新疆家蚕抗菌肽在酿酒酵母中表达的实时监测及活性分析  

Real-time monitoring the Cecropin XJ expression in Saccharomyces cerevisiae and evaluation of antibacterial activity

作　　者：刘东亮[1] 刘军[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《食品工业科技》2016年第17期129-133,共5页Science and Technology of Food Industry

基　　金：新疆维吾尔自治区高新技术研究和发展项目(201110101);新疆生物资源基因工程重点实验室开放基金(XJDX0201-2014-02)

摘　　要：本研究对新疆家蚕抗菌肽在酿酒酵母INVSc1菌株中分泌表达过程进行了实时监测及活性测定。结果表明,选择培养基中酵母表现为典型的S型生长曲线;诱导培养基中酵母表现为波浪式缓慢上升的曲线,上升的时间和速度与酵母的初始接菌密度有关。诱导过程补加少量20%半乳糖诱导剂可以明显抑制酵母的生长,同时,酵母细胞平板计数结果也表明补加半乳糖使酿酒酵母出芽增殖减缓,菌落计数减少。研究发现诱导过程补加半乳糖不仅可以使发酵液中总蛋白的含量提高约50%,还可以使酵母分泌蛋白的时间提前。抗菌实验表明,抗菌肽对大肠杆菌抑菌效果较好,发酵液上清对大肠杆菌的最大抑菌圈直径为52 mm;对金黄色葡萄球菌的抑菌活性较弱,最大抑菌圈直径为20 mm。In the present study, the expression process of Cecropin XJ in INVScl strain of Saccharomyces cerevisiaewas real-timely monitored. Firstly, the growth states of yeast in SD selective medium （Ura-）and induction medium were measured, respectively.The result showed that a sigmoid growth curve and a wave-like rising curve were respectively observed in selective medium and induction medium.During the induction process, the rising rate of growth curve was related to the cell density at the beginning of induction.Furthermore,the growth of yeast was obviously suppressed after refilling some 20% galactose in induction culture.Similarly,the plate colony -counting assay indicated that galactose supplement lead to the decrease of yeast proliferation and colony numbers during the induction phase.Additionally, galactose supplement not only increased the protein content by 50% in the culture, but accelerated the protein secretion from yeast. The antibacterial assay revealed that the expression product exerted a significantly bactericidal ability against E. coil with the biggest inhibitory zone diameter of 52 mm,and a weak ability against S.aureus with a inhibitory zone diameter of 20 mm.

关 键 词：新疆家蚕抗菌肽 酿酒酵母 蛋白表达 抑菌活性 

分 类 号：TS201.1[轻工技术与工程—食品科学]