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鉴定一种参与调控[具体物种名称未给出]中植物生物质降解酶产生的新型转录因子TP05746

Identification of a Novel Transcription Factor TP05746 Involved in Regulating the Production of Plant-Biomass-Degrading Enzymes in .

作者信息

Zhang Ting, Liao Lu-Sheng, Li Cheng-Xi, Liao Gui-Yan, Lin Xiong, Luo Xue-Mei, Zhao Shuai, Feng Jia-Xun

机构信息

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, Nanning, China.

出版信息

Front Microbiol. 2019 Dec 13;10:2875. doi: 10.3389/fmicb.2019.02875. eCollection 2019.

Abstract

Limited information on transcription factor (TF)-mediated regulation exists for most filamentous fungi, specifically for regulation of the production of plant-biomass-degrading enzymes (PBDEs). The filamentous fungus, , can secrete integrative cellulolytic and amylolytic enzymes, suggesting a promising application in biotechnology. In the present study, the regulatory roles of a Zn2Cys6 protein, TP05746, were investigated in through the use of biochemical, microbiological and omics techniques. Deletion of the gene in led to a 149.6% increase in soluble-starch-degrading enzyme (SSDE) production at day one of soluble starch induction but an approximately 30% decrease at days 2 to 4 compared with the parental strain Δ. In contrast, the . mutant Δ exhibited a 136.8-240.0% increase in raw-starch-degrading enzyme (RSDE) production, as well as a 90.3 to 519.1% increase in cellulase and xylanase production following induction by culturing on wheat bran plus Avicel, relative to that exhibited by Δ. Additionally, the mutant Δ exhibited accelerated mycelial growth at the early stage of cultivation and decreased conidiation. Transcriptomic profiling and real-time quantitative reverse transcription-PCR (RT-qPCR) analyses revealed that TP05746 dynamically regulated the expression of genes encoding major PBDEs and their regulatory genes, as well as fungal development-regulated genes. Furthermore, binding experiments confirmed that TP05746 bound to the promoter regions of the genes described above. These results will contribute to our understanding of the regulatory mechanism of PBDE genes and provide a promising target for genetic engineering for improved PBDE production in filamentous fungi.

摘要

对于大多数丝状真菌,尤其是关于植物生物质降解酶(PBDEs)产生的调控,转录因子(TF)介导的调控信息有限。丝状真菌 能够分泌综合的纤维素分解酶和淀粉分解酶,这表明其在生物技术领域具有广阔的应用前景。在本研究中,通过生化、微生物学和组学技术,研究了一种Zn2Cys6蛋白TP05746在 中的调控作用。与亲本菌株Δ相比,在 中缺失基因 导致在可溶性淀粉诱导的第一天,可溶性淀粉降解酶(SSDE)产量增加149.6%,但在第2至4天下降约30%。相比之下, 突变体Δ在以麦麸加微晶纤维素培养诱导后,原生淀粉降解酶(RSDE)产量增加136.8 - 240.0%,纤维素酶和木聚糖酶产量增加90.3至519.1%。此外,突变体Δ在培养早期菌丝体生长加快,分生孢子形成减少。转录组分析和实时定量逆转录PCR(RT-qPCR)分析表明,TP05746动态调节编码主要PBDEs及其调控基因以及真菌发育调控基因的表达。此外, 结合实验证实TP05746与上述基因的启动子区域结合。这些结果将有助于我们理解PBDE基因的调控机制,并为丝状真菌中提高PBDE产量的基因工程提供一个有前景的靶点。

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