Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0

Pneumocandin B0 (PB0) is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate (Cancidas®). However, the low titer of PB0 results in fermentation and purification costs during caspofungin production, limiting its widesp...

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Main Authors: Xinyi Zhang, Shu Cheng, Jing Yang, Li Lu, Zixin Deng, Guangkai Bian, Tiangang Liu
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2025-06-01
Series:Synthetic and Systems Biotechnology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405805X24001601
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author Xinyi Zhang
Shu Cheng
Jing Yang
Li Lu
Zixin Deng
Guangkai Bian
Tiangang Liu
author_facet Xinyi Zhang
Shu Cheng
Jing Yang
Li Lu
Zixin Deng
Guangkai Bian
Tiangang Liu
author_sort Xinyi Zhang
collection DOAJ
description Pneumocandin B0 (PB0) is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate (Cancidas®). However, the low titer of PB0 results in fermentation and purification costs during caspofungin production, limiting its widespread clinical application. Here, we engineered an efficient PB0-producing strain of G. lozoyensis by systems metabolic engineering strategies, including multi-omics analysis and multilevel metabolic engineering. We overexpressed four rate-limiting enzymes: thioesterase GLHYD, two cytochrome P450s GLP450s, and chorismate synthase GLCS; knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E; and overexpressed the global transcriptional activator GLHYP. As a result, the PB0 titer increased by 108.7 % to 2.63 g/L at the shake-flask level through combinatorial strategies. Our study provides valuable insights into achieving high-level production of PB0 and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.
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institution Kabale University
issn 2405-805X
language English
publishDate 2025-06-01
publisher KeAi Communications Co., Ltd.
record_format Article
series Synthetic and Systems Biotechnology
spelling doaj-art-4852c6ff608e4119bbf0482bd29755f32025-08-20T03:47:20ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2025-06-0110238139010.1016/j.synbio.2024.12.008Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0Xinyi Zhang0Shu Cheng1Jing Yang2Li Lu3Zixin Deng4Guangkai Bian5Tiangang Liu6Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, China; Center of Materials Synthetic Biology, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 518055, Shenzhen, ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, China; State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200030, Shanghai, ChinaCenter of Materials Synthetic Biology, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 518055, Shenzhen, China; Corresponding author.Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, China; State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 200030, Shanghai, China; Corresponding author. Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and School of Pharmaceutical Sciences, Wuhan University, 430072, Wuhan, China.Pneumocandin B0 (PB0) is a lipohexapeptide synthesized by Glarea lozoyensis and serves as the precursor for the widely used antifungal drug caspofungin acetate (Cancidas®). However, the low titer of PB0 results in fermentation and purification costs during caspofungin production, limiting its widespread clinical application. Here, we engineered an efficient PB0-producing strain of G. lozoyensis by systems metabolic engineering strategies, including multi-omics analysis and multilevel metabolic engineering. We overexpressed four rate-limiting enzymes: thioesterase GLHYD, two cytochrome P450s GLP450s, and chorismate synthase GLCS; knocked out two competing pathways responsible for producing 6-methylsalicylic acid and pyranidine E; and overexpressed the global transcriptional activator GLHYP. As a result, the PB0 titer increased by 108.7 % to 2.63 g/L at the shake-flask level through combinatorial strategies. Our study provides valuable insights into achieving high-level production of PB0 and offers general guidance for developing efficient fungal cell factories to produce polyketide synthase-non-ribosomal peptide synthetase hybrid metabolites.http://www.sciencedirect.com/science/article/pii/S2405805X24001601Antifungal drugEchinocandinsPneumocandin B0Multi-omicsSystems metabolic engineering
spellingShingle Xinyi Zhang
Shu Cheng
Jing Yang
Li Lu
Zixin Deng
Guangkai Bian
Tiangang Liu
Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
Synthetic and Systems Biotechnology
Antifungal drug
Echinocandins
Pneumocandin B0
Multi-omics
Systems metabolic engineering
title Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
title_full Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
title_fullStr Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
title_full_unstemmed Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
title_short Metabolic engineering of Glarea lozoyensis for high-level production of pneumocandin B0
title_sort metabolic engineering of glarea lozoyensis for high level production of pneumocandin b0
topic Antifungal drug
Echinocandins
Pneumocandin B0
Multi-omics
Systems metabolic engineering
url http://www.sciencedirect.com/science/article/pii/S2405805X24001601
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AT lilu metabolicengineeringofglarealozoyensisforhighlevelproductionofpneumocandinb0
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