• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1320-1324
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• 2006

The pancreatic and neutrophil elastases are associated with several illnesses including lung and vascular diseases, various cancers, and pancreatitis. The development of a potent and specific inhibitor to the elastases could lead to new therapies. In this study, an agar diffusion method was modified to include a substrate-dye conjugate (Elastin-Congo red) as a substrate of elastase and an indicator of elastase inhibitory activity. The Elastin-Congo red agar plates consisted of 0.1 % Elastin-Congo red and 2.5% agar. The elastase and elastase inhibitors were simultaneously loaded into wells, ultimately resulting in halo formations in which the halo diameter decreased as the concentration of elastase inhibitor increased. The concentration of elastase inhibitor in the samples, therefore, was inversely proportional to the halo diameters. This simplified method provided an excellent correlation with the standard microplate technique, which uses a chromogenic substrate. The concentration of elastase inhibitor obtained from the culture supernatant of a recombinant elastase inhibitor produced by the yeast Pichia pastoris was easily determined. This study has established a simple modified and inexpensive agar diffusion method that is potentially useful for the identification, quantification, and screening of new elastase inhibitors.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.2
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• pp.121-127
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• 1994

A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

• Journal of the Korea Organic Resources Recycling Association
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• v.2 no.2
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• pp.31-37
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• 1994

For a microbiological study of composting process, screening and assay method for biopolymer degrading enzymes and microorganisms were developed and for the study of the possibility of composting in anaerobic state, distribution of sulfate reducing bacteria which plays a final role in anaerobic degradation was investigated. Substrates used for the development of assay methods for biopolymer degradation are ${\beta}-glucan$, xylan, dextran, CMC(carboxy methly cellulose), casein, and collagen. These substrates were made insoluble by a cross-linking agent and linked with dye to make chromogenic substrates. ${\beta}-glucan$ and xylan substrates could substitute congo-red method for screening of polymer degrading microorganisms without damaging the colonies. Sulfate reducing bacteria contained in the sample sludge showed preference to lactic acid, propionic acid, butyric acid and formic acid and could use acetic acid and valeric acid.