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Evaluation of the potential for transformation of toxic compounds by Gordonia paraffinivorans and Gordonia sihwensis isolated from composting

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Stefania Pegorin Braga
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI)
Defense date:
Examining board members:
Renata Castiglioni Pascon; Welington Luiz de Araujo; Julio Cezar Franco de Oliveira; Marcela Pellegrini Peçanha
Advisor: Renata Castiglioni Pascon

Bioremediation is an efficient methodology on recovery of polluted environments. It acts employing microorganisms capable of degrading toxic compounds into less aggressive substances. Among the microorganisms used in bioremediation, the genus Gordonia has stood out due to its great metabolic versatility, which allows the use of complex toxic substrates as energy source, eliminating them from the environment. Sequencing and analysis of the genomes of two isolates of G. paraffinivorans (MTZ041 and 052) and an isolate of G. sihwensis (MTZ096) from composting, demonstrated that these microorganisms possess the genetic factors that confer the ability to degrade hydrocarbons, synthetic and natural rubber. To prove the capacity of growth of the isolates in these carbon sources, growth tests were carried out containing an oxide reduction indicator (2,6 DCPIP), kinetics growth, chemical analysis (gas chromatography, CG), qPCR and scanning electron microscopy (SEM). The three isolates used in this study were able to utilize hydrocarbons as sole carbon source. The results of the chromatographic assays show that G. paraffinivorans MTZ041 consumed 28.92% of the hexadecane present in the medium, against 69.80% consumed by G. sihwensis MTZ096 over 168 hours. The growth curves also showed that the biomass accumulated by G. sihwensis MTZ096 is 3.3 times higher than the biomass of G. paraffinivorans MTZ041 for the hexadecane treatment, resulting in 3.68 mg/ml and 1.11 mg/ml, respectively. As expected by these observations, the presence of hexadecane as carbon source in the medium led to an increased expression of the genes responsible for the degradation of hydrocarbons. For the CYP 153 System, genes FER, CYP 153 and FER RED had an increased expression of 750, 232 e 110 times, respectively. As for the Alkane Hydroxylase System, genes alk B, rub A3, rub A4, rub B and alk U had an increased expression of 248, 1576, 1296, 6,6 e 20 times, respectively. These results confirm previous data from the literature that Gordonia isolates have a great potential for biotransformation of hydrocarbons. Regarding assimilation of synthetic rubber and natural, in silico analysis demonstrated that only the isolates from the species G. paraffinivorans have the Lcp gene (latex Clearing Protein) responsible for the first step of the assimilation of these substrates. The growth curves in natural and synthetic rubber as carbon source were conducted over 11 weeks and demonstrate the ability of these substrates to support the robust growth of G. paraffinivorans . During 80 days, it was possible to evidence the presence of bacilli and the formation of biofilms on natural and synthetic rubber by SEM. This is the first report in the literature of G. paraffinivorans as a rubber degrader and the third species of the genus Gordonia with this ability. Given the complexity of this substrate and the shortage of micro-organisms with this capacity the description and characterization of this isolate is of great originality and biotechnological relevance. (AU)

FAPESP's process: 16/07360-0 - Evaluation of the transformation potential of toxic compounds by isolates Gordonia paraffinivorans and Gordonia sihwensis deriving from composting
Grantee:Stefania Pegorin Braga
Support Opportunities: Scholarships in Brazil - Master