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Enzyme Production-Based Approach for Determining the Functions of Microorganisms within a Community
Authors:Kohei Nakamura  Shin Haruta  Huong Lan Nguyen  Masaharu Ishii  and Yasuo Igarashi
Abstract:The functions of specific microorganisms in a microbial community were investigated during the composting process. Cerasibacillus quisquiliarum strain BLxT and Bacillus thermoamylovorans strain BTa were isolated and characterized in our previous studies based on their dominance in the composting system. Strain BLxT degrades gelatin, while strain BTa degrades starch. We hypothesized that these strains play roles in gelatinase and amylase production, respectively. The relationship between changes in the abundance ratios of each strain and those of each enzyme activity during the composting process was examined to address this hypothesis. The increase in gelatinase activity in the compost followed a dramatic increase in the abundance ratio of strain BLxT. Zymograph analysis demonstrated that the pattern of active gelatinase bands from strain BLxT was similar to that from the compost. Gelatinases from both BLxT and compost were partially purified and compared. Homologous N-terminal amino acid sequences were found in one of the gelatinases from strain BLxT and that of compost. These results indicate strain BLxT produces gelatinases during the composting process. Meanwhile, the increase in the abundance ratio of strain BTa was not concurrent with that of amylase activity in the compost. Moreover, the amylase activity pattern of strain BTa on the zymogram was different from that of the compost sample. These results imply that strain BTa may not produce amylases during the composting process. To our knowledge, this is the first report demonstrating that the function of a specific microorganism is directly linked to a function in the community, as determined by culture-independent and enzyme-level approaches.
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