Manipulating filamentous fungus chemical phenotypes by growth on nutritional arrays.
Methods for manipulating and fermenting microorganisms in multi-well plates offer unlimited possibilities for high-throughput parallel experimentation. Furthermore, bar-coded data tracking and downstream processing with modern liquid handling equipment reduce handling errors and are able to format microbial products for autosampler-equipped analytical instruments, e.g., HPLCs, mass spectrometers, and plate readers. An integrated system for high-throughput culturing of filamentous fungi replicating strains across many fermentation parameters, called nutritional arrays, was developed. It takes advantage of this equipment while addressing the age-old dilemma of how to manipulate fungal phenotypes to express a more complete spectrum of their secondary metabolites. Growth of any given strain in a well-designed nutritional array increases the chances of detecting a biologically active metabolite while reducing the manpower and materials needed for preparing individual fermentations and extracts. Fungi fermented in nutritional arrays are directly processed in a semi-automated fashion and the extracts prepared for bioassays and analytical chemistry. The necessary equipment, custom tools, and protocols to grow fungi in nutritional arrays are described along with examples of bioactive secondary metabolites discovered using this system.
Fundación MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Armilla, Granada, Spain. jose.tormo@medinaandalucia.es
José R
JR
Tormo
Francisco J
FJ
Asensio
Gerald F
GF
Bills
D046508:Culture Techniques / Q000295:instrumentation*; D005285:Fermentation; D005658:Fungi / Q000737:chemistry* / Q000254:growth & development* / Q000378:metabolism; D046228:Microarray Analysis / Q000295:instrumentation*; D050296:Microbial Viability; D010641:Phenotype