Summary: A method is described for the specific hydrolysis of amino acids from humic substances by using the proteolytic enzyme "Pronase E". The amino acids are analyzed by a fluorescence detector after precolumn derivatization and HPLC-separation. The pattern and the concentration of amino acids after the enzymatic hydrolysis are set up for humic substances of different origin. Due to the results the enzymatic hydrolysis is suitable for the characterization of humic substances with a nitrogen content higher than 3%. As shown in a kinetic study of the hydrolysis of a fulvic acid the influence of humic substances on the enzyme activity increases in case of lower nitrogen content. Due to the specificity of the proteolytic enzyme the operationally defined method is limited as shown by a protein of a known amino acid content. The amino acids leucine, valine, tyrosine and histidine are released to the highest extent. The determined total extent for the hydrolysis of Insulin B is 26.34% and for Casein 38.37%. By comparison of the enzymatic hydrolysis with the classic acid hydrolysis the advantages of the biochemical method are the simple experimental handling and the mild reaction conditions. Amino acids which are very unstable like glutamine and asparagine can only be analyzed by the enzymatic hydrolysis.
Summary: Microbial biodegradation of organic substances takes place during drinking water treatment, but also in the distribution net, if the drinking water still contains biodegradable organic substances. This phenomenon is called regrowth. The regrowth potential of a drinking water is high, when for instance ozonation is used as the last step of treatment. It was proved before, that ozonation increases the biodegradability of humic substances, which are the main fraction of organic carbon in drinking water. In this work the objective was to check, if chlorination has a similar effect on humic substances. Using the method developed by Werner of measuring the regrowth potential of a water it could be shown by dilution series with chlorinated and unchlorinated humic substances that the substrate quality of these organics is increased by chlorination. The better substrate quality is seen in the chlorinated solutions, which might contain also low molecular weight organic compounds, but also in the fulvic acid fraction after XAD-enrichment, which removes most of the low molecular weight organic compounds. The chlorination creates in the fulvic acid solutions a shift of molecular size to smaller molecules and higher polar substances, which might be the reasons for the better biodegradability. From these results it can be concluded, that chlorination produces substances, which are more easily biodegradable. But this will not produce regrowth problems as long as there is free chlorine present as a disinfectant. In contrast, when the chlorine demand is very high and no free chlorine is left, this might produce high colony counts in the distribution net.
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