Wang Zhengran, Zhao Shanshan, Zhang Kai, Lin Congyu, Ru Xin, Yang Qian
School of Life Science and Technology, Harbin Institute of Technology, No. 2 Yikuang Street, Nangang District, Harbin, 150080, People's Republic of China.
Synth Syst Biotechnol. 2022 Jul 16;7(4):1084-1094. doi: 10.1016/j.synbio.2022.07.002. eCollection 2022 Dec.
Cytochalasans, with diverse structures and pharmacological activities, are a class of compounds containing isoindolinone moieties fused to the tricyclic or tetracyclic ring system. Chaetoglobosin A (cheA), mainly produced by , is the most abundant cytochalasan. However, limited understanding of transcriptional regulation of morphological development and cheA biosynthesis in has hindered cheA application in agriculture and biomedical field. This study examined the regulatory role of gene in . had significant effect on secondary metabolites production in , similar to that reported in other filamentous fungi. Inactivation of caused an obvious decrease in cheA production from 51.32 to 19.76 mg/L under dark conditions. In contrast, overexpression resulted in a dramatic increase in cheA production, reaching 206.59 mg/L under light conditions, which was higher than that noted under dark condition. The RT-qPCR results confirmed that as a light responsive regulator, positively regulated cheA biosynthesis by controlling the expression of core genes of the cheA biosynthetic gene cluster and other relevant regulators. Electrophoretic mobility shift assays proved that directly regulated , and , and indirectly regulated . Moreover, had a significant effect on the regulation of asexual spores production. When compared with wild-type , silenced and overexpression mutants presented remarkable differences in sporulation, irrespective of light or dark condition. Besides, expression was speculated to negatively regulate spore formation. These findings illustrated the regulatory mechanism of a hypothetical global regulator, in , suggesting its potential application in industrial-scale cheA biosynthesis.
细胞松弛素具有多样的结构和药理活性,是一类含有与三环或四环环系统稠合的异吲哚啉酮部分的化合物。主要由[未提及的来源]产生的Chaetoglobosin A(cheA)是最丰富的细胞松弛素。然而,对[未提及的对象]中形态发育和cheA生物合成的转录调控了解有限,这阻碍了cheA在农业和生物医学领域的应用。本研究考察了[未提及的基因]在[未提及的对象]中的调控作用。[未提及的基因]对[未提及的对象]中次级代谢产物的产生有显著影响,这与其他丝状真菌的报道类似。在黑暗条件下,[未提及的基因]失活导致cheA产量从51.32毫克/升明显降至19.76毫克/升。相反,[未提及的基因]过表达导致cheA产量显著增加,在光照条件下达到206.59毫克/升,高于黑暗条件下的产量。RT-qPCR结果证实,[未提及的基因]作为光响应调节因子,通过控制cheA生物合成基因簇的核心基因和其他相关调节因子的表达,正向调控cheA的生物合成。电泳迁移率变动分析证明,[未提及的基因]直接调控[未提及的基因]、[未提及的基因]和[未提及的基因],并间接调控[未提及的基因]。此外,[未提及的基因]对无性孢子产生的调控有显著影响。与野生型[未提及的对象]相比,[未提及的基因]沉默和[未提及的基因]过表达突变体在孢子形成方面表现出显著差异,无论光照或黑暗条件如何。此外,推测[未提及的基因]表达负向调节孢子形成。这些发现阐明了一种假设的全局调节因子[未提及的基因]在[未提及的对象]中的调控机制,表明其在工业规模cheA生物合成中的潜在应用。