蛇毒素蛋白毕赤酵母表达进展  

Advances on Expression of Snake Venom Protein in Pichia pastoris

作　　者：杜国俊[1] 刘晓飞[1] 王红军[2] 杨章民[1] 

机构地区：[1]陕西师范大学生命科学学院,西安710062 [2]哈佛大学医学院Dana-Farber癌症研究所,波士顿02115

出　　处：《中国生物工程杂志》2010年第10期86-93,共8页China Biotechnology

基　　金：国家自然科学基金(30870303);陕西省自然科学基金(SJ08C102);中央高校基本科研业务费专项资金(GK200902029)资助项目

摘　　要：蛇毒是许多具有独特生物活性的蛋白质与酶的混合物,在基础科学研究和临床上有重大应用价值,但是通过从蛇毒中分离获取活性组分具有局限性。巴斯德毕赤酵母表达系统是最为常用的真核表达系统之一,其真核加工、折叠、翻译后修饰等能力使得所表达的重组蛋白具有与天然蛋白近似的生物活性,因而该系统在富含二硫键或糖基化的蛇毒素蛋白表达中被广为采用。迄今为止,已经有12个属的25种蛇毒素蛋白(包括蛇毒丝氨酸蛋白酶、金属蛋白酶/去整合素、L-氨基酸氧化酶、C-型凝集素和神经毒素、血管收缩因子、神经生长因子等家族)在毕赤酵母中获得成功表达,蛇毒富半胱氨酸蛋白、缓激肽增强肽(BPP)等至今尚未见酵母表达的报道。毕赤酵母表达蛇毒素蛋白失败的原因可能在于,有关密码子偏爱性、目的基因转录出的RNA二级结构特征、糖基化程度不均一及糖型差异、所表达毒素对酵母细胞的毒性等方面,并对解决的方法进行了讨论。Snake venom is a mixture of many proteins and enzymes with distinct biological activities,which may have significant values both in basic researches and clinical applications.However,limitation exists in the purification of sufficient active fractions for practical use.Pichia pastoris is the most commonly used eukaryotic expression system nowadays.The ability of eukaryotic processing,folding and post-translational modification endow the recombinant proteins expressed with similar biological activities of the corresponding natural proteins,is thus widely adopted to express snake venom proteins with glycosylation or rich in disulfide bonds.To date,about twenty-five snake venom proteins of 12 genus have been expressed successfully in Pichia pastoris,（including different snake venom family members such as serine proteinases,metalloproteinases/disintegrins,L-amino acid oxidases,C-type lectin like proteins,neurotoxins,vasoconstrictors,nerve growth factor,etc.）,so far,snake venom cysteine-rich protein,Bradykinin-potentiating peptides etc.expressed in Pichia pastoris have not been reported.The reasons for the failure of expression of snake venom protein in Pichia pastoris maybe as follows：the codon bias,RNA secondary structure of the gene transcripted,heterogeneity of glycosylation and the difference between glycosylation types,the toxicity of expressed toxic protein on yeast cells etc.,The measures to these problems are discussed also.

关 键 词：蛇毒 蛋白家族 毕赤酵母表达 

分 类 号：Q786[生物学—分子生物学]