Bioprospecting and characterization of poly-bhydroxyalkanoate (PHAs) producing bacteria isolated from Colombian sugarcane producing areas

Abstract

Bioprospecting for poly-b-hydroxyalkanoate (PHA)-accumulating micro-organisms was carried out in sugarcane growing areas of Colombia. They were isolated in unbalanced culture medium (high
carbon/nitrogen ratio) with sucrose, fructose and glucose used as carbon source. PHAs producing bacteria were identified by staining with Sudan black and solubilising cellular components in sodium
hypochlorite. A arbitrary scale was established (ranging from 1 to 4) for selecting the best strains, acording to growth, staining with Sudan black and solubilising cellular material. 108 isolates rated higher than 2 were obtained by using this scale; 44 of these were selected for evaluating them in a balloon flask for their ability to grow in sucrose. Scheffe’s test grouped the isolates evaluated in the balloon flask for polymer production and productivity. The 6 best isolates were evaluated in a fermenter to determine their kinetic growth profiles, substrate consumption and polymer accumulation. Differential scanning calorimeter (DSC) was used on the recovered polymer for determining fusion temperature and the conclusion was reached that 2 strains accumulated poly--hydroxybutyrate (PHB) and another 4 accumulated hydroxy-butyrate copolymers and other monomer units. These 6 strains were molecularly characterised by partially sequencing the 16s rRNA ribosomal gene, localizing them in 4 clusters on the taxonomic tree.