Purification and characterization of a approximately 34 kDa antioxidant protein (beta-turmerin) from turmeric (Curcuma longa) waste grits.
Beta-turmerin from turmeric (Curcuma longa) waste grits obtained after extraction of curcumin was purified by successive gel permeation chromatography. Homogeneity of beta-turmerin was confirmed by its movement as single band both in SDS-PAGE and as well as in native (basic) PAGE. The apparent molecular mass is approximately 34 kDa by SDS-PAGE. It is more hydrophobic protein and showed sharp single peak in RP-HPLC with retention time of 62.17 min. It is a glycoprotein as it shows the presence of amino sugars up to 0.021 gm%. In three different model systems i.e., linolenic acid micelles, erythrocyte membrane systems and liposomes, beta-turmerin at 0.125 microM offered 70%, 64%, and 60% inhibition of lipid peroxidation, which is 3200 times more efficient than the standard antioxidants BHA (400 microM) and alpha-tocopherol (400 microM). beta-turmerin inhibited diene-triene and tetraene conjugation up to 54%, 72% and 47%, respectively. beta-turmerin also effectively scavenges hydroxyl radicals when compared to BHA and alpha-tocopherol. beta-turmerin (2.5 microM) further inhibited the activation of PMNL mediated by fMLP up to the extent of 75%, where as standards BHA (400 microM) and mannitol (10 microM) inhibited the same to 65% and 55%, respectively. At 0.125 microM dose beta-turmerin prevented t-BOOH induced cell death at all time intervals. In addition to the above properties, it is non-toxic to lymphocytes as it did not affect the viability of cells. The mechanism of antioxidant action of beta-turmerin could probably be by counteracting/quenching of reactive oxygen species (ROS). We report the purification and characterization of beta-turmerin ( approximately 34 kDa), a potent antioxidant protein from turmeric waste grits.
Smitha
S
S
Adichunchanagiri Biotechnology and Cancer Research Institute, Balagangadharanatha Nagara, Mandya District, Karnataka 571 448, India.
Dhananjaya
B L
BL
Dinesha
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R
Srinivas
Leela
L
D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D002084:Butylated Hydroxytoluene / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D030024:Curcuma / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D000975:Antioxidants; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D010936:Plant Extracts; D010940:Plant Proteins; D017382:Reactive Oxygen Species; C072268:turmerin; D002084:Butylated Hydroxytoluene; C012596:turmeric extract