Phosphate released from phytic acid can be used as a measure of phytase activity. However, most of the phosphate estimation methods have not examined the interference or interaction of phytic acid in the assay. In this article, we report the kinetics and influence of unreduced phytic acid on phosphate estimation by three of the often-used methods for phytase estimation, the AOAC, Cooper-Gowing, and Fiske-Subbarow methods. Our results show that the AOAC method is most suitable to estimate the phytase activity in the presence of phytate in the medium. In the Fiske and Subbarow method, we noticed that the time factor plays a role in the interference of the phytic acid; especially the readings taken during the second hour of incubation are influenced by the presence of phytic acid. The method of Cooper and Gowing is labor-intensive and is prone to give error values at higher concentrations.
The indiscriminate usage of antibiotics has created a major problem in the form of antibiotic resistance. Even though new antimicrobial drug discovery programs have been in place from the last two decades, still we are unsuccessful in identifying novel molecules that have a potential to become new therapeutic agents for the treatment of microbial infections. A major problem in most screening studies is the requirement of high-throughput techniques. Given this, we present here an enzyme-based robust method for screening antimicrobial agent's active against Escherichia coli. This method is based upon the ability of the intracellular innate enzyme to cleave o-nitrophenyl β-d-galactopyranoside (non-chromogenic) to o-nitrophenolate (ONP) (chromogenic) upon the membrane damage or disruption. In comparison with the other currently available methods, we believe that our method provides an opportunity for real-time monitoring of the antimicrobial agents action by measuring the ONP generation in a user-friendly manner. Even though this method can be applied to other strain, our experience shows that one has to be careful especially when the pigments or metabolites present in the bacteria have the same wavelength absorbance.
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