气候变化领域集成服务门户(CCPortal): The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi

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DOI	10.1073/pnas.2025689118
	The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi
	Gabriel R.; Thieme N.; Liu Q.; Li F.; Kohler L.T.; Harth S.; Jecmenica M.; Ramamurthy M.; Gorman J.; Simmons B.A.; McCluskey K.; Baker S.E.; Tian C.; Schuerg T.; Singer S.W.; Fleißner A.; Benz J.P.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:26
英文摘要	Carbohydrate active enzymes (CAZymes) are vital for the lignocellulose-based biorefinery. The development of hypersecreting fungal protein production hosts is therefore a major aim for both academia and industry. However, despite advances in our understanding of their regulation, the number of promising candidate genes for targeted strain engineering remains limited. Here, we resequenced the genome of the classical hypersecreting Neurospora crassa mutant exo-1 and identified the causative point of mutation to reside in the F-box protein–encoding gene, NCU09899. The corresponding deletion strain displayed amylase and invertase activities exceeding those of the carbon catabolite derepressed strain Δcre-1, while glucose repression was still mostly functional in Δexo-1. Surprisingly, RNA sequencing revealed that while plant cell wall degradation genes are broadly misexpressed in Δexo-1, only a small fraction of CAZyme genes and sugar transporters are up-regulated, indicating that EXO-1 affects specific regulatory factors. Aiming to elucidate the underlying mechanism of enzyme hypersecretion, we found the high secretion of amylases and invertase in Δexo-1 to be completely dependent on the transcriptional regulator COL-26. Furthermore, misregulation of COL-26, CRE-1, and cellular carbon and nitrogen metabolism was confirmed by proteomics. Finally, we successfully transferred the hypersecretion trait of the exo-1 disruption by reverse engineering into the industrially deployed fungus Myceliophthora thermophila using CRISPR-Cas9. Our identification of an important F-box protein demonstrates the strength of classical mutants combined with next-generation sequencing to uncover unanticipated candidates for engineering. These data contribute to a more complete understanding of CAZyme regulation and will facilitate targeted engineering of hypersecretion in further organisms of interest. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	CAZymes gene regulation; Enzyme hypersecretion; F-box proteins; Fungal biotechnology; Neurospora crassa
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/238499
作者单位	Deconstruction Division, Joint BioEnergy Institute, Emeryville, CA 94608, United States; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Emeryville, CA 94608, United States; Institut für Genetik, Technische Universität Braunschweig, Braunschweig, 38106, Germany; Technical University of Munich School of Life Sciences Weihenstephan, Technical University of Munich, Freising, 85354, Germany; Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China; Fungal Genetics Stock Center, Department of Plant Pathology, Kansas State University, Manhattan, KS 66506, United States; Functional and Systems Biology, Environmental Molecular Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, United States; Braunschweig Integrated Centre of Systems Biology, Technische Universität Braunschweig, Braunschweig, 38106, Germany; Institute for Advanced Study, Technical Universit...
推荐引用方式 GB/T 7714	Gabriel R.,Thieme N.,Liu Q.,et al. The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi[J],2021,118(26).
APA	Gabriel R..,Thieme N..,Liu Q..,Li F..,Kohler L.T..,...&Benz J.P..(2021).The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi.Proceedings of the National Academy of Sciences of the United States of America,118(26).
MLA	Gabriel R.,et al."The F-box protein gene exo-1 is a target for reverse engineering enzyme hypersecretion in filamentous fungi".Proceedings of the National Academy of Sciences of the United States of America 118.26(2021).