Isolation,purification and identification of protein with cholesterolreducing activity from the supernatant of Lactobacillus reuteri
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摘要: 罗伊氏乳杆菌DSM122460无细胞上清发酵液（cell-free supernatant,CFS）具有较高的胆固醇移除能力,经初步鉴定有效成分为蛋白质。通过超滤浓缩、硫酸铵分级沉淀、DEAE Sepharose F.F阴离子交换层析进一步对该活性成分进行分离纯化。纯化后,其比活力达到61.6 U/mg,纯化倍数为32.4。经SDS-PAGE检测,样品达到电泳级纯,分子量约为60 ku。通过质谱鉴定初步认定该活性蛋白为一假设蛋白,其功能尚待进一步研究。Abstract: The cell- free supernatant( CFS) of Lactobacillus reuteri DSM122460 exhibited the cholesterol removal ability and the effective components were predicted as proteins. The active component was purified by ultrafiltration,ammonium sulfate precipitation,and DEAE- Sepharose F.F anion exchange chromatography. The specific activity of purified component was 61.6 U / mg with a purification factor of 32.4. The purified protein was electrophoresis pure determined by SDS- PAGE,and the estimated molecular weight was about 60 ku. The active protein was identified as a hypothetical protein by mass spectrometry and its function remains to be further studied.
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