荒漠昆虫小胸鳖甲抗冻蛋白MpAFP698对酿酒酵母的低温保护作用  

Cryoprotective effect of antifreeze protein MpAFP698 from the desertinsect Microdera punctipennis on yeast Saccharomyces cerevisiae

作　　者：张凤娟[1] 孙琳洁[1] 李素丽[1] 马纪[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：《食品与发酵工业》2017年第5期37-42,共6页Food and Fermentation Industries

基　　金：国家自然科学基金(31360527)

摘　　要：昆虫抗冻蛋白作为天然无毒副作用的低温和冷冻保护剂,在发酵菌种的保藏方面具有很好应用前景。为了验证荒漠昆虫小胸鳖甲(Microdera punctipennis)抗冻蛋白Mp AFP698对酵母菌的抗冻和抗低温保护效果,对Mp AFP698抗冻蛋白进行原核表达和纯化,获得了具有活性的融合抗冻蛋白MBP-Mp AFP698。将质量浓度为1、10、30μg/m L的MBP-Mp AFP698添加到酿酒酵母(Saccharomyces cerevisiae)中,在-7和-20℃冷冻处理酵母菌液不同时间;以10μg/m L的抗冻蛋白在4、0、-50℃处理酵母菌液,在-20℃反复冻融处理,观察菌落形态并统计菌种存活率。结果表明,添加抗冻蛋白组的酵母存活率显著优于未加抗冻蛋白的对照组,1μg/m L就有保护效果,10μg/m L保护效果最好,说明昆虫抗冻蛋白能以极低的浓度发挥低温保护作用。Antifreeze proteins as non-toxic cryoprotective agents have extensive prospect in fermentation strain preservation. To test the cryoprotective effect of antifreeze protein MpAFP698 from the desert beetle Microdera puncti- pennis on S. cerevisiae, antifreeze protein MpAFP698 was prokaryotic expressed and purified. Fusion protein MBP- MpAFP698 with high activity was obtained. 1 , 10, 30 μg/mL of MBP-MpAFP698 were added to S. cerevisiae and then treated at -7 and -20℃ for different time lengths; in other experiments yeast cells were treated at 4, 0, -50 ℃ for different time lengths with 10 μg/mL MpAFP698 as well as repeated freeze-thaw at -20℃. The mor- phology and survival rate of the yeast cells after these treatments were observed or calculated. The results showed that the survival rate of S. cerevisiae with the protection of MBP-MpAFP698 were significantly better than the control group without MBP-MpAFP698. Even the concentration of 1 μg/mL showed protective effect, and 10 μg/mL showed the best protective effect, indicating that insect antifreeze protein can function as eyroprotective agent at very low concen- trations. This result lay a foundation for the development of efficient protective agent at low temperatures.

关 键 词：荒漠昆虫抗冻蛋白 酿酒酵母 抗冻保护 细胞存活 

分 类 号：TQ926[轻工技术与工程—发酵工程]