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Applied and Environmental Microbiology, June 2004, p. 3329-3337, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3329-3337.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Enzyme Production-Based Approach for Determining the Functions of Microorganisms within a Community

Kohei Nakamura,^* Shin Haruta, Huong Lan Nguyen, Masaharu Ishii, and Yasuo Igarashi

Department of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan

Received 7 November 2003/ Accepted 26 February 2004

The functions of specific microorganisms in a microbial community were investigated during the composting process. Cerasibacillus quisquiliarum strain BLx^T and Bacillus thermoamylovorans strain BTa were isolated and characterized in our previous studies based on their dominance in the composting system. Strain BLx^T 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 BLx^T. Zymograph analysis demonstrated that the pattern of active gelatinase bands from strain BLx^T was similar to that from the compost. Gelatinases from both BLx^T and compost were partially purified and compared. Homologous N-terminal amino acid sequences were found in one of the gelatinases from strain BLx^T and that of compost. These results indicate strain BLx^T 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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* Corresponding author. Mailing address: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan. Phone: 81 3 5841 5142. Fax: 81 3 5841 5272. E-mail: k-nakamura{at}mbj.nifty.com.

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Applied and Environmental Microbiology, June 2004, p. 3329-3337, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3329-3337.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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