Microbial processes analytical methods

Although not within the scope of this chapter, it is worth noting that the PAT initiative also involves the development of rapid microbiological methods [46], the word rapid in this context being the analogue to process analytical for chemical and physical measurements. The scope of microbial testing within this initiative includes... 

The proof of activity of a biological pesticide is typically evaluated by a standardized bioassay except in the case of microbial metabolites where the major active ingredient(s) may be measured by analytical methods. Biological activity measurements, besides serving as a parameter for quality control, are an essential tool in the product development and optimization process. It is important to define the assay procedure in order to compare production batches and experimental formulations. These assays are typically used for product release or may be designed to assess specific aspects of product activity such as mobility in soils, colonization on leaf surface, etc. 

Other Applications. Fermentation-based processes have also been developed for the mass production of certain materials. Most notably, these processes have focused on the bulk production of alternative fuels, particularly ethanol and other solvents, and biogas, which is primarily methane. Processes employing Clostridia bacteria, for example, are useful in the production of acetone and 1-butanol, two extremely important organic solvents. In addition, as the applied science and technology behind fermentation becomes more developed, other areas of science will use the ability to tailor microbial processes for the purposes of analytical and test procedures and production methods. 

Use of an integrated system incorporating CCC separation, PDA detector, and LC-MS proved to be a valuable tool in the rapid identification of known compounds from microbial extracts.6 This collection of analytical data has enabled us to make exploratory use of advanced data analysis methods to enhance the identification process. For example, from the UV absorbance maxima and molecular weight for the active compound(s) present in a fraction, a list of potential structural matches from a natural products database (e.g., Berdy Bioactive Natural Products Database, Dictionary of Natural Products by Chapman and Hall, etc.) can be generated. Subsequently, the identity of metabolite(s) was ascertained by acquiring a proton nuclear magnetic resonance ( H-NMR) spectrum. 

Future cultivation methods will resemble existing methods of microbial and virus culture. Ill-defined medium components and cells will be replaced to enhance reproducibflity in production. For bacterial and ex vivo cultivated virus, analytical advances will make monitoring the environment and nutritional status of the culture more ubiquitous. However, the major changes will be in novel product types — single-molecule subunit antigens, viras-hke particles, monoclonal antibodies, and gene-therapy vaccines, each of which will incorporate novel processes. 

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