Analysis of FR901379 Biosynthetic Genes in <i>Coleophoma empetri</i> by Clustered Regularly Interspaced Short Palindromic Repeats/Cas9-Based Genomic Manipulation

Wei, Teng-Yun; Zheng, Yufeng; Wan, Miyang; Yang, Songbai; Tang, Jiawei; Wu, Yuanjie; Li, Jiyang; Chen, Shaoxin

doi:10.1021/acschembio.2c00250

Cited by 6 publications

(15 citation statements)

References 44 publications

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“…The production of FR901379 was abolished completely in the mcfJ deletion mutant. It is consistent with the phenomenon observed in C. empetri SIPI1284, deleting Cehyp (homologous gene of mcfJ ) broke the production of FR901379 [ 26 ]. The transcriptional levels of the FR901379 biosynthetic pathway, including the genes in the large mcf cluster and the separate O -sulfonation cluster, were significantly downregulated following mcfJ disruption (Fig.…”

Section: Resultssupporting

confidence: 88%

“…In transformants MEFC09-HF-7 and MEFC09-HF-8, the titer of WF11899B was significantly increased and much higher than that of FR901379. This might be due to the imbalance between McfF and McfH caused by the uncertainty of integration sites and copies of expression cassettes [ 26 , 28 ]. In other transformants, the titers of FR901379 were improved significantly compared to the parental strains MEFC09-H-6 and MEFC09, while the proportions of WF11899B and WF11899C were both reduced (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, the biosynthetic pathway of FR901379 in C. empetri has been elucidated by gene deletion, enzymatic assays, and heterologous refactoring [ 26 , 27 ]. The involved biosynthetic genes are distributed in two separate biosynthetic gene clusters (BGCs), the core biosynthetic gene cluster mcf and the O -sulfonation gene cluster (Fig.…”

Section: Introductionmentioning

confidence: 99%

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Improving the production of the micafungin precursor FR901379 in an industrial production strain

et al. 2023

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Background Micafungin is an echinocandin-type antifungal agent used for the clinical treatment of invasive fungal infections. It is semisynthesized from the sulfonated lipohexapeptide FR901379, a nonribosomal peptide produced by the filamentous fungus Coleophoma empetri. However, the low fermentation efficiency of FR901379 increases the cost of micafungin production and hinders its widespread clinical application. Results Here, a highly efficient FR901379-producing strain was constructed via systems metabolic engineering in C. empetri MEFC09. First, the biosynthesis pathway of FR901379 was optimized by overexpressing the rate-limiting enzymes cytochrome P450 McfF and McfH, which successfully eliminated the accumulation of unwanted byproducts and increased the production of FR901379. Then, the functions of putative self-resistance genes encoding β-1,3-glucan synthase were evaluated in vivo. The deletion of CEfks1 affected growth and resulted in more spherical cells. Additionally, the transcriptional activator McfJ for the regulation of FR901379 biosynthesis was identified and applied in metabolic engineering. Overexpressing mcfJ markedly increased the production of FR901379 from 0.3 g/L to 1.3 g/L. Finally, the engineered strain coexpressing mcfJ, mcfF, and mcfH was constructed for additive effects, and the FR901379 titer reached 4.0 g/L under fed-batch conditions in a 5 L bioreactor. Conclusions This study represents a significant improvement for the production of FR901379 and provides guidance for the establishment of efficient fungal cell factories for other echinocandins.

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Section: Resultssupporting

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Improving the production of the micafungin precursor FR901379 in an industrial production strain

et al. 2023

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“…This sulfated echinocandin is a precursor of the partial chemical synthesis of the echinocandin, micafungin, commercially sold as Mycamine and Fungiguard [ 103 ]. Recently, the BGCs involved in the biosynthesis of FR901379 (16) were analyzed and an efficient clustered regularly interspaced short palindromic repeats/Cas9-based gene editing tool for the industrial production strain C. empetri SIPI1284 was established [ 104 ].…”

Section: Bioactive Fungal Secondary Metabolites and Derivatives Succe...mentioning

confidence: 99%

Overview of Bioactive Fungal Secondary Metabolites: Cytotoxic and Antimicrobial Compounds

et al. 2022

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Microorganisms are known as important sources of natural compounds that have been studied and applied for different purposes in distinct areas. Specifically, in the pharmaceutical area, fungi have been explored mainly as sources of antibiotics, antiviral, anti-inflammatory, enzyme inhibitors, hypercholesteremic, antineoplastic/antitumor, immunomodulators, and immunosuppressants agents. However, historically, the high demand for new antimicrobial and antitumor agents has not been sufficiently attended by the drug discovery process, highlighting the relevance of intensifying studies to reach sustainable employment of the huge world biodiversity, including the microorganisms. Therefore, this review describes the main approaches and tools applied in the search for bioactive secondary metabolites, as well as presents several examples of compounds produced by different fungi species with proven pharmacological effects and additional examples of fungal cytotoxic and antimicrobial molecules. The review does not cover all fungal secondary metabolites already described; however, it presents some reports that can be useful at any phase of the drug discovery process, mainly for pharmaceutical applications.

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“…Notably, significant progress has been made in the establishment of transformation systems mediated by protoplasts and Agrobacterium for G. lozoyensis , A. pachycristatuc , and C. empetri . Moreover, in all of these strains, gene‐editing systems have been adeptly established, including homologous recombination and/or CRISPR/Cas9 (Table 1 ; Cacho et al., 2012 ; Chen et al., 2013 ; Men et al., 2021 ; Wei et al., 2020 ; Wei et al., 2022 ). In the field of fermentation, the augmentation of production yields of pneumocandin B 0 , echinocandin B, and FR901379 has been meticulously pursued through the optimization of carbon–nitrogen sources and cultivation conditions.…”

Section: Introductionmentioning

confidence: 99%

Insight into advances for the biosynthetic progress of fermented echinocandins of antifungals

Jiang,

Luo,

Wang

et al. 2023

Microbial Biotechnology

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Invasive fungal infections have increased remarkably, which have become unprecedented concern to human health. However, the effectiveness of current antifungal drugs is limited due to drug resistance and toxic side‐effects. It is urgently required to establish the effective biosynthetic strategy for developing novel and safe antifungal molecules economically. Echinocandins become a promising option as a mainstay family of antifungals, due to specifically targeting the fungal specific cell wall. To date, three kinds of echinocandins for caspofungin, anidulafungin, and micafungin, which derived from pneumocandin B0, echinocandin B, and FR901379, are commercially available in clinic and have shown potential in managing invasive fungal infections in a cost‐effective manner. However, current echinocandins‐derived precursors all are produced by environmental fungal isolates with long fermentation cycle and low yields, which challenge the production efficacy of these precursors in industry. Therefore, understanding their biosynthetic machinery is of great importance for improving antifungal titres and creating new echinocandins‐derived products. With the development of genome‐wide sequencing and establishment of gene‐editing technology, there are a growing number of reports on echinocandins‐derived products and their biosynthetic gene clusters. This review briefly summarizes the discovery and development history of echinocandins, compares their structural characteristics and biosynthetic processes, and sums up existed strategies for improving their production. Moreover, the genomic analysis of related biosynthetic gene clusters of echinocandins is discussed, highlighting the similarities and differences among the clusters. Last, the biosynthetic processes of echinocandins are compared, focusing on the activation and attachment of side‐chains and the formation of the hexapeptide core. This review aims to provide insights into the development and production of new echinocandin drugs by modifying the structure of echinocandin‐derived precursors and/or optimizing the fermentation processes; and achieve a new microbial chassis for efficient production of echinocandins in heterologous hosts.

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Analysis of FR901379 Biosynthetic Genes in Coleophoma empetri by Clustered Regularly Interspaced Short Palindromic Repeats/Cas9-Based Genomic Manipulation

Cited by 6 publications

References 44 publications

Improving the production of the micafungin precursor FR901379 in an industrial production strain

Improving the production of the micafungin precursor FR901379 in an industrial production strain

Overview of Bioactive Fungal Secondary Metabolites: Cytotoxic and Antimicrobial Compounds

Insight into advances for the biosynthetic progress of fermented echinocandins of antifungals

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