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青霉属 NCIM 1366 中纤维素酶诱导的早期细胞事件和潜在调控因子。

Early cellular events and potential regulators of cellulase induction in Penicillium janthinellum NCIM 1366.

机构信息

Biofuels and Biorefineries Section, Microbial Processes and Technology Division, CSIR- National Institute for Interdisciplinary Science and Technology, Industrial Estate P.O., Pappanamcode, Thiruvananthapuram, Kerala, 695019, India.

Academy of Scientific and Innovative Research (AcSIR), Ghaziabad- 201002, India.

出版信息

Sci Rep. 2023 Mar 28;13(1):5057. doi: 10.1038/s41598-023-32340-x.

Abstract

Cellulase production by fungi is tightly regulated in response to environmental cues, and understanding this mechanism is a key pre-requisite in the efforts to improve cellulase secretion. Based on UniProt descriptions of secreted Carbohydrate Active enZymes (CAZymes), 13 proteins of the cellulase hyper-producer Penicillium janthinellum NCIM 1366 (PJ-1366) were annotated as cellulases- 4 cellobiohydrolases (CBH), 7 endoglucanases (EG) and 2 beta glucosidases (BGL). Cellulase, xylanase, BGL and peroxidase activities were higher for cultures grown on a combination of cellulose and wheat bran, while EG was stimulated by disaccharides. Docking studies indicated that the most abundant BGL- Bgl2- has different binding sites for the substrate cellobiose and the product glucose, which helps to alleviate feedback inhibition, probably accounting for the low level of glucose tolerance exhibited. Out of the 758 transcription factors (TFs) differentially expressed on cellulose induction, 13 TFs were identified whose binding site frequencies on the promoter regions of the cellulases positively correlated with their abundance in the secretome. Further, correlation analysis of the transcriptional response of these regulators and TF-binding sites on their promoters indicated that cellulase expression is possibly preceded by up-regulation of 12 TFs and down-regulation of 16 TFs, which cumulatively regulate transcription, translation, nutrient metabolism and stress response.

摘要

真菌细胞的产酶过程受到环境信号的严格调控,因此了解这一调控机制是提高纤维素酶分泌效率的关键前提。根据 UniProt 对分泌性碳水化合物活性酶(CAZymes)的描述,纤维素高产菌青霉(Penicillium janthinellum NCIM 1366)PJ-1366 中的 13 种蛋白被注释为纤维素酶,包括 4 种纤维二糖水解酶(CBH)、7 种内切葡聚糖酶(EG)和 2 种β葡萄糖苷酶(BGL)。在同时含有纤维素和麦麸的培养基中培养时,PJ-1366 的纤维素酶、木聚糖酶、BGL 和过氧化物酶活性更高,而 EG 则受到二糖的刺激。对接研究表明,含量最丰富的 BGL-Bgl2 具有不同的结合部位,分别用于结合底物纤维二糖和产物葡萄糖,这有助于缓解反馈抑制,可能是其表现出较低葡萄糖耐受性的原因。在纤维素诱导的差异表达的 758 个转录因子(TFs)中,鉴定出 13 个 TF 的结合位点频率与它们在分泌组中的丰度呈正相关,这些 TF 存在于纤维素诱导表达的 758 个转录因子中。此外,对这些调控因子的转录响应及其启动子上 TF 结合位点的相关性分析表明,纤维素酶的表达可能是在 12 个 TF 的上调和 16 个 TF 的下调之后发生的,这一系列调控作用共同调节转录、翻译、养分代谢和应激反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72f9/10050438/6352122857d1/41598_2023_32340_Fig1_HTML.jpg

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