Promoting microbial utilization of phenolic substrates from bio-oil

Journal of Industrial Microbiology & Biotechnology - The economic viability of the biorefinery concept is limited by the valorization of lignin. One possible method of lignin valorization is...     

Risk of otitis externa after swimming in recreational fresh water lakes containing Pseudomonas aeruginosa.

OBJECTIVE--To determine whether an outbreak of otitis externa was due to bathing in recreational fresh water lakes and to establish whether the outbreak was caused by Pseudomonas aeruginosa in the water. DESIGN--Matched case-control study. SETTING--The Achterhoek area, the Netherlands. SUBJECTS--98 cases with otitis externa and 149 controls matched for age, sex, and place of residence. MAIN OUTCOME MEASURES--Odds ratios for type of swimming water and frequency of swimming; presence of P aeruginosa in ear swabs and fresh water lakes. RESULTS--Otitis externa was strongly associated with swimming in recreational fresh water lakes in the previous two weeks (odds ratio 15.5 (95% confidence interval) 4.9 to 49.2) compared with non-swimming). The risk increased with the number of days of swimming, and subjects with recurrent ear disease had a greatly increased risk. The lakes met the Dutch bathing water standards and those set by the European Commission for faecal pollution in the summer of 1994, but P aeruginosa was isolated from all of them, as well as from the ear swabs of 78 (83%) of the cases and 3 (4%) of the controls. CONCLUSIONS--Even when current bathing water standards are met, swimming can be associated with a substantial risk of otitis externa because of exposure to P aeruginosa. People with recurrent ear disease should take special care when swimming in waters containing P aeruginosa.     

Crop Management Impacts Biofuel Quality: Influence of Switchgrass Harvest Time on Yield, Nitrogen and Ash of Fast Pyrolysis Products

Although upgrading bio-oil from fast pyrolysis of biomass is an attractive pathway for biofuel production, nitrogen (N) and mineral matter carried over from the feedstock to the bio-oil represents a serious contaminant in the process. Reducing the N and ash content of biomass feedstocks would improve process reliability and reduce production costs of pyrolytic biofuels. This study investigated: (1) How does switchgrass harvest date influence the yield, N concentration ([N]), and ash concentration of biomass and fast pyrolysis products? and (2) Is there a predictive relationship between [N] of switchgrass biomass and [N] of fast pyrolysis products? Switchgrass from five harvest dates and varying [N] from central Iowa were pyrolyzed using a free-fall reactor. Harvestable biomass peaked in August (8.6 Mg ha^?1), dropping significantly by November (6.7 Mg ha^?1, P?=?0.0027). Production of bio-oil per unit area mirrored that of harvested biomass at each harvest date; however, bio-oil yield per unit dry biomass increased from 46.6 % to 56.7 % during the season (P?=?0.0018). Allowing switchgrass to senesce lowered biomass [N] dramatically, by as much as 68 % from June to November (P?<?0.0001). Concurrently, bio-oil [N] declined from 0.51 % in June to 0.17 % by November (P?<?0.0001). Significant reductions in ash concentration were also observed in biomass and char. Finally, we show for the first time that the [N] of switchgrass biomass is a strong predictor of the [N] of bio-oil, char, and non-condensable gas with R ² values of 0.89, 0.94, and 0.88, respectively.     

Production of biorenewable styrene: utilization of biomass-derived sugars and insights into toxicity

Fermentative production of styrene from glucose has been previously demonstrated in Escherichia coli. Here, we demonstrate the production of styrene from the sugars derived from lignocellulosic biomass depolymerized by fast pyrolysis. A previously engineered styrene-producing strain was further engineered for utilization of the anhydrosugar levoglucosan via expression of levoglucosan kinase. The resulting strain produced 240 ± 3 mg L^?1 styrene from pure levoglucosan, similar to the 251 ± 3 mg L^?1 produced from glucose. When provided at a concentration of 5 g L^?1, pyrolytic sugars supported styrene production at titers similar to those from pure sugars, demonstrating the feasibility of producing this important industrial chemical from biomass-derived sugars. However, the toxicity of contaminant compounds in the biomass-derived sugars and styrene itself limit further gains in production. Styrene toxicity is generally believed to be due to membrane damage. Contrary to this prevailing wisdom, our quantitative assessment during challenge with up to 200 mg L^?1 of exogenously provided styrene showed little change in membrane integrity; membrane disruption was observed only during styrene production. Membrane fluidity was also quantified during styrene production, but no changes were observed relative to the non-producing control strain. This observation that styrene production is much more damaging to the membrane integrity than challenge with exogenously supplied styrene provides insight into the mechanism of styrene toxicity and emphasizes the importance of verifying proposed toxicity mechanisms during production instead of relying upon results obtained during exogenous challenge.     

Callus ontogeny of the Kappaphycus alvarezii (Rhodophyta, Gigartinales) brown tetrasporophyte strain

This study analyzes the callus ontogeny of the Kappaphycus alvarezii brown strain tetrasporophyte using micropropagation and tissue culture. Calli were induced from axenic explants and grown for 2 months. Ontogenic analysis was made in explants collected from days?7 to 60 of culture and observed under light and transmission electron microscopy (TEM). At the end of the experimental period, 60-day samples were also analyzed by confocal microscopy. Formation of calli was initiated in the first week from medullary and cortical cells of the explant region in air contact. Alterations in the apical region of the cells were observed in TEM as cell wall thickening, proliferation of convoluted membranes, increased number of mitochondria and chloroplast alterations, indicating the dedifferentiation of these cells. Throughout the experiment, the thick cell wall of the cells which compose callus filaments showed the presence of acidic polysaccharides, suggesting a large amount of carrageenan and neutral polysaccharides. The cytoplasm showed the following main characteristics: large amount of starch grains, presence of chloroplasts with altered plastoglobules, as well as disorganized thylakoids, presence of several convoluted membranes, and vacuoles. These filaments maintained their uniseriate organization and irregular branching with many intercellular connections between them. Despite the initial stress caused by explant isolation and the beginning of callus formation, results showed that filaments were capable of reorganization and development, retaining their dedifferentiated cells, which, in turn, strongly suggested the potential of callus-based micropropagation.     

Damage to the microbial cell membrane during pyrolytic sugar utilization and strategies for increasing resistance

Lignocellulosic biomass is an appealing feedstock for the production of biorenewable fuels and chemicals, and thermochemical processing is a promising method for depolymerizing it into sugars. However, trace compounds in this pyrolytic sugar syrup are inhibitory to microbial biocatalysts. This study demonstrates that hydrophobic inhibitors damage the cell membrane of ethanologenic Escherichia coli KO11+lgk. Adaptive evolution was employed to identify design strategies for improving pyrolytic sugar tolerance and utilization. Characterization of the resulting evolved strain indicates that increased resistance to the membrane-damaging effects of the pyrolytic sugars can be attributed to a glutamine to leucine mutation at position 29 of carbon storage regulator CsrA. This single amino acid change is sufficient for decreasing EPS protein production and increasing membrane integrity when exposed to pyrolytic sugars.     

Pronounced chemical response of Subarctic lakes to climate‐driven losses in surface area

Losses in lake area have been observed for several Arctic and Subarctic regions in recent decades, with unknown consequences for lake ecosystems. These reductions are primarily attributed to two climate‐sensitive mechanisms, both of which may also cause changes in water chemistry: (i) increased imbalance of evaporation relative to inflow, whereby increased evaporation and decreased inflow act to concentrate solutes into smaller volumes; and (ii) accelerated permafrost degradation, which enhances sublacustrine drainage while simultaneously leaching previously frozen solutes into lakes. We documented changes in nutrients [total nitrogen (TN), total phosphorus (TP)] and ions (calcium, chloride, magnesium, sodium) over a 25 year interval in shrinking, stable, and expanding Subarctic lakes of the Yukon Flats, Alaska. Concentrations of all six solutes increased in shrinking lakes from 1985–1989 to 2010–2012, while simultaneously undergoing little change in stable or expanding lakes. This created a present‐day pattern, much weaker or absent in the 1980s, in which shrinking lakes had higher solute concentrations than their stable or expanding counterparts. An imbalanced evaporation‐to‐inflow ratio (E/I) was the most likely mechanism behind such changes; all four ions, which behave semiconservatively and are prone to evapoconcentration, increased in shrinking lakes and, along with TN and TP, were positively related to isotopically derived E/I estimates. Moreover, the most conservative ion, chloride, increased >500% in shrinking lakes. Conversely, only TP concentration was related to probability of permafrost presence, being highest at intermediate probabilities. Overall, the substantial increases of nutrients (TN >200%, TP >100%) and ions (>100%) may shift shrinking lakes towards overly eutrophic or saline states, with potentially severe consequences for ecosystems of northern lakes.