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AMB Express
2016 Mar 01;61:17. doi: 10.1186/s13568-016-0188-x.
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Cellulolytic and proteolytic ability of bacteria isolated from gastrointestinal tract and composting of a hippopotamus.
da Cruz Ramos GF
,
Ramos PL
,
Passarini MRZ
,
Vieira Silveira MA
,
Okamoto DN
,
de Oliveira LCG
,
Zezzo LV
,
Marem A
,
Santos Rocha RC
,
da Cruz JB
,
Juliano L
,
de Vasconcellos SP
.
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The bioprospection for cellulase and protease producers is a promise strategy for the discovery of potential biocatalysts for use in hydrolysis of lignocellulosic materials as well as proteic residues. These enzymes can increment and turn viable the production of second generation ethanol from different and alternative sources. In this context, the goal of this study was the investigation of cellulolytic and proteolytic abilities of bacteria isolated from the gastrointestinal tract of a hippopotamus as well as from its composting process. It is important to highlight that hippopotamus gastrointestinal samples were a non-typical sources of efficient hydrolytic bacteria with potential for application in biotechnological industries, like biofuel production. Looking for this, a total of 159 bacteria were isolated, which were submitted to qualitative and quantitative enzymatic assays. Proteolytic analyzes were conducted through the evaluation of fluorescent probes. Qualitative assays for cellulolytic abilities revealed 70 positive hits. After quantitative analyzes, 44 % of these positive hits were selected, but five (5) strains showed cellulolytic activity up to 11,8 FPU/mL. Regarding to proteolytic activities, six (6) strains showed activity above 10 %, which overpassed results described in the literature. Molecular analyzes based on the identification of 16S rDNA, revealed that all the selected bacterial isolates were affiliated to Bacillus genus. In summary, these results strongly indicate that the isolated bacteria from a hippopotamus can be a potential source of interesting biocatalysts with cellulolytic and proteolytic activities, with relevance for industrial applications.
Fig. 1.
a Illustration of the process to construct of a composting pile at São Paulo Zoo Park Foundation; b Illustration of technicians sampling material from the composting pile; c Adopted instrument that was applied to access all the layers that form the composting pile (Bitencourt et al. 2010); d Ready sample for microbiological analyzes
Fig. 2. Abundance of bacterial isolates in CFU (colony-forming unit) at different culture media, when incubated at two temperatures: 30 and 45 °C
Fig. 3. Representation using dispersion curves of enzymatic indexes obtained by 70 selected bacteria
Fig. 4. Cellulolytic activity represented as Filter Paper Units (FPU/mL) of five (5) selected strains, under pH 4.8 and pH 7.4, compared to T. reesei (positive control)
Fig. 5. Graphic representation about the proteolytic activity of five (5) bacteria, under pH 7.0 and six (6) isolates under pH 9.0 (cut off 10 %)
Fig. 6. Phylogenetic analysis based on partial 16S rRNA sequences of the FPZSP isolates and related Bacillus species. The tree reconstruction was generated from Neighbour-Joining method, following p distance model. Bootstrap values (1000 replicate runs, %) >70 % are listed. GenBank accession numbers are listed after species names. E. coli was used as outgroup
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