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  • 1
    Publication Date: 2023-03-14
    Keywords: Biomass; Biomass, standard error; Experiment day; pH; pH, standard deviation; Species; Strain; Time in hours
    Type: Dataset
    Format: text/tab-separated-values, 288 data points
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  • 2
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    PANGAEA
    In:  Supplement to: Kramer, Annemarie; Beck, Hans Christian; Kumar, Abhishek; Kristensen, Lars Peter; Imhoff, Johannes F; Labes, Antje (2015): Proteomic Analysis of Anti-Cancerous Scopularide Production by a Marine Microascus brevicaulis Strain and Its UV Mutant. PLoS ONE, 10(10), e0140047, https://doi.org/10.1371/journal.pone.0140047
    Publication Date: 2023-01-13
    Description: The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularide A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing besides higher production levels faster growth and differences in pellet formation. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of this fungus and its mutant. For this purpose, an optimised protein extraction protocol was established. Here, we show the first proteome study of a marine fungus. In total, 4759 proteins were identified. The central metabolic pathway of LF580 could be mapped by using KEGG pathway analysis and GO annotation. Using iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to a limited nutrient availability in wild type strain due to a strong pellet formation. This information can be applied to optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum.
    Type: Dataset
    Format: application/zip, 2 datasets
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  • 3
    Publication Date: 2023-01-13
    Keywords: Accession number; Category; Comment; Description; Enzyme code; Gene Ontology term
    Type: Dataset
    Format: text/tab-separated-values, 1828 data points
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  • 4
    Publication Date: 2019-09-23
    Description: Strains originally affiliated to the genera Scopulariopsis and Microascus were compared regarding the scopularide production in order to investigate their ability to produce the cyclodepsipeptides and select the best suited candidate for subsequent optimisation processes. Phylogenetic calculations using available sequences of the genera Scopulariopsis and Microascus revealed that most of the sequences clustered within two closely related groups, comprising mainly Scopulariopsis/Microascus brevicaulis and Microascus sp., respectively. Interestingly, high yields of scopularide A were exhibited by three strains belonging to S./M. brevicaulis, while lower titres were observed for two strains of Microascus sp. Close phylogenetic distances within and between the two groups supported the proposed combination of both genera into one holomorph group. Short phylogenetic distances did not allow a clear affiliation at the species level on the basis of ribosomal DNA sequences, especially for Microascus sp. strains. Additionally, several sequences originating from strains assigned to Scopulariopsis exhibited a polyphyletic nature. The production pattern is in accordance with the phylogenetic position of the strains and significant production of scopularide B could only be observed for the S./M. brevicaulis strain LF580. Thus, the phylogenetic position marks the biotechnologically interesting strains and matters in optimisation strategies. In conclusion, the ability of all five strains to produce at least one of the scopularides suggests a distribution of the responsible gene cluster within the holomorph group. Setting the focus on the production of the cyclodepsipeptides, strain LF580 represents the best candidate for further strain and process optimisation
    Type: Article , PeerReviewed , info:eu-repo/semantics/article
    Format: text
    Format: text
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  • 5
    Publication Date: 2019-09-23
    Description: Natural compounds from marine fungi are an excellent source for the discovery and development of new drug leads. The distinct activity profiles of the two cyclodepsipeptides scopularide A and B against cancer cell lines set their marine producer strain Scopulariopsis brevicaulis LF580 into the focus of the EU project MARINE FUNGI. One of the main goals was the development of a sustainable biotechnological production process for these compounds. The secondary metabolite production of strain LF580 was optimized by random mutagenesis employing UV radiation. For a fast and reliable detection of the intracellular secondary metabolite production level, a miniaturized bioactivity-independent screening method was developed, as the random mutagenesis yielded a large number of mutants to be analysed quantitatively and none of the existing hyphenated bioassay-dependent screening systems could be applied. The method includes decreased cultivation volume, a fast extraction procedure as well as an optimized LC-MS analysis. We show that deviation could be specifically reduced at each step of the process: The measuring deviation during the analysis could be minimized to 5% and technical deviation occurring in the downstream part to 10–15%. Biological variation during the cultivation process still has the major influence on the overall variation. However, the approach led to a 10-fold reduction of time and similar effects on costs and effort compared to standard reference screening methods. The method was applied to screen the UV-mutants library of Scopulariopsis brevicaulis LF580. For validation purposes, the occurring variations in the miniaturized scale were compared to those in the classical Erlenmeyer flask scale. This proof of concept was performed using the wild type strain and 23 randomly selected mutant strains. One specific mutant strain with an enhanced production behavior could be obtained.
    Type: Article , PeerReviewed , info:eu-repo/semantics/article
    Format: text
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  • 6
    Publication Date: 2019-09-23
    Description: Background: Marine organisms produce many novel compounds with useful biological activity, but are currently underexploited. Considerable research has been invested in the study of compounds from marine bacteria, and several groups have now recognised that marine fungi also produce an interesting range of compounds. During product discovery, these compounds are often produced only in non-agitated culture conditions, which are unfortunately not well suited for scaling up. A marine isolate of Scopulariopsis brevicaulis, strain LF580, produces the cyclodepsipeptide scopularide A, which has previously only been produced in non-agitated cultivation. Results: Scopulariopsis brevicaulis LF580 produced scopularide A when grown in batch and fed-batch submerged cultures. Scopularide A was extracted primarily from the biomass, with approximately 7% being extractable from the culture supernatant. By increasing the biomass density of the cultivations, we were able to increase the volumetric production of the cultures, but it was important to avoid nitrogen limitation. Specific production also increased with increasing biomass density, leading to improvements in volumetric production up to 29-fold, compared with previous, non-agitated cultivations. Cell densities up to 36 g L-1 were achieved in 1 to 10 L bioreactors. Production of scopularide A was optimised in complex medium, but was also possible in a completely defined medium. Conclusions: Scopularide A production has been transferred from a non-agitated to a stirred tank bioreactor environment with an approximately 6-fold increase in specific and 29-fold increase in volumetric production. Production of scopularide A in stirred tank bioreactors demonstrates that marine fungal compounds can be suitable for scalable production, even with the native production organism.
    Type: Article , PeerReviewed , info:eu-repo/semantics/article
    Format: text
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  • 7
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    Unknown
    In:  [Poster] In: ELRIG 2014 Research and Innovation - Impact into Cancer, 11.03.2014, Telford, UK .
    Publication Date: 2019-09-23
    Type: Conference or Workshop Item , NonPeerReviewed
    Format: text
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  • 8
    Publication Date: 2019-09-23
    Description: The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant. For this purpose, an optimised protein extraction protocol was established. In total, 4759 proteins were identified. The central metabolic pathway of strain LF580 was mapped using the KEGG pathway analysis and GO annotation. Employing iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum
    Type: Article , PeerReviewed
    Format: text
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  • 9
    Publication Date: 2019-09-23
    Description: Filamentous fungi are well known for their capability of producing antibiotic natural products. Recent studies have demonstrated the potential of antimicrobials with vast chemodiversity from marine fungi. Development of such natural products into lead compounds requires sustainable supply. Marine biotechnology can significantly contribute to the production of new antibiotics at various levels of the process chain including discovery, production, downstream processing, and lead development. However, the number of biotechnological processes described for large-scale production from marine fungi is far from the sum of the newly-discovered natural antibiotics. Methods and technologies applied in marine fungal biotechnology largely derive from analogous terrestrial processes and rarely reflect the specific demands of the marine fungi. The current developments in metabolic engineering and marine microbiology are not yet transferred into processes, but offer numerous options for improvement of production processes and establishment of new process chains. This review summarises the current state in biotechnological production of marine fungal antibiotics and points out the enormous potential of biotechnology in all stages of the discovery-to-development pipeline. At the same time, the literature survey reveals that more biotechnology transfer and method developments are needed for a sustainable and innovative production of marine fungal antibiotics.
    Type: Article , PeerReviewed
    Format: text
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  • 10
    Publication Date: 2021-03-04
    Description: An unusual polyketide with a new carbon skeleton, lindgomycin (1), and the recently described ascosetin (2) were extracted from mycelia and culture broth of different Lindgomycetaceae strains, which were isolated from a sponge of the Kiel Fjord in the Baltic Sea (Germany) and from the Antarctic. Their structures were established by spectroscopic means. In the new polyketide, two distinct domains, a bicyclic hydrocarbon and a tetramic acid, are connected by a bridging carbonyl. The tetramic acid substructure of compound 1 was proved to possess a unique 5-benzylpyrrolidine-2,4-dione unit. The combination of 5-benzylpyrrolidine-2,4-dione of compound 1 in its tetramic acid half and 3-methylbut-3-enoic acid pendant in its decalin half allow the assignment of a new carbon skeleton. The new compound 1 and ascosetin showed antibiotic activities with IC50 value of 5.1 (±0.2) µM and 3.2 (±0.4) μM, respectively, against methicillin-resistant Staphylococcus aureus.
    Type: Article , PeerReviewed
    Format: text
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