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聚乙二醇8000胁迫下里氏木霉中通过钙信号传导对纤维素酶产生的调控

Regulation of cellulase production via calcium signaling in Trichoderma reesei under PEG8000 stress.

作者信息

Liu Shuai, Quan Lin, Yang Mei, Wang Dan, Wang Yong-Zhong

机构信息

Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, 400030, China.

School of Chemistry and Chemical Engineering, Chongqing University, Chongqing, 400044, China.

出版信息

Appl Microbiol Biotechnol. 2024 Jan 26;108(1):178. doi: 10.1007/s00253-023-12901-w.

Abstract

In this study, the effect of polyethylene glycol 8000 (PEG8000) stress on cellulase biosynthesis in Trichoderma reesei CICC2626 via calcium signaling was investigated, and a plausible mechanism by which intracellular Ca regulates the transcription of cellulase genes was proposed. The results indicated that the total cellulase (filter paper-hydrolyzing activity [FPase]), endoglucanase (carboxymethyl cellulase activity [CMCase]), and β-glucosidase activities of the strain were 1.3-, 1.2-, and 1.3-fold higher than those of the control (no PEG8000 addition) at a final concentration of 1.5% (w/v) PEG8000. Moreover, the transcriptional levels of cellulase genes, protein concentrations, and biomass increased. With the synergistic use of commercial cellulase and T. reesei CICC2626 cellulase to hydrolyze alkali-pretreated rice straw, the released reducing sugar concentration reached 372.7 mg/g, and the cellulose content (22.7%, 0.32 g) was significantly lower than the initial content (62.5%, 1.88 g). Transcriptome data showed that 12 lignocellulose degradation-related genes were significantly upregulated in the presence of 1.5% PEG8000. Furthermore, the addition of Ca inhibitors and deletion of crz1 (calcineurin-responsive zinc finger 1-encoding gene, which is related to the calcium signaling pathway) demonstrated that calcium signaling plays a dominant role in PEG8000-induced cellulase genes overexpression. These results revealed a link between PEG8000 induction and calcium signaling transduction in T. reesei CICC2626. Moreover, this study also provides a novel inducer for enhanced cellulase production. KEY POINTS: • Cellulase biosynthesis in Trichoderma reesei could be enhanced by PEG8000 • PEG8000 could induce a cytosolic Ca burst in Trichoderma reesei • The activated calcium signaling was involved in cellulase biosynthesis.

摘要

本研究通过钙信号转导研究了聚乙二醇8000(PEG8000)胁迫对里氏木霉CICC2626中纤维素酶生物合成的影响,并提出了细胞内Ca调节纤维素酶基因转录的可能机制。结果表明,在终浓度为1.5%(w/v)的PEG8000条件下,该菌株的总纤维素酶(滤纸水解活性[FPase])、内切葡聚糖酶(羧甲基纤维素酶活性[CMCase])和β-葡萄糖苷酶活性分别比对照(未添加PEG8000)高1.3倍、1.2倍和1.3倍。此外,纤维素酶基因的转录水平、蛋白质浓度和生物量均有所增加。将商业纤维素酶与里氏木霉CICC2626纤维素酶协同用于水解碱预处理稻草,释放的还原糖浓度达到372.7 mg/g,纤维素含量(22.7%,0.32 g)显著低于初始含量(62.5%,1.88 g)。转录组数据显示,在1.5% PEG8000存在下,12个木质纤维素降解相关基因显著上调。此外,添加Ca抑制剂和缺失crz1(与钙信号通路相关的编码钙调神经磷酸酶反应性锌指1的基因)表明,钙信号在PEG8000诱导的纤维素酶基因过表达中起主导作用。这些结果揭示了里氏木霉CICC2626中PEG8000诱导与钙信号转导之间的联系。此外,本研究还为提高纤维素酶产量提供了一种新型诱导剂。要点:• PEG8000可增强里氏木霉中的纤维素酶生物合成 • PEG8000可诱导里氏木霉胞质Ca爆发 • 激活的钙信号参与纤维素酶生物合成

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e1c/10817842/8c468a10ac11/253_2023_12901_Fig1_HTML.jpg

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