Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No.12 South Zhongguancun St., Haidian District, Beijing, 100081, China.
Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No. 2 West Yuanmingyuan Road, Haidian District, Beijing, 100193, China.
Microb Cell Fact. 2023 Feb 25;22(1):38. doi: 10.1186/s12934-023-02046-w.
Trichoderma reesei (T. reesei) is well-known for its excellent ability to secret a large quantity of cellulase. However, unlike the endogenous proteins, little is known about the molecular mechanisms governing heterologous protein production. Herein, we focused on the integration loci and the secretory pathway, and investigated their combinatorial effects on heterologous gene expression in T. reesei using a glucose oxidase from Aspergillus niger as a model protein. Integration in the cel3c locus was more efficient than the cbh1 locus in expressing the AnGOx by increasing the transcription of AnGOx in the early stage. In addition, we discovered that interruption of the cel3c locus has an additional effect by increasing the expression of the secretory pathway component genes. Accordingly, overexpressing three secretory pathway component genes, that were snc1, sso2, and rho3, increased AnGOx expression in the cbh1 transformant but not in the cel3c transformant.
里氏木霉（T. reesei）以其能够大量分泌纤维素酶的能力而闻名。然而，与内源性蛋白不同，人们对调控异源蛋白生产的分子机制知之甚少。在此，我们专注于整合位点和分泌途径，并使用黑曲霉葡萄糖氧化酶作为模型蛋白，研究它们对里氏木霉中异源基因表达的组合效应。在 cel3c 基因座的整合比在 cbh1 基因座的整合更有效地表达 AnGOx，通过增加 AnGOx 在早期的转录。此外，我们发现中断 cel3c 基因座通过增加分泌途径组成基因的表达具有额外的作用。因此，过表达三个分泌途径组成基因，即 snc1、sso2 和 rho3，增加了 cbh1 转化体中 AnGOx 的表达，但在 cel3c 转化体中没有增加。
