Is propionate alfa-oxidation involved in the reduction of biodegradable plastic production in Burkholderia sacchari? / Alfa-oxidação de propionato está envolvida na redução da produção de plástico biodegradável em Burkholderia sacchari?

AUTOR(ES)
DATA DE PUBLICAÇÃO

2008

RESUMO

Burkholderia sacchari is a new bacterial species from brazilian soil, able to grow in sucrose, accumulating intracellular granules of polyester belonging to the polyhydroxyalkanoate family (PHA). When cultivated on sucrose, the homopolymer poly-3-hydroxybutyrate is accumulated by this bacterium, which is used as biodegradable and biocompatible plastic. When sucrose and propionic acid are supplied as carbon sources, B. sacchari cells accumulate the copolymer poly-3-hydroxybutyrate-co-3-hydroxyvalerate (P3HB-co-3HV). However, a small percentage ofthe propionic acid supplied is converted to 3HV units, because efficient catabolic pathways convert this substrate preferentially to biomass, CO2 and water, thus reducing the efficiency of polymer production. At least two propionate catabolic pathways have been previously indicated in B. sacchari: a-oxidation and the 2-methylcitric acid (2MCC), the latter confirmed at molecular leveI. UV mutants previously obtained were unable to grow in propionate (prp) and also showed the phenotype affected concerning grow on intermediates of propionate a-oxidation. In the present work, after a screening in B. sacchari genomic libraries, one ofthem constructed also in the present work, the prp + phenotype was restored to the mutants by three different DNA fragments harbored by dones A), PI and P2. Quantitative experiments revealed that AI restored only partially the quantitative conversion of propionate to 3HV units to the mutants. PI restored the ability to grow in propionate and in other intermediates of a-oxidation to one prp mutant. A DNA 1.2 Kb subfragment of PI, still able to complement prp mutants, was subcloned and sequenced, showing similarity to DNA sequences encoding to LysR-type transcriptional regulators of various bacteria, including BlIrkholderia species. Adjacent regions to LysR in different genomes of BlIrkholderia are annotated as encoding to acyl-CoA dehydrogenases, neighboring a predicted acyl-CoA transferases/carnitine dehydratase and a permease ofthe major facilitator superfamily MFS-1. After confirmation ofthe same adjacent regions in B. sacchari and also their especific deletion, it will be possible to prove the presence of the pathway indicated here in the catabolism of propionate.

ASSUNTO(S)

burkholderia sacchari polihidroxialcanoatos applied microbiology poliester biodegradable plastic microbiologia aplicada plásticos biodegradáveis polyhydroxyalcanoates propionato polyester propionate burkholderia sacchari

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