vascular peroxidase 1), shows highest tissue expression in heart and vascular wall. A second, VPO2, present in humans but not in mice, is 63% identical to VPO1, and is highly expressed in heart. The peroxidase-homology region of VPO1 shows 42% identity to myeloperoxidase (MPO) and 57% identity to insect peroxidase, peroxidasin. A molecular model of VPO1 peroxidase region shows a structure that is highly similar to known peroxidases, including a conserved heme-binding cavity, critical catalytic residues, and a calcium-binding site. Absorbance spectra of VPO1 are similar to lactoperoxidase and covalent attachment of the heme to VPO1 protein was demonstrated by chemiluminescent heme staining. VPO1 purified from heart or expressed in HEK cells is catalytically active and shows a Km for H2O2 of 1.5 mM. When co-expressed in cells, VPO1 can utilize H2O2 produced by Nox enzymes. VPO1 is likely to carry out peroxidative reactions in the vascular system previously attributed exclusively to MPO.

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Section: Resultsmentioning

confidence: 99%

Identification and characterization of VPO1, a new animal heme-containing peroxidase

Cheng¹,

Salerno²,

Cao³

et al. 2008

Free Radical Biology and Medicine

118

View full text Add to dashboard Cite

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“…They are capable of reducing bacterial growth by damaging cell membranes and inhibiting activities of several cytoplasmic enzymes. The biocidal activity of the LPO results from the products of the chemical reactions that it catalyzes . Hypothiocyanite, which is the main product of the reaction, interacts with thiol groups of various proteins, which are critical for the survival of pathogens.…”

Section: Discussionmentioning

confidence: 99%

“…[49] These compounds are oxidized in end products that are safe for human The biocidal activity of the LPO results from the products of the chemical reactions that it catalyzes. [50] Hypothiocyanite, which is the main product of the reaction, interacts with thiol groups of various proteins, which are critical for the survival of pathogens. The impact of LPO on bacteria results from the oxidation of sulfhydryl.…”

Section: Discussionmentioning

confidence: 99%

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The inhibition effects of some natural products on lactoperoxidase purified from bovine milk

Köksal

Kalın

Gerni

et al. 2017

J Biochem Mol Toxicol

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In this study, inhibition profiles of some natural products, which are digoxin, L-Dopa, dopamine, isoliquiritigenin, and 1,1,2,2-tetrakis(p-hydroxyphenyl)ethane (Tetrakis), were investigated against bovine lactoperoxidase (LPO) enzyme. Digoxin, L-Dopa, and dopamine are active ingredients of some drugs, which have important functions in our body, especially in cases of heart failure. Isoliquiritigenin and tetrakis are types of natural phenolic compounds, which play an important role in cancer prevention and treatment. LPO enzyme was purified from bovine milk using sepharose-4B-l-tyrosine sulfonamide affinity column chromatography. LPO is responsible for the nonimmune biological defense system and has antibacterial activity so selection of these active substances is important. The inhibition studies are performed with the ABTS substrate. Bovine LPO enzyme was effectively inhibited by phenolic molecules. K values of these natural products were found as 0.20 ± 0.09, 0.22 ± 0.17, 0.49 ± 0.11, 0.49 ± 0.27, and 1.20 ± 0.25 μM, respectively. Tetrakis and digoxin exhibited noncompetitive inhibition, and other molecules showed competitive inhibition.

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“…Thereafter, the enzyme solution was dialyzed overnight against phosphate buffer (5 mM, pH: 6.8). [21,22] LPO was obtained from the CM-Sephadex C-50 column, which was applied to a column of Sephadex G-100 (2.5 × 100 cm). The column-bound enzyme was eluted with 100 mM phosphate buffer (pH: 6.8), and salted out with ammonium sulphate precipitation (III Precipitation, 90% saturation).…”

Section: Purification Of Lactoperoxidasementioning

confidence: 99%

The Inhibitory Effects of L-Adrenaline on Lactoperoxidase Enzyme Purified from Bovine Milk

Şişecioğlu

Gülçın

Çankaya

et al. 2012

International Journal of Food Properties

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L-Adrenaline belongs to a group of compounds known as catecholamines, which play an important role in the regulation of the physiological process in living organisms. In the present study, the inhibitory effect of L-adrenaline on lactoperoxidase was examined. Lactoperoxidase (E.C.1.11.1.7) was purified from bovine milk with three consecutive steps: Amberlite CG-50 resin, CM-Sephadex C-50 ion-exchange, and Sephadex G-100 gel filtration chromatography. Lactoperoxidase was purified with a yield of 42.18%, a specific activity of 30.33 EU/mg proteins, and 20.77 purification fold. Enzyme purity was determined with SDS-PAGE, where a single band was observed. The R z (A 412 /A 280 ) value for lactoperoxidase was 0.9. The effect of L-adrenaline on lactoperoxidase was determined using ABTS as a chromogenic substrate. The half maximal inhibitory concentration (IC 50 ) value and an inhibition constant (K i ) values for L-adrenaline were 34.5 and 2.26 µM, respectively. L-Adrenaline was found to be a non-competitive inhibitor.

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Purification of Bovine Milk Lactoperoxidase and Investigation of Antibacterial Properties at Different Thiocyanate Mediated

Cited by 20 publications

References 20 publications

Identification and characterization of VPO1, a new animal heme-containing peroxidase

Identification and characterization of VPO1, a new animal heme-containing peroxidase

The inhibition effects of some natural products on lactoperoxidase purified from bovine milk

The Inhibitory Effects of L-Adrenaline on Lactoperoxidase Enzyme Purified from Bovine Milk

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