Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Industrial production of semi-synthetic cephalosporins by Penicillium chrysogenum requires supplementation of the growth media with the side-chain precursor adipic acid. In glucose-limited chemostat cultures of P. chrysogenum, up to 88% of the consumed adipic acid was not recovered in cephalosporinrelated products, but used as an additional carbon and energy source for growth. This low efficiency of side-chain precursor incorporation provides an economic incentive for studying and engineering the metabolism of adipic acid in P. cluysogenum. Chemostat-based transcriptome analysis in the presence and absence of adipic acid confirmed that adipic acid metabolism in this fungus occurs via beta-oxidation. A set of 52 adipate-responsive genes included six putative genes for acyl-CoA oxidases and dehydrogenases, enzymes responsible for the first step of beta-oxidation. Subcellular localization of the differentially expressed acyl-CoA oxidases and dehydrogenases revealed that the oxidases were exclusively targeted to peroxisomes, while the dehydrogenases were found either in peroxisomes or in mitochondria. Deletion of the genes encoding the peroxisomal acyl-CoA oxidase Pc20g01800 and the mitochondrial acyl-CoA dehydrogenase Pc20g07920 resulted in a 1.6- and 3.7-fold increase in the production of the semi-synthetic cephalosporin intermediate adipoyl-6-APA, respectively. The deletion strains also showed reduced adipate consumption compared to the reference strain, indicating that engineering of the first step of beta-oxidation successfully redirected a larger fraction of adipic acid towards cephalosporin biosynthesis. (C) 2012 Elsevier Inc. All rights reserved.

Frutanos do tipo inulina e aumento da absorção de cálcio: uma revisão sistemática

Realizou-se uma revisão sistemática da literatura sobre os efeitos do consumo de frutanos do tipo inulina na absorção de cálcio. Resultados de quatro dos sete ensaios clínicos duplo-cegos controlados aleatorizados relatados neste trabalho indicam que o consumo diário da mistura de frutanos (inulina e oligofrutose 1:1) aumenta a absorção de cálcio, dependendo da idade cronológica e fisiológica, bem como do estado menopausal de indivíduos saudáveis que consomem quantidades adequadas de cálcio. O número limitado de ensaios clínicos realizados e as diferenças relativas ao planejamento experimental, tempo de estudo, tipo e quantidade de frutano consumido (inulina, oligofrutose ou a mistura de ambos), bem como a faixa etária dos indivíduos, impedem a generalização dos resultados observados. Esta revisão demonstra a necessidade de mais ensaios clínicos de longa duração, nos quais tanto a absorção de cálcio como a densidade mineral óssea sejam avaliadas. Futuros estudos devem contribuir para a compreensão dos mecanismos de ação dos frutanos no aumento da absorção de cálcio; para avaliar se esse efeito persiste em longo prazo e se pode ser considerado como benefício real para a saúde óssea; e para testar se tais efeitos poderiam beneficiar indivíduos de outras faixas etárias e diferentes condições fisiológicas. Evidências científicas consistentes e acumuladas ainda são necessárias para poder considerar o consumo de frutanos como uma estratégia de prevenção da osteoporose.

Partitioning and extraction of collagenase from Penicillium aurantiogriseum in poly(ethylene glycol)/phosphate aqueous two-phase system

Purification of collagenase produced by Penicillium aurantiogriseum URM4622 was carried using a PEG/phosphate aqueous two-phase system (ATPS). A 2(3)-full experimental design was used to investigate the influence of PEG molar mass, PEG concentration and phosphate concentration on the selected responses, namely partition coefficient, activity yield and purification factor. The ATPS was composed of PEG (molar mass of 550, 1500 and 4000 g/mol) at concentrations of 15.0, 17.5 and 20.0% (w/w) and phosphate at concentrations of 12.5, 15.0 and 17.5% (w/w). The best results of one-step extraction of collagenase from the fermentation broth (partition coefficient of 1.01, activity yield of 242% and purification factor of 23.5) were obtained at pH 6.0 using 20.0% (w/w) PEG 550 and 17.5% (w/w) phosphate. The results of this preliminary study demonstrate that the selected ATPS is satisfactorily selective for the extraction of such a collagenase. (C) 2012 Elsevier B.V. All rights reserved.