Establishment of a Laboratory of Applied Microbiology in São Paulo Zoo: identification and isolation of microorganisms producing enzymes and its inhibitors of use in medical, veterinary and industrial areas

Abstract

The general objective of this project is to isolate, to know, to preserve and characterize microorganisms from the composting facility and water (from the lakes) of the São Paulo Zoological. Preliminary data show that traditional methods of isolation and preservation can be used for prospection of cultivable microorganisms from composting; however, it is estimated that most of the microorganisms that inhabit this material are not cultivable. Isolated microorganisms will be identified by mass spectrometry (MALDI/TOF-MS), a methodology that is being implemented in the laboratory of biophysics of INFAR/UNIFESP. The identifications will be validated by sequencing the small subunit of the ribosome (SSU rDNA). The microorganisms isolated from compost will be tested for production of enzymes of great industrial interest, such as celluloses, xylanases, ligninases, keratinase, amylase and protease, as well as the production of some inhibitors of these enzymes. At the same time, non-cultivable microbial diversity will be assessed by pyrosequencing of the total DNA extracted from this material at different stages of the maturation process of composting, using the sequencer 454/Roche installed at the Centre for Advanced Technologies in Genomics (CATG), Department of Biochemistry Institute of Chemistry, USP (IQUSP). In silico analysis of the sequences should generate a collection of predicted genes. These will be compared with the families of genes of enzymes of industrial interest, with the goal of finding new members of these gene families, homologous eventually present in non-isolatable organisms from composting. The expectation is that this research project generates new tools for biotechnological applications, a collection of isolated microorganisms, plus information on microbial genomes, all from an ecological niche not yet exploited, which represents a large gene pool for the scientific community. (AU)