Department for Biotechnology and Microbiology, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorfer Str. 1a, A-1060 Wien, Austria.
AMB Express. 2014 May 29;4:34. doi: 10.1186/s13568-014-0034-y. eCollection 2014.
We overexpressed the err1 gene in the Trichoderma reesei wild-type and in the cellulase hyperproducing, carbon catabolite derepressed strain Rut-C30 in order to investigate the possibility of producing erythritol with T. reesei. Two different promoters were used for err1 overexpression in both strains, a constitutive (the native pyruvat kinase (pki) promoter) and an inducible one (the native β-xylosidase (bxl1) promoter). The derived recombinant strains were precharacterized by analysis of err1 transcript formation on D-xylose and xylan. Based on this, one strain of each type was chosen for further investigation for erythritol production in shake flasks and in bioreactor experiments. For the latter, we used wheat straw pretreated by an alkaline organosolve process as lignocellulosic substrate. Shake flask experiments on D-xylose showed increased erythritol formation for both, the wild-type and the Rut-C30 overexpression strain compared to their respective parental strain. Bioreactor cultivations on wheat straw did not increase erythritol formation in the wild-type overexpression strain. However, err1 overexpression in Rut-C30 led to a clearly higher erythritol formation on wheat straw.
我们在里氏木霉野生型和纤维素酶高产、碳分解代谢阻遏解除的 Rut-C30 菌株中过表达 err1 基因,以研究用里氏木霉生产赤藓糖醇的可能性。在这两种菌株中,使用了两种不同的启动子进行 err1 的过表达,一种是组成型的(天然丙酮酸激酶(pki)启动子),另一种是诱导型的(天然β-木糖苷酶(bxl1)启动子)。通过分析 D-木糖和木聚糖上 err1 转录物的形成,对衍生的重组菌株进行了预表征。在此基础上,选择每种类型的一个菌株进行进一步研究,以在摇瓶和生物反应器实验中生产赤藓糖醇。对于后者,我们使用经过碱性有机溶解预处理的小麦秸秆作为木质纤维素底物。在 D-木糖上进行的摇瓶实验表明,与各自的亲本菌株相比,野生型和 Rut-C30 过表达菌株的赤藓糖醇产量都有所增加。在小麦秸秆上进行的生物反应器培养并没有增加野生型过表达菌株中的赤藓糖醇产量。然而,在 Rut-C30 中过表达 err1 导致在小麦秸秆上赤藓糖醇的形成明显增加。