Riassunto analitico
Aromatic compounds are among the most prevalent and persistent pollutants in the environment. Petroleum-contaminated soil and sediment commonly contain a mixture of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatics. Aromatics derived from industrial activities often have functional groups such as alkyls, halogens, and nitro groups. Biodegradation is a major mechanism of removal of organic pollutants from a contaminated site. Syntrophorhabdus aromaticivorans is an obligate anaerobic bacterium that can grow on isophthalate and lives in syntrophy with sulfate-reducing Desulfovibrio species. When growing S. aromaticivorans with isophthalate, four proteins could be identified in high abundance by means of proteome analysis. One of these proteins, called isophthaloyl-CoA decarboxylase (IPCD), catalyzes the decarboxylation of isophthaloyl-CoA to benzoyl-CoA under anaerobic conditions.. In this study the encoding genes of IPCD from S. aromaticivorans were successfully cloned into different expression vectors to enable heterologous production in E. coli or Thauera chlorobenzoica.
The following results have been achieved:
1) Transformation of the gene sequences coding for the isophthaloyl-CoA decarboxylase (IPCD) enzyme with pRSF-Duet1 vector by restriction cloning or fast cloning into Escherichia coli NEB5α.
2) Transformation of the gene sequences coding for the isophthaloyl-CoA decarboxylase (IPCD) enzyme with pIZ1016 and POT1 into Thauera Chlorobenzoica.
3) Transferring and enhancing the genetic system of Thauera chlorobenzoica 3CB-1 strain adopting protocols of Thauera aromatica.
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Abstract
Aromatic compounds are among the most prevalent and persistent pollutants in the environment. Petroleum-contaminated soil and sediment commonly contain a mixture of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatics. Aromatics derived from industrial activities often have functional groups such as alkyls, halogens, and nitro groups. Biodegradation is a major mechanism of removal of organic pollutants from a contaminated site. Syntrophorhabdus aromaticivorans is an obligate anaerobic bacterium that can grow on isophthalate and lives in syntrophy with sulfate-reducing Desulfovibrio species. When growing S. aromaticivorans with isophthalate, four proteins could be identified in high abundance by means of proteome analysis. One of these proteins, called isophthaloyl-CoA decarboxylase (IPCD), catalyzes the decarboxylation of isophthaloyl-CoA to benzoyl-CoA under anaerobic conditions.. In this study the encoding genes of IPCD from S. aromaticivorans were successfully cloned into different expression vectors to enable heterologous production in E. coli or Thauera chlorobenzoica.
The following results have been achieved:
1) Transformation of the gene sequences coding for the isophthaloyl-CoA decarboxylase (IPCD) enzyme with pRSF-Duet1 vector by restriction cloning or fast cloning into Escherichia coli NEB5α.
2) Transformation of the gene sequences coding for the isophthaloyl-CoA decarboxylase (IPCD) enzyme with pIZ1016 and POT1 into Thauera Chlorobenzoica.
3) Transferring and enhancing the genetic system of Thauera chlorobenzoica 3CB-1 strain adopting protocols of Thauera aromatica.
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