The initial aim of this project was to uncover novel cellulolytic organisms or enzymes from the diverse microbial source, activated sludge. Two isolation methods were used; either directly inoculating the sludge material onto filter paper as a carbon source, or using the Evolver� technology as an enrichment device. In both cases, as expected, cellulase activity was evident, however attributing this activity to one species was difficult in either case. This highlighted the complex interrelationships that existed between the many microorganisms present as the cellulosic carbon sources were degraded. In one instance, a Cellvibrio sp. was isolated. This genus of bacteria is known to possess both types of cellulase activity (exo- and endo- acting) and was therefore likely to contribute to the degradation of the cellulose. However, the isolate, once purified, did not display significant cellulolytic ability as compared to the unpurified consortium of microorganisms. Therefore, in each case, microorganisms responsible for the cellulolytic activity were not uncovered. It was suspected that the microorganisms responsible for some of the cellulolytic activity were protists. During the isolation of microorganisms, an Aeromonas sp. bearing the novel phenotype (for this genus) of CMCase activity was isolated. This activity was at first suspected to contribute to the degradation of the filter paper that was seen during isolation. However, tests with the pure isolate suggested that the Aeromonas sp. CMCase was not used for cellulose catabolism. Ironically, the enzyme may instead function in the production of a cellulose-like exopolysaccharide by the bacterium. Part of a cellulose synthase operon was found in the genome of the Aeromonas sp. isolate, including a gene coding for an endoglucanase that gives a predicted molecular weight enzyme similar to the 39 kDa CMCase purified from the bacterium. The CMCase enzyme, operating as part of of a synthetic operon is expected to be important in terms of the biofilm forming ability of this Aeromonas strain. Such capabilities of the bacterium were investigated here, including observing motility behaviour of the organism on agar surfaces. Studying the biofilm forming ability of this genus in general will be important in understanding how the fish and human pathogens persist in aquatic environments
| Identifer | oai:union.ndltd.org:ADTP/235185 |
| Date | January 2007 |
| Creators | Clinton, Brook, brook.clinton@csiro.au |
| Publisher | RMIT University. Applied Sciences |
| Source Sets | Australiasian Digital Theses Program |
| Language | English |
| Detected Language | English |
| Rights | http://www.rmit.edu.au/help/disclaimer, Copyright Brook Clinton |
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