【作者】 扈进冬； 郭勇； 吴远征； 张广志； 杨合同；

【Author】 HU Jin-dong,GUO Yong,WU Yuan-zheng,ZHANG Guang-zhi,YANG He-tang (Biotechnology Research Center of Shandong Academy of Sciences,Key laboratory of Applied Microbiology of Shandong Province,Jinan 250014,China; Insititute of Biology,Shandong Academy of Sciences,Jinan 250014,China; School of Life Science,Shandong University of Technology,Zibo 255049,China )

【机构】 山东省科学院中日友好生物技术研究中心山东省应用微生物重点实验室；

【摘要】 Cecropins B是一种昆虫抗菌肽,具有分子量小,热稳定性、水溶性好、抗菌谱广等优点,更为重要的是抗菌肽对真核细胞几乎没有作用,仅仅作用于原核细胞和发生病变的真核细胞,是目前已知天然抗菌肽中活力最强的一种。根据毕赤氏巴斯德酵母(Pichia pastoris)偏好密码子,改造并化学合成天蚕素基因B,克隆到 pPIC9K载体中,构建分泌型重组酵母表达载体CB-pPIC9K,转化Pichia pastoris受体菌KM71,在醇氧化酶(AOX) 启动子调控下,Cecropin B获得表达,分子量约3．8KD,摇瓶发酵产率可达到200μg/mL,并对其抗菌活性进行了初步测定。结果表明我们表达的重组Cecropins B对G-菌有较好的抑菌活性,且具有较好的热稳定性。这些特点使得重组抗菌肽Cecropins B在食品防腐、疾病防治和动物饲料添加剂等方面显露出很好的应用前景。更多还原

【Abstract】 Cecropins B is a kind of insect antibacterial peptide, which have the lower molecular weight , thermostability, water-solubility good and have bactericidal activity against a broad range of bacteria. Especially, nearly no have the function to the eukaryotic cell . At present, cecropins B is the activities strongest one kind in which is known nature’s antibacterial peptide. According to the partiality codon of Pichia pastoris, antibacterial peptides cecropin B gene was reconstructed, synthesized and cloned into pPIC9K to construct the recombinant expression vector CB-pPIC9K. The CB-pPIC9K was transformed into P. pastoris KM71. Under the control of the promoter AOX, cecropinB protein was expressed with the high level of 200μg/mL in the flask-shaking culture fermentation. In addition, the cecropinB protein with antibacterial activity was analyzed. The results indicated which the cecropins B has the good antibactrial activities to grem-negative bacterial and the thermostability. These characteristics of cecropins B display very good application prospect in the field of antisepsis of food anticorrosion, disease preventing and additives of animal feedstuff and so on.更多还原