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| 超耐热酸性α-淀粉酶基因在多型汉逊酵母中表达及与在毕赤酵母中表达的比较研究 | |
| 引用本文： | 郝帅,郭建强,李运敏,岳丽丽,矫庆华.超耐热酸性α-淀粉酶基因在多型汉逊酵母中表达及与在毕赤酵母中表达的比较研究[J].菌物学报,2006,25(4):573-578. |
| 作者姓名： | 郝帅 郭建强 李运敏 岳丽丽 矫庆华 |
| 作者单位： | 1. 中国科学院微生物研究所,北京,100080;中国科学院研究生院,北京,100049 2. 三元基因有限公司,北京,100083 3. VA Medical Centre 111C5,Department of Medicine University of California at San Francico 4150 Clement Street,San Francisco,CA 94121,USA 4. 中国科学院微生物研究所,北京,100080 |
| 摘 要： | 利用毕赤酵母的质粒载体pPIC9K将极端耐热古菌Pyrococcusfuriosus的超耐热酸性α-淀粉酶(Amy)基因转化到多型汉逊酵母HP-6中,获得重组汉逊酵母。经过甲醇进行诱导,表达产物的酶活性检测和SDS-PAGE电泳,证明α-淀粉酶(Amy)在多型汉逊酵母中利用AOX1启动子和α-因子信号肽有效表达并分泌到胞外。该酶的最适反应温度为90～100℃,最适作用pH为4.0～5.0,较之重组毕赤酵母的最适作用pH还低0.5。此外与毕赤酵母的重组蛋白相比,重组汉逊酵母α-淀粉酶不仅菌株筛选简便、周期短,而且具有更容易筛选到高拷贝转化子以及适用于大规模工业发酵等优点。 |
| 关 键 词： | Pyrococcusfuriosus AOX1启动子 MOX启动子 α-因子信号肽 |
| 文章编号： | 1672-6472（2006）04-0573-0578 |
| 收稿时间： | 2006-05-15 |
| 修稿时间： | 2006-05-23 |
Comparison between the expression of extremely thermostable and acid-stable amylase gene in Hansenula polymorpha and that in Pichia pastoris |
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| Authors: | HAO Shuai GUO Jian-Qiang LI Yun-Min YUE Li-Li JIAO Qing-Hua |
| Institution: | 1 Institite of Microbiology, Chinese Academy of Sciences, Beijing 100080, China; 2Graduate School, Chinese Academy of Sciences, Beijing 100049, China; 3Sanyuan Gene Limited Company, Beijing 100083, China; 4VA Medical Centre 111C5, Department of Medicine University of California at San Franeico 4150 Clement Street, San Francisco, CA 94121, USA |
| Abstract: | A recombinant vector pPIC9K-Amy which carries the extremely thermostable and acid-stable α-amylase from Pyrococcus furiosus was transformed into Hansenula polymorpha HP-6 strain. Regulated by promotor of AOX1 gene and the α-factor, the recombinant α-amylase was expressed and excreted out of the cells which was tested by SDS-PAGE. The recombinant α-amylase exhibited maximal activity at 90～100℃ and the optimum pH rangied from 4.0 to 4.5 that was 0.5 less than the optimum pH in the recombinat Pichia pastoris. Compared with the recombinat Pichia, the recombinat Hansenula polymorpha had more advantage such as simplifying the process of selection of the transformant, making it easier to get high-copies and more convenient for use in fermentation industry. |
| Keywords: | Pyrococcus furiosus |
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