摘要
利用碱溶酸沉法从魔芋飞粉中提取魔芋蛋白,以魔芋蛋白为原料,利用碱性蛋白酶酶解制备魔芋ACE抑制肽粗品;其粗品通过超滤法除去大分子杂质,再经过Sephadex G-15柱层析分离,最后经过RP-HPLC纯化后得到纯度较高的魔芋ACE抑制肽,并且对超滤膜进行选择、Sephadex G-15柱层析分离进行条件优化;以马尿酸含量为指标,紫外分光光度法测其活性。实验结果表明:选择规格为1 kDa超滤膜;Sephadex G-15最佳分离条件为浓度为120 mg/mL、流速为0.8 mL/min、上样量为1.0 mL;经过RP-HPLC纯化后得到魔芋ACE抑制肽抑制率可达到92.85%。空白液中Hip含量为22.50 mg,反应液中Hip含量为9.22 mg,说明魔芋ACE抑制肽抑制活性较好。
Protein was extracted from konjac powder by alkali extraction and acid precipitation method; konjac protein is material, konjac ACE inhibitory peptides crude were prepared by alkaline hydrolysis protease on konjac pro- tein; its crude macromolecular impurities were removed by uhrafiltration, Sephadex G-15 column chromatography, and RP-HPLC purification. Highly purified konjac ACE inhibitory peptide was obtained. The ultrafiltration, Sephadex G-15 column chromatographic separation conditions were optimized; hippuric acid was used as the index and Spectro- photometric was used to determine the activity. The results showed that an ultrafihration membrane has a molecular weight of 1 kDa, The optimal separation condition for Sephadex G-15 was 120 mg/mL, flow rate was 0.8 mL/min, sample volume was 1.0 mL. After RP-HPLC purified, konjac ACE inhibitory peptide inhibition rate was reached to 92.85%. Blank solution Hip content was 22.50 mg, the reaction mixture Hip content was 9.22 mg, which indicates that konjac ACE inhibitory peptide inhibitory activity was better.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2017年第6期163-168,共6页
Food and Fermentation Industries
基金
陕西省科技攻关项目(2016NY-167)
西安市科技计划项目(NC1405(3))
陕西省农业科技创新与攻关项目(2015NY010
2016NY-142)
陕西省协同创新计划项目(2016XT-21)
关键词
魔芋蛋白
ACE抑制肽
分离纯化
活性
konjac protein
ACE inhibitory peptides
separation and purification
activity
