Department of Cellular Biology, Institute of Biological Sciences, University of Brasilia, Brasilia-DF 70.910-900, Brazil.
Enzyme Microb Technol. 2011 Jan 5;48(1):19-26. doi: 10.1016/j.enzmictec.2010.08.007. Epub 2010 Sep 21.
Time-course expression profiles of one xylanase and eight cellulase encoding genes, as well as of two transcription factor encoding genes of Humicola grisea var. thermoidea were established in different culture media pHs and carbon sources (glucose and sugarcane bagasse). Quantitative real-time RT-PCR analysis revealed a remarkable and parallel increase in mRNA accumulation for cbh1.1, cbh1.2, egl1, egl2, egl3, bgl4 and xyn1 at alkaline pH and with sugarcane bagasse employed as the sole carbon source. Glucose utilization led to a higher creA mRNA accumulation compared to the other genes. A distinct pattern was observed for egl4, whose mRNA preferably accumulated in acidic conditions. The transcriptional profile data combined with the analysis of the in vitro binding of PacC and CreA transcription factors to the promoters support the CreA-mediated carbon repression and the PacC-related pH regulation of H. grisea cellulase and xylanase encoding genes. Moreover, EMSA analyses suggest a role for CreA on pacC transcriptional regulation. These data will be useful to H. grisea hydrolytic enzymes production improvement, as well as to the design of optimized promoters aiming industrial heterologous proteins production.
建立了嗜热毁丝霉变种在不同培养介质 pH 值和碳源(葡萄糖和甘蔗渣)下一个木聚糖酶和八个纤维素酶编码基因以及两个转录因子编码基因的时程表达谱。定量实时 RT-PCR 分析显示,在碱性 pH 值和以甘蔗渣作为唯一碳源时,cbh1.1、cbh1.2、egl1、egl2、egl3、bgl4 和 xyn1 的 mRNA 积累显著且平行增加。与其他基因相比,葡萄糖利用导致 creA mRNA 积累更高。egl4 的 mRNA 则表现出明显的偏好酸性条件下积累的模式。转录谱数据结合 PacC 和 CreA 转录因子与启动子体外结合的分析表明,CreA 介导的碳抑制和 PacC 相关的 pH 调节嗜热毁丝霉纤维素酶和木聚糖酶编码基因。此外,EMSA 分析表明 CreA 在 pacC 转录调控中起作用。这些数据将有助于提高嗜热毁丝霉水解酶的生产,以及设计用于工业异源蛋白生产的优化启动子。