National Research Institute of Brewing, Kagamiyama, Higashi-Hiroshima, Japan.
J Biosci Bioeng. 2013 Apr;115(4):394-9. doi: 10.1016/j.jbiosc.2012.10.025. Epub 2012 Nov 29.
The heterologous expression of the laccase gene from Trametes versicolor and Gaeumannomyces graminis was evaluated in the yeasts Pichia pastoris and Cryptococcus sp. S-2. The expression levels of both laccase genes in Cryptococcus sp. S-2 were considerably higher than those in P. pastoris. The codon usage of Cryptococcus sp. S-2 as well as the GC content were similar to those of T. versicolor and G. graminis. These results suggest that using a host with a similar codon usage for the expressed gene may improve protein expression. The use of Cryptococcus sp. S-2 as a host may be advantageous for the heterologous expression of genes with high GC content. Moreover, this yeast provides the same advantages as P. pastoris for the production of recombinant proteins, such as growth on minimal medium, capacity for high-density growth during fermentation, and capability for post-translational modifications. Therefore, we propose that Cryptococcus sp. S-2 be used as an expression host to improve enzyme production levels when other hosts have not yielded good results.
评估了来自彩绒革盖菌和禾顶囊壳菌的漆酶基因在毕赤酵母和隐球菌 S-2 中的异源表达。在隐球菌 S-2 中，两种漆酶基因的表达水平均显著高于毕赤酵母。隐球菌 S-2 的密码子使用和 GC 含量与彩绒革盖菌和禾顶囊壳菌相似。这些结果表明，使用密码子使用与表达基因相似的宿主可能会提高蛋白质表达水平。使用隐球菌 S-2 作为宿主可能有利于高 GC 含量基因的异源表达。此外，与毕赤酵母相比，该酵母在生产重组蛋白方面具有相同的优势，如在基础培养基上生长、发酵过程中高密度生长的能力以及翻译后修饰的能力。因此，当其他宿主未产生良好结果时，我们建议使用隐球菌 S-2 作为表达宿主以提高酶的产量水平。
