摘要
以普鲁兰酶生产真菌YBJ10-2基因组DNA为模板,PCR扩增出普鲁兰酶的结构基因,将此基因插入载体pPIC9K,构成重组质粒pPIC9K-Pullulanase,转化毕赤酵母SMD1168,并通过G418平板培养基筛选高拷贝转化子,普鲁兰酶表达并分泌到胞外。该酶的表达受甲醇的严格调控和诱导,摇瓶培养中,当甲醇达到最佳诱导浓度3%,酶的最高表达量可达225U/mL。该酶能够长时间保持稳定,并具有较高的酶活力。重组毕赤酵母遗传稳定性良好。
With fungi YBJ10-2 genomic DNA as template, PCR amplified pullulanase structure gene, and inserted into the vector pPIC9K to the recombinant pPIC9K-Pullulanase. Transforming to Pichia yeast SMDl168, screening for high copy transformant by the G418 plate culture medium, pullulanase secretion expression into the extraceUu- lar. The enzyme is expressed by the strict regulation and induction of methanol, when methanol achieves the best inducer concentration 3%, the enzyme supreme expression up to 225U/mL. This enzyme can keep stable for a long time, and has high enzyme activity. Recombinant Pichia yeast genetic is stability.
出处
《食品与发酵科技》
CAS
2012年第6期24-27,共4页
Food and Fermentation Science & Technology
