Development of a robust microtiter plate-based assay method for assessment of bioactivity.
A microtiter plate-based assay was developed for the quantitative monitoring of bioactive compound production in Streptomyces hygroscopicus fermentation samples. The method reported demonstrates the successful application of the theories of disk diffusion based methods of bioactivity assessment, to a microtiter assay for high throughput analysis. The assay method facilitates the generation of the dose-response curve of test organisms (Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae) to a bioactive compound. Using this dose-response curve, the method facilitates definition of three distinct Minimum Inhibitory Concentration (MIC) values for use in the characterisation of the bioactive attributes of a sample. The assay uses established standard procedures to facilitate adaptation of the assay for use with a wider range of test microorganisms. Errors due to the assumption of a linear relationship between turbidity and biomass concentration are also reduced, due to incorporation of a step to convert turbidity to biomass concentration, for use in the calculation of bioactivity.
Casey
J T
JT
Bioprocess Engineering Research Group, National Institute for Cellular Biotechnology and School of Biotechnology, Dublin City University, Dublin 9, Ireland.
O'Cleirigh
C
C
Walsh
P K
PK
O'Shea
D G
DG
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000187:drug effects*; D005285:Fermentation; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009391:Nephelometry and Turbidimetry; D012441:Saccharomyces cerevisiae / Q000187:drug effects*; D013302:Streptomyces / Q000378:metabolism*
D000900:Anti-Bacterial Agents