乳源降血压七肽基因在毕赤酵母中的表达研究

利用基因工程技术在毕赤酵母中高效表达乳源抗高血压七肽,为大规模生产抗高血压肽奠定基础。根据毕赤酵母密码子偏爱性,人工合成了具有较高活性的抗高血压七肽串联基因序列。将串联基因克隆至表达载体pPIC9上,并转化毕赤酵母GS115,经PCR检测获得2株重组菌株。重组菌经甲醇诱导,表达的分泌蛋白经胰蛋白酶酶解,测定酶解产物抑制ACE酶活性。结果表明,表达蛋白的胰蛋白酶酶解产物抑制ACE酶活性达到65.12%。IC50值为0.0203 mg.mL-1。研究成功合成了串联的13拷贝抗血压七肽基因,并实现了其在毕赤酵母中的高效表达,表达蛋白的胰蛋白酶酶解产物具有抗高血压肽活性。

Dairy-derived anti-hypertension seven peptide tandem gene synthesis and expression in Pichia pastoris

This research used genetic engineering technology in Pichia efficiently express in milk source antihypertensive peptides for mass production,and seven antihypertensive peptides lay the foundation.According to the Pichia anticodon preference,synthetic with highly antihypertensive peptides seven series gene sequences,and further series for 13 copy antihypertensive peptides,cloning and expression vector pPIC9 and GS115 Pichia transformation.Recombinant strains,cells induced by methanol,expressed the trypsin digestion,and protein determination of enzymatic hydrolysis product inhibiting enzymes ACE.Expression of proteins inhibit trypsin digestion products to 65.12% ACE enzyme activity and IC50 value concentration for 0.0203 mg·mL-1.The detection of recombinant strains had two strains.The success of the 13 series synthetic peptide gene copies resistance,and blood pressure seven in Pichia realized the high protein expression,expression of trypsin digestion products had antihypertensive peptides activity.