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Chromobacterium Sp. From The Tropics: Detection And Diversity Of Phytase Activity

Phytases are a group of enzymes that catalyze phytic acid hydrolysis with release of phosphorus (P). The ability of Chromobacterium sp. to produce phytase was detected in 115 out of 118 candidate bacteria isolated from different Brazilian biomas. This is the first report revealing the genus Chromobacterium as phytase producer.

isolates were divided into three classes of phytase production, according to diameter of halos: low (6-9.9 mm), medium (10-15.9 mm) and high (16-18 mm). Association between classes of phytase production and origin of the isolates was not observed.    Phytase activity has been widely found among microorganisms such as bacteria, yeasts and filamentous fungi.
The Proteobacteria phylum include most phytase-producing bacteria although the β-proteobacteria class, which the genus Chromobacterium belongs to, bears only two phytase-producer genera, Burkholderia and Acidovora (7). Although there was no evidence for phytase genes in the annotated genome of C.
violaceum ATCC 12472 T type species (2), our study showed in vitro phytase production in Chromobacterium sp. isolates from several Brazilian ecosystems. To our knowledge, this is the first report of phytase-producing isolates in the genus

Chromobacterium.
Some Chromobacterium isolates that had been classified as low producers in the qualitative screen exhibited high enzymatic activity. This apparently conflicting result may have arisen due to technical reasons intrinsic to each methodology.
Such type of problem has been reported in the determination of antimicrobial activity where differences in the sensitivity profile of microorganisms were observed depending on whether liquid or solid media had been used; this has been a concern, for example, of the Clinical and Laboratory Standards Institute (3). In the quantitative assay, bacteria were grown in liquid medium, thus phytate was more readily available allowing easier degradation. Another possibility is that some isolates secreted lower amounts of enzyme in solid media. It is known that in liquid media bacteria express different factors compared to growth in solid media; one example is the capsular polysaccharide which may interfere with antibacterial activity of drugs such as lysostaphin (9). It is also possible that different phytases vary in their diffusion rates through agar. In Chromobacterium sp. from the tropics any case, we consider that the qualitative method should be used only as an initial screening and should not be used as a precise comparative method for phytase production.
Variability in phytase production was a remarkable feature of the isolates and diversity has been observed for other characteristics in Chromobacterium (11). Variation in genome sizes within the same species is common in bacteria. According to Parkhill and Thomson (17) the genome of Escherichia coli can vary over 1 Mb among natural isolates. There is indication that much of this diversity is due to the presence of bacteriophages (14). We presume that the same phenomenon may have happened in Chromobacterium allowing for different gene diversity among natural isolates which would explain the phytase diversity observed in this study. Therefore, we suggest that the C. violaceum type species, even though it was the one chosen for genome sequencing (2), is not a representative of the isolates found in nature. The type species may in fact lack a phytase gene, differently from the several isolates analyzed here. Since phytases are enzymes grouped mainly for their structure and catalytic properties rather than sequence homology (8, 7) individual gene analysis is not a very straightforward approach and genomic studies may be more appropriate.
The biotechnological potential of C. violaceum has been explored and includes bioremediation, production of PHAs