The work undertaken explored the potential for discovery of new bioactive metabolites from soil fungi. The research developed selective mycological isolation techniques and maximised metabolite production from active selected fungi by application of the OSMAC approach and concept of hormesis. Novel active compounds were discovered and many other known compounds characterised.

The work undertaken explored the potential for discovery of new bioactive metabolites from soil fungi. The research developed selective mycological isolation techniques and maximised metabolite production from active selected fungi by application of the OSMAC approach and concept of hormesis. Novel active compounds were discovered and many other known compounds characterised.

>Magister Scientiae - MSc / The discovery of novel secondary metabolites has declined significantly in recent years whereas there is a rise in the number of multi-drug resistant pathogens and other types of diseases. The decline in natural product discovery was due to high rediscovery of already known compounds and the costs in developing natural products. As a result pharmaceutical companies lost interest in investing in natural product discovery. However, there is a renewed interest in marine sponge associated microorganisms as a rich and untapped source of secondary metabolites. The objective of this study was to design a matrix to investigate the extent to which the One Strain-Many Compounds (OSMAC) approach applies to a collection of marine sponge isolates harvested from two South African marine sponge samples. Terminal restriction fragment length polymorphisms (T-RFLP) analysis was used to investigate and ascertain the two marine sponges which hosted the highest microbial diversities to be used for further culture-dependent studies. The culture-dependent studies, using 33 media which included liquid enrichment, heat treatments and antibiotic treatments, resulted in 400 sponge isolates from the two marine sponges Isodictya compressa and Higginsia bidentifera. Using antibacterial overlay assays, 31 dereplicated isolates showed antibacterial activity. Bioactivities were also exhibited against E. coli 1699 which is genetically engineered for resistance against 52 antibiotics which implies that some of the bioactive compounds could be novel. The 16S rRNA gene sequences revealed that the microbial phyla isolated from the marine sponges belonged to Actinobacteria, Firmicutes and Proteobacteria (Alphaproteobacteria and Gammaproteobacteria).Thirty isolates were selected for an OSMAC-based matrix study, 17 of which showed noantibacterial activities in preliminary screening. The application of the OSMAC approach using co-culture and 36 culture conditions resulted in 6 isolates showing antibacterial activities, three of which did not show activities in preliminary screening. One of these, a Bacillus pumilus isolated from I. compressa displayed antibacterial activity against 5 indicator strains whereas in preliminary screening it had not shown activity. The results show that marine sponges can host novel microbial species which may produce novel bioactive compounds. The results also confirm that traditional methods employing a single culture condition restricts the expression of some biosynthetic pathways of microorganisms and as a result many metabolites have yet to be identified.

Orientador: Ian Castro-Gamboa / Resumo: A utilização de produtos naturais pela humanidade, como agentes terapêuticos, é descrita desde primórdios da antiguidade e representam um importante aliado no tratamento de doenças. Com isso a busca por novas ferramentas analíticas mais robustas, com abordagem metabolômica se faz necessária, de forma a otimizar e acelerar a busca por novas moléculas e substâncias com atividade biológica promissora. Nesse contexto, esse trabalho tem como alvo de estudo, os fungos Fusarium solani e Purpureocillium lilacinum isolados da rizosfera de Senna spectabilis. Essas espécies de microrganismos apresentam muitos estudos químicos relatados na literatura, o que influenciou o interesse em realizar os estudos de OSMAC (One Strain Many Compounds). Para, assim, verificar a variação metabólica que acontece com esses fungos em diferentes tipos de cultivo (variando meio de cultura e agitação) incluindo o co-cultivo. Para os extratos obtidos também foram realizados os ensaios biológicos citotóxico e antibacteriano. Foram utilizadas ferramentas analíticas do estado da arte como GC-MS (Cromatografia Gasosa acoplada a Espectrometria de Massas) e LC-MS (Cromatografia Líquida acoplada a Espectrometria de Massas) fazendo uso das ferramentas de bioinformática Molecular Networking (ou Rede Molecular) com a plataforma GNPS (Global Natural Product Social Molecular Networking) e o UNIFI (Sistema de informação científica – Waters Corporation). A espectrometria de massas (MS) foi uma grande aliada que ajudou a d... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The use of natural products by mankind, as therapeutic agents, has been described since ancient times and represents an important ally in the treatment of diseases. Thus, the search for new more robust analytical tools, with a metabolomic approach is necessary, in order to optimize and accelerate the search for new molecules and substances with promising biological activity. In this context, this study is aimed at studying Fusarium solani and Purpureocillium lilacinum fungi isolated from the rhizosphere of Senna spectabilis. These species of microorganisms have many chemical studies reported in the literature, which influenced the interest in carrying out the studies of OSMAC (One Strain Many Compounds). Thus, to verify the metabolic variation that happens with these fungi in different types of cultivation (varying culture medium and agitation) including co-culture. For the extracts obtained, cytotoxic and antibacterial biological tests were also performed. State-of-the-art analytical tools such as GCMS (Gas Chromatography coupled to Mass Spectrometry) and LC-MS (Liquid Chromatography coupled to Mass Spectrometry) were used alongside the bioinformatics tools Molecular Networking with the platform GNPS (Global Natural Product Social Molecular Networking) and UNIFI (Scientific Information System - Waters Corporation). Mass spectrometry (MS) was a great ally that helped to demonstrate the variation of the metabolites profile of these microorganisms in different cultures, in whic... (Complete abstract click electronic access below) / Doutor

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