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pH调节剂pac-3的缺失影响粗糙脉孢菌降解甘蔗渣过程中纤维素酶和木聚糖酶的活性。

Deletion of pH Regulator pac-3 Affects Cellulase and Xylanase Activity during Sugarcane Bagasse Degradation by Neurospora crassa.

作者信息

Campos Antoniêto Amanda Cristina, Ramos Pedersoli Wellington, Dos Santos Castro Lílian, da Silva Santos Rodrigo, Cruz Aline Helena da Silva, Nogueira Karoline Maria Vieira, Silva-Rocha Rafael, Rossi Antonio, Silva Roberto Nascimento

机构信息

Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

出版信息

PLoS One. 2017 Jan 20;12(1):e0169796. doi: 10.1371/journal.pone.0169796. eCollection 2017.

Abstract

Microorganisms play a vital role in bioethanol production whose usage as fuel energy is increasing worldwide. The filamentous fungus Neurospora crassa synthesize and secrete the major enzymes involved in plant cell wall deconstruction. The production of cellulases and hemicellulases is known to be affected by the environmental pH; however, the regulatory mechanisms of this process are still poorly understood. In this study, we investigated the role of the pH regulator PAC-3 in N. crassa during their growth on sugarcane bagasse at different pH conditions. Our data indicate that secretion of cellulolytic enzymes is reduced in the mutant Δpac-3 at alkaline pH, whereas xylanases are positively regulated by PAC-3 in acidic (pH 5.0), neutral (pH 7.0), and alkaline (pH 10.0) medium. Gene expression profiles, evaluated by real-time qPCR, revealed that genes encoding cellulases and hemicellulases are also subject to PAC-3 control. Moreover, deletion of pac-3 affects the expression of transcription factor-encoding genes. Together, the results suggest that the regulation of holocellulase genes by PAC-3 can occur as directly as in indirect manner. Our study helps improve the understanding of holocellulolytic performance in response to PAC-3 and should thereby contribute to the better use of N. crassa in the biotechnology industry.

摘要

微生物在生物乙醇生产中发挥着至关重要的作用,生物乙醇作为燃料能源在全球的使用量正在不断增加。丝状真菌粗糙脉孢菌能合成并分泌参与植物细胞壁解构的主要酶类。已知纤维素酶和半纤维素酶的产生受环境pH值影响;然而,这一过程的调控机制仍知之甚少。在本研究中,我们调查了pH调节剂PAC-3在不同pH条件下粗糙脉孢菌在甘蔗渣上生长过程中的作用。我们的数据表明,在碱性pH条件下,突变体Δpac-3中纤维素分解酶的分泌减少,而在酸性(pH 5.0)、中性(pH 7.0)和碱性(pH 10.0)培养基中,木聚糖酶受到PAC-3的正向调控。通过实时定量PCR评估的基因表达谱显示,编码纤维素酶和半纤维素酶的基因也受PAC-3调控。此外,pac-3的缺失会影响编码转录因子的基因的表达。总之,结果表明PAC-3对全纤维素酶基因的调控可能以直接或间接的方式发生。我们的研究有助于增进对PAC-3响应下全纤维素分解性能的理解,从而有助于在生物技术产业中更好地利用粗糙脉孢菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/337f/5249074/331a2bc9acc8/pone.0169796.g001.jpg

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