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BIOTECHNOLOGY EXPLORATION OF GENE ALKB CLONED FROM LIBRARY METAGENOMICS ENCODING THE ENZYME ALKANE HYDROXYLASE

Grant number: 12/21913-1
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): June 01, 2013
Effective date (End): May 31, 2015
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Eliana Gertrudes de Macedo Lemos
Grantee:Silvana Pompeia Do Val de Moraes
Home Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

In Brazil there are approximately 35 thousand gas stations built in the 70s. With an average lifespan of 25 years for underground tanks, it is assumed that they are already committed. According to CETESB, the posts account for 63% of the contaminated areas in Sao Paulo, only 1 drop of lubricant oil pollutes 100 gallons of water. The Guarani aquifer, covering a total area of approximately 1.2 million Km - 839,800 km2 in the Center-West and South regions of Brazil -, stores around 46,000 km3 of water. The sustainable extraction considers the demand from 360 million people, with a per capita consumption of 300 l / inhab. day. The conserved volume could supply water to Brazil's population for 3,500 years. One of the large recharge areas of that aquifer is located in the north of São Paulo. The contamination of groundwater by leaks at gas stations is a growing concern in Brazil and oldest in the United States and Europe. Since the late 90's the oil struggle daily with problems resulting from leaks, spills and accidents during exploration, refining, transportation and storage operations of oil and its derivatives. Recent advances in molecular biology have extended our understanding of the metabolic processes related to microbial transformation of petroleum hydrocarbons. The physiological responses of microorganisms to the presence of hydrocarbons, including cell surface alterations and adaptive mechanisms for uptake and efflux of these substrates, have been characterized. New molecular techniques have enhanced our ability to investigate the dynamics of microbial communities in petroleum-impacted ecosystems. The alkB gene is the principal enzyme involved in the synthesis of alkane hydroxylase gene product alkB acts by a mechanism of its own direct reversal processing the removal of damage from methylation of DNA bases through a reaction of oxygen dependent, alpha-ketoglutarate and Fe (II). From a consortium of microorganisms resulting from soil contaminated with diesel fuel, was constructed a metagenomic library that belongs to the Laboratory of Biochemistry Plants and Microorganisms (LBMP) and having identified a gene encoding the enzyme alkB alkane hydroxylase. Given this result we saw the possibility of entering into the expression vector and sub cloning for being a completely new gene that had not been described This project proposes the use of strategically designed to investigate the potential molecular biotransformador alkB gene cloned in order to contribute to application in a future technology