Regulation of two distinct alcohol oxidase promoters in the methylotrophic yeast Pichia methanolica
Regulation of two distinct alcohol oxidase promoters in the methylotrophic yeast Pichia methanolica
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DOI：
10.1002/yea.1334
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发表时间：
2006-01-15
期刊：
影响因子：
2.6
通讯作者：
Tomizuka, N
中科院分区：
文献类型：
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作者：
Nakagawa, T;Inagaki, A;Tomizuka, N
本研究以酿酒酵母酸性磷酸酶(AP)基因(ScPHO5)为报告基因，在启动子检测系统中对两种甲醇毕赤酵母乙醇氧化酶(AOD)启动子P-MOD1和P-MOD2进行了研究。由P-MOD1和P-MOD2启动子驱动的外源基因表达在转录水平上受到碳源的严格调控。P-MOD1不仅能被甲醇诱导，也能被甘油诱导。P-MOD2只被甲醇诱导，但不受添加甘油的抑制，这表明P-MOD2的调控方式与其他AOD-基因启动子不同。另一方面，甲醇和氧气水平影响P-MOD_1和P_MOD_2介导的基因表达。P-MOD1在低甲醇浓度下表达最好，而P-MOD2在高甲醇和高氧气浓度下优势表达。基于这些结果，P-MOD2和P-MOD1都应该是控制异源基因在甲氧基苯丙胺中表达的有用工具。特别是，在同一宿主细胞和同一烧瓶中使用P-MOD1和P-MOD2，应该可以不同地控制两种异源蛋白质的产生阶段。版权所有(C)2006 John Wiley&Sons，Ltd.
In this study, two Pichia methanolica alcohol oxidase (AOD) promoters, P-MOD1, and P-MOD2, were evaluated in a promoter assay system utilizing the acid phosphatase (AP) gene from Saccharomyces cerevisiae (ScPHO5) as a reporter. Heterologous gene expression driven by the P-MOD1 and P-MOD2 promoters was found to be strong and tightly regulated by carbon source at the transcriptional level. P-MOD1 was induced not only by methanol but also by glycerol. P-MOD2 was induced only by methanol, although it was not repressed on the addition of glycerol to a methanol medium, suggesting that P-MOD2 is regulated in a manner distinct from that of other AOD-gene promoters. On the other hand, methanol and oxygen level-influenced gene expression mediated by P-MOD1 and P-MOD2. P-MOD1 expression was optimal at low methanol concentrations, whereas P-MOD2 was predominantly expressed at high methanol and high oxygen concentrations. Based on these results, both P-MOD2 and P-MOD1 should be useful tools for controlling heterologous gene expression in P. methanolica. In particular, it should be possible to differentially control the production phases of two heterologous proteins, using P-MOD1 and P-MOD2 in the same host cell and in the same flask. Copyright (c) 2006 John Wiley & Sons, Ltd.