Effect of pyruvate dehydrogenase complex deficiency on <Emphasis Type="SmallCaps">l</Emphasis>-lysine production with <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis>

Intracellular precursor supply is a critical factor for amino acid productivity of Corynebacterium glutamicum. To test for the effect of improved pyruvate availability on l-lysine production, we deleted the aceE gene encoding the E1p enzyme of the pyruvate dehydrogenase complex (PDHC) in the l-lysine-producer C. glutamicum DM1729 and characterised the resulting strain DM1729-BB1 for growth and l-lysine production. Compared to the host strain, C. glutamicum DM1729-BB1 showed no PDHC activity, was acetate auxotrophic and, after complete consumption of the available carbon sources glucose and acetate, showed a more than 50% lower substrate-specific biomass yield (0.14 vs 0.33 mol C/mol C), an about fourfold higher biomass-specific l-lysine yield (5.27 vs 1.23 mmol/g cell dry weight) and a more than 40% higher substrate-specific l-lysine yield (0.13 vs 0.09 mol C/mol C). Overexpression of the pyruvate carboxylase or diaminopimelate dehydrogenase genes in C. glutamicum DM1729-BB1 resulted in a further increase in the biomass-specific l-lysine yield by 6 and 56%, respectively. In addition to l-lysine, significant amounts of pyruvate, l-alanine and l-valine were produced by C. glutamicum DM1729-BB1 and its derivatives, suggesting a surplus of precursor availability and a further potential to improve l-lysine production by engineering the l-lysine biosynthetic pathway. This study is dedicated to Prof. Dr. Hermann Sahm on the occasion of his 65th birthday.     

Moraxella catarrhalis is internalized in respiratory epithelial cells by a trigger-like mechanism and initiates a TLR2- and partly NOD1-dependent inflammatory immune response

Moraxella catarrhalis is an important pathogen in patients with chronic obstructive lung disease (COPD). While M. catarrhalis has been categorized as an extracellular bacterium so far, the potential to invade human respiratory epithelium has not yet been explored. Our results obtained by electron and confocal microscopy demonstrated a considerable potential of M. catarrhalis to invade bronchial epithelial (BEAS-2B) cells, type II pneumocytes (A549) and primary small airway epithelial cells (SAEC). Moraxella invasion was dependent on cellular microfilament as well as on bacterial viability, and characterized by macropinocytosis leading to the formation of lamellipodia and engulfment of the invading organism into macropinosomes, thus indicating a trigger-like uptake mechanism. In addition, the cells examined expressed TLR2 as well as NOD1, a recently found cytosolic protein implicated in the intracellular recognition of bacterial cell wall components. Importantly, inhibition of TLR2 or NOD1 expression by RNAi significantly reduced the M. catarrhalis-induced IL-8 secretion. The role of TLR2 and NOD1 was further confirmed by overexpression assays in HEK293 cells. Overall, M. catarrhalis may employ lung epithelial cell invasion to colonize and to infect the respiratory tract, nonetheless, the bacteria are recognized by cell surface TLR2 and the intracellular surveillance molecule NOD1.     

A new and short convergent synthetic strategy to carbocyclic nucleosides

An efficient synthesis for racemic cyclopent-3-en-1-yl nucleoside analogues has been developed starting from cyclopentadiene. The key step is the regioselective hydroboration of a mixture of intermediate alkylatede cyclopentadienes to give one cyclopentenol.     

Physiological Responses of Contrasting Maize (Zea mays L.) Hybrids to Repeated Drought

Maize is the most important crop worldwide in terms of production and yield, but every year a considerable amount of yield is lost due to drought. The foreseen increase in the number of drought spells due to climate change raises the question whether the ability to recover quickly after a water pulse may be a relevant trait for overall drought resistance. We here address the following hypotheses: (i) different maize hybrids exhibit distinct physiological adaptive responses to drought stress and (ii) these responses affect the ability to recover from the stress. (iii) The relative biomass production of maize hybrids, which show severe drought symptoms but are able to recover quickly after a water pulse, is comparable to those hybrids, which invest more energy into tolerance mechanisms. The physiological responses of eight maize hybrids to repeated drought were elucidated employing physiological parameters such as electrolyte leakage, osmolality, relative water content, growth rate and gas-exchange measurements. Only one hybrid was able to maintain biomass production under drought conditions. Amongst the others, two hybrids with similar growth inhibition but contrasting physiological responses were identified by a PCA analysis. Both strategies, i.e. stabilization of leaf water content via resistance mechanisms versus high recovery potential were equally effective in maintaining aboveground biomass production in the scenario of a long drought intermitted by a water-pulse. However, each strategy might be advantageous under different drought stress scenarios. Overall, the recovery potential is underestimated in drought resistance under natural conditions, which includes periodic cycles of drought and rewatering, and should be considered in screening trials.

     

Oil and gas development in the World Heritage and wider protected area network in sub-Saharan Africa

More than 25% of natural World Heritage (WH) sites worldwide are estimated to be under pressure from existing or future mining and energy activities (IUCN 2008; UNESCO 2009). However, that ‘pressure’ has yet to be quantitatively defined and assessed for many regions of the world. We conducted a GIS-based analysis of overlap and proximity between natural WH sites and areas allocated to oil and gas concessions as well as pipelines and oil wells for all of sub-Saharan Africa. We found that oil and gas concessions were located within 27% of the WH sites, though no currently active oil wells were operating directly within the WH sites. A proximity-based assessment of oil and gas concessions within 5 km of WH site boundaries included only one additional WH site, suggesting that sites susceptible to indirect impacts from oil and gas development are likely to be those already overlapped by concessions. Our findings indicate that activity from oil and gas development in sub-Saharan WH sites has to date been limited; however, future pressure cannot be ruled out, due to continued presence of concessions within more than one quarter of the network, and projected expansion of oil and gas exploration within the region. Our results may be used to inform the inclusion of new sites into the WH network.     

BRAF mutations in cutaneous melanoma are independently associated with age, anatomic site of the primary tumor, and the degree of solar elastosis at the primary tumor site

Oncogenic BRAF mutations are more frequent in cutaneous melanoma occurring at sites with little or moderate sun-induced damage than at sites with severe cumulative solar ultraviolet (UV) damage. We studied cutaneous melanomas from geographic regions with different levels of ambient UV radiation to delineate the relative effects of cumulative UV damage, age, and anatomic site on the frequency of BRAF mutations. We show that BRAF-mutated melanomas occur in a younger age group on skin without marked solar elastosis and less frequently affect the head and neck area, compared to melanomas without BRAF mutations. The findings indicate that BRAF-mutated melanomas arise early in life at low cumulative UV doses, whereas melanomas without BRAF mutations require accumulation of high UV doses over time. The effect of anatomic site on the mutation spectrum further suggests regional differences among cutaneous melanocytes.