乳酸菌盐溶性蛋白培养物中血管紧张素转化酶抑制活性肽的测定及分离

Separation and Determination of Angiotensin Converting Enzyme Inhibitory Peptide from Salt-Soluble Protein Solution Fermented with Lactic Acid Bacteria

将16株乳酸菌用MRS液体培养基3代继代培养后,离心,并用灭菌的生理盐水制成乳酸菌悬浮液,接种于远东多线鱼肉盐溶性蛋白溶液(salt-soluble protein,SSP),对其代谢产物进行血管紧张素转化酶(angiotensin converting enzyme,ACE)抑制活性测定,筛选出ACE抑制活性较高的Pediococcus acidilactici ID7菌株(ACE抑制率为47.6%)和Lactobacillus plantarum 6214菌株(ACE抑制率为40.6%)。利用高效液相色谱(high performance liquid chromatography,HPLC)对Pediococcus acidilactici ID7的盐溶性蛋白培养代谢物进行色谱分析,获得了两个峰,其相应成分对ACE抑制的IC50分别为1.21μg/m L和1.07μg/m L,利用凝胶过滤HPLC法对出现较高的ACE抑制活性峰的成分进行色谱纯化,获得了ACE抑制率为26.67%,分子质量为586.7 D以下的ACE抑制多肽。

Sixteen lactic acid bacteria（LAB） strains were cultured for 3 generations in MRS liquid medium, centrifuged, prepared into LAB suspension with sterilized physiological saline, and then inoculated in salt-soluble protein（SSP） solution from arabesque greenling. Through determining angiotensin converting enzyme（ACE） inhibitory peptide from metabolites, Pediococcus acidilactici ID7 and Lactobacillus plantarum 6214 displayed high ACE inhibitory activity, with a percentage inhibition of 47.6% and 40.6%, respectively. Higher ACE inhibitory activity in SSP solution fermented with Pediococcus acidilactici ID7 was determined by high performance liquid chromatograph（HPLC）. Meanwhile, two peaks after HPLC purification were achieved, which showed IC50 of 1.21 μg/m L and 1.07 μg/m L, respectively. The peak with higher ACE inhibitory activity was purified by gel filtration. The obtained ACE inhibitory peptide showed percentage inhibition of 26.67% and molecular weight less than 586.7 D.