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An effective and simple procedure to isolate abundant quantities of biologically active chemopreventive Lunasin Protease Inhibitor Concentrate (LPIC) from soybean

Hari B. Krishnan, Thomas T.Y. Wang
Food chemistry 2015 v.177 pp. 120-126
immunoblotting, anti-inflammatory activity, soy flour, albumins, humans, soy protein, calcium, proteinases, chemical precipitation, chemoprevention, cytokines, soybeans, purification methods, mass spectrometry, protein content, clinical trials, ethanol, Kunitz-type proteinase inhibitor, anticarcinogenic activity
Lunasin is a 5-kDa soybean bioactive peptide with demonstrated anti-cancer and anti-inflammatory properties. Recently, purification methods have been developed to obtain gram quantities of lunasin. However, these methods are cumbersome, time consuming and cost-prohibitive. To overcome these constrains we have developed a novel method which involves extraction of soybean flour with 30% ethanol followed by preferential precipitation of lunasin and protease inhibitors by calcium. The calcium precipitated protein fraction, which we termed as Lunasin Protease Inhibitor Concentrate (LPIC), contains three abundant proteins with molecular weights of 21, 14 and 5kDa. This simple procedure yields 3.2g of LPIC from 100g of soybean flour and the entire isolation procedure can be completed in less than 2h. Treatment of THP-1 human monocyte cell lines with LPIC resulted in suppression of lipopolysaccharide-stimulated cytokine expression, demonstrating that the LPIC isolated by our simple procedure is biologically active.