Kinetics of the aerobic co-metabolism of 1,1-dichloroethylene by Achromobacter sp.: a novel benzene-grown culture

Batch experiments were performed for the aerobic co-metabolism of 1,1-dichloroethylene (1,1-DCE) by Achromobacter sp., identified by gene sequencing of 16S rRNA and grown on benzene. Kinetic models were employed to simulate the co-metabolic degradation of 1,1-DCE, and relevant parameters were obtained by non-linear least squares regression. Benzene at 90 mg L^?1 non-competitively inhibited degradation of 1,1-DCE (from 125 to 1,200 μg L^?1). The maximum specific utilization (k_c) rate and the half-saturation constant (K_c) for 1,1-DCE were 54 ± 0.85 μg h^?1 and 220 ± 6.8 μg L^?1, respectively; the k_b and K_b for benzene were 13 ± 0.18 mg h^?1 and 28 ± 0.42 mg L^?1, respectively. This study provides a theoretical basis to predict the natural attenuation when benzene and 1,1-DCE occur as co-contaminants.     

Enhancing ethanol production from cellulosic sugars using <Emphasis Type="Italic">Scheffersomyces (Pichia) stipitis</Emphasis>

Studies were performed on the effect of CaCO₃ and CaCl₂ supplementation to fermentation medium for ethanol production from xylose, glucose, or their mixtures using Scheffersomyces (Pichia) stipitis. Both of these chemicals were found to improve maximum ethanol concentration and ethanol productivity. Use of xylose alone resulted in the production of 20.68 ± 0.44 g L^?1 ethanol with a productivity of 0.17 ± 0.00 g L^?1 h^?1, while xylose plus 3 g L^?1 CaCO₃ resulted in the production of 24.68 ± 0.75 g L^?1 ethanol with a productivity of 0.21 ± 0.01 g L^?1 h^?1. Use of xylose plus glucose in combination with 3 g L^?1 CaCO₃ resulted in the production of 47.37 ± 0.55 g L^?1 ethanol (aerobic culture), thus resulting in an ethanol productivity of 0.39 ± 0.00 g L^?1 h^?1. These values are 229 % of that achieved in xylose medium. Supplementation of xylose and glucose medium with 0.40 g L^?1 CaCl₂ resulted in the production of 44.84 ± 0.28 g L^?1 ethanol with a productivity of 0.37 ± 0.02 g L^?1 h^?1. Use of glucose plus 3 g L^?1 CaCO₃ resulted in the production of 57.39 ± 1.41 g L^?1 ethanol under micro-aerophilic conditions. These results indicate that supplementation of cellulosic sugars in the fermentation medium with CaCO₃ and CaCl₂ would improve economics of ethanol production from agricultural residues.     

Concentration,Distribution, Source,and Risk Assessment of PAHs and Heavy Metals in Surface Water from the Three Gorges Reservoir,China

Fifteen polycyclic aromatic hydrocarbons (PAHs) and heavy metals (Cr, Ni, As, Cd, Pb, and Hg) were quantified in 19 surface water sites of the Three Gorges Reservoir, China. The total concentrations of 15 PAHs and six heavy metals in the 19 sample sites ranged from 130.8 ng L^?1 to 227.5 ng L^?1 and 3.2 μg L^?1 to 6.0 μg L^?1, respectively. The mean concentration of As was the highest among the six heavy metals (2.1 ± 0.3 μg L^?1), followed by Cr (0.5 ± 0.3 μg L^?1), Ni (1.3 ± 0.1 μg L^?1), Cd (0.2 ± 0.01 μg L^?1), Pb (0.07 ± 0.08 μ g L^?1) and Hg (0.05 ± 0.08 μg L^?1). The isomer ratio results suggest that PAHs at most sites were mainly from petroleum combustion, while coal and biomass combustion was the main source at sites 1, 2, 6, 7, 9, 14, and 17. Based on principal component analysis, the main source of heavy metals was anthropogenic activities and weathering of bedrocks. Depending on characteristic of RQ_(NCs) ≥ 1 and RQ_(MPCs) < 1, BaA showed higher potential ecological risk than other PAHs, therefore, all sampling site needed to be paid much more attention, included some remedial actions. Meanwhile, after assessing human health risk of heavy metal, it was unlikely to experience adverse health effects, even exposing through more pathways and six kinds of heavy metals simultaneously.     

Copper and thermal perturbations on the early life processes of the hard coral <Emphasis Type="Italic">Platygyra acuta</Emphasis>

Anthropogenic pollutants and climate change are major threats to coral reefs today. Yet interactions between chemical and thermal perturbations have not been fully explored in reef studies. Here, we present the single and combined effects of copper (Cu) with thermal stress on five early life-history stages/processes (fertilization, larval mortality, swimming ability, metamorphosis and growth of juvenile recruits) of the massive coral Platygyra acuta in Hong Kong. In the first four experiments, coral gametes and larvae were exposed to different Cu doses (0–200 μg L^?1, apart from the fertilization assay in which 0–1000 μg L^?1 was used) and temperature treatments (ambient and ambient +2 or +3 °C as a thermal stress treatment) following a factorial experimental design. Exposure time was 5 h for the fertilization assay and 48 h for the other experiments. The last experiment on growth of coral recruits was conducted over 56 d with 0–80 μg L^?1 Cu used. Cu significantly reduced percent fertilization success, percentage of active swimming larvae and larval survivorship (EC₅₀s, the half maximal effective concentrations, for percent fertilization success and percentage of active swimming larvae were 92–145 and 45–47 μg L^?1 respectively. While LC₅₀, the lethal concentration that kills 50% of the population, was 101–110 μg L^?1), while growth of coral recruits was not affected at 80 μg L^?1 Cu for 56 d. No settling cues were used in the settlement experiment. In their absence, percent metamorphosis increased with Cu doses, in sharp contrast to earlier findings. Settlement and metamorphosis may thus be strategies for coral larvae to escape from Cu toxicity. Thermal treatment did not significantly affect any experimental end points. This is likely because the thermal regimes used in the experiments were within the range experienced by local corals. The high variability in Cu toxicities indicates differential susceptibilities of the various life-history stages/processes of P. acuta. The level of Cu tolerance was also markedly higher than that reported in the literature for other coral species. This provides evidence to suggest possible adaptation of this species to survive in a highly polluted marine environment like that in Hong Kong.     

Physiological responses and accumulation of pollutants in woody species under in situ polluted condition in Southern China

Physiological leaf traits and accumulation of pollutants of ten woody species in response to air pollution at seriously polluted site Sanguigang (SGG) and control site Maofengshan (MFS) in Southern China were studied. Net photosynthetic rates of most species at SGG were lower than those at MFS, but stomatal conductance (g_s) showed opposite trend. The specific leaf area of Aporusa dioica, Sapium discolor, Schefflera octophylla and Toxicodendron succedaneum were significantly, 46.77, 13.09, 55.11 and 23.51 %, higher in SGG than in MFS, while chlorophyll content being the opposite. A. dioica had the highest sulphur (S) content at both sites (11.74 mg g^?1 at SGG and 11.07 mg g^?1 at MFS). Heavy metals concentrations were generally higher in species at SGG than at MFS. S. octophylla showed significantly higher concentrations of Zn, Cd and Mn (341.81, 2.41 and 2,287.29 μg g^?1) than other species at SGG. Moreover, A. dioica had the highest Pb concentration (9.19 μg g^?1), and L. glutinosa showed the highest Cr concentration (3.40 μg g^?1). According to the integrated results, we infer that A. dioica, S. octophylla and L. glutinosa are the promising species for phytoremediation in the ceramic industry polluted environment.     

Improvement of 5-aminolevulinic acid production by Rubrivivax benzoatilyticus PS-5 with self-flocculation by co-fermentation of precursors and volatile fatty acids under pH-controlled conditions

Photosynthetic bacteria are known to utilize volatile fatty acids as a carbon source for growth and product formation. In this study, a new isolate, Rubrivivax benzoatilyticus PS-5, possessing self-flocculation properties, was cultivated in modified glutamate-malate (GM) medium containing glutamate and malate as carbon sources. The effect of acetic acid, propionic acid and butyric acid (at 1–4 g L^?1) as co-substrates and 7.5 mM glycine, 10 mM succinic acid as precursors for 5-aminolevulinic acid (ALA) production from R. benzoatilyticus PS-5 was investigated. Among the volatile fatty acids tested, acetic acid was preferred to butyric acid and propionic acid, with the optimum concentrations of 3 g L^?1, 1 g L^?1 and 3 g L^?1, respectively. The highest ALA production was 169.71 μM, 162.16 μM and 46.18 μM, respectively, while the highest productivity was 2.57 μM h^?1, 2.25 μM h^?1 and 0.96 μM h^?1, respectively. The precursor was consumed completely (100 %) while the assimilation of the acetic acid and butyric acid was 62.50 % and 48.65 %, respectively. Supplementation of propionic acid (at 1–4 g l^?1) had a negative effect on growth and ALA production. To increase production efficiency, the pH-control strategy (at pH 6.0–8.0) during fermentation was tested. The optimum pH was 7.0, giving the maximum ALA production of 286.18 μM and a productivity of 3.97 μM h^?1. These values were 1.68-fold and 1.54-fold higher, respectively, than those under uncontrolled pH conditions.     

Antiviral Action of Methylated β-Lactoglobulin on the Human Influenza Virus A Subtype H3N2

Antiviral activity of methylated β-lactoglobulin (Met-BLG) against H3N2 infected into MDCK cell lines depended on concentration of Met-BLG, viral load, and duration of infection. IC_50% of the hemagglutination activity for 1 and 0.2 MOI (multiplicity of infection) after 24 h of incubation at 37 °C in the presence of 5% CO₂ were 20 ± 0.8 and 17 ± 0.7 μg mL^?1 Met-BLG, respectively. Longer incubation period (4 days) was associated with low IC_50% of the hemagglutination activity (7.1 ± 0.3 μg mL^?1 Met-BLG) and low IC_50% of immuno-fluorescence of viral nucleoproteins (9.7 ± 0.4 μg mL^?1 Met-BLG) when using 0.2 and 0.1 MOI, respectively. A concentration of 25 μg mL^?1 of Met-BLG reduced the amount of replicating virus by about 2 and 1.3 logs when the viral load was 0.01 and 0.1 MOI, respectively, while higher concentrations reduced it by about 5–6 logs. Antiviral action of Met-BLG was coupled with a cellular protective action, which reached 100% when using 0.01 and 0.1 MOI and 83% when using 1.0 MOI. The time of Met-BLG addition after the viral infection was determinant for its antiviral efficacy and for its protection of the infected MDCK cell lines. Anti-hemagglutination action and cell protective action decreased gradually and in parallel with the delay in the time of Met-BLG addition to disappear totally after 10 h delay.     

Effects of manganese on the microstructures of Chenopodium ambrosioides L., A manganese tolerant plant

Chenopodium ambrosioides L. can tolerate high concentrations of manganese and has potential for its use in the revegetation of manganese mine tailings. Following a hydroponic investigation, transmission electron microscopy (TEM)-energy disperse spectroscopy (EDS) was used to study microstructure changes and the possible accumulation of Mn in leaf cells of C. ambrosioides in different Mn treatments (200, 1000, 10000 μmol·L^?1). At 200 μmol·L^?1, the ultrastructure of C. ambrosioides was clearly visible without any obvious damage. At 1000 μmol·L^?1, the root, stem and leaf cells remained intact, and the organelles were clearly visible without any obvious damage. However, when the Mn concentration exceeded 1000 μmol·L^?1 the number of mitochondria in root cells decreased and the chloroplasts in stem cells showed a decrease in grana lamellae and osmiophilic granules. Compared to controls, treatment with 1000 μmol·L^?1 or 10000 μmol·L^?1 Mn over 30 days, gave rise to black agglomerations in the cells. At 10000 μmol·L^?1, Mn was observed to form acicular structures in leaf cells and intercellular spaces, which may be a form of tolerance and accumulation of Mn in C. ambrosioides. This study has furthered the understanding of Mn tolerance mechanisms in plants, and is potential for the revegetation of Mn-polluted soils.     

<Emphasis Type="Italic">In vitro</Emphasis> sucrose concentration influences microtuber production and diosgenin content in white yam (<Emphasis Type="Italic">Dioscorea rotundata</Emphasis> Poir)

Dioscorea spp. is an important food crop in many countries and the source of the phytochemical diosgenin. Efficient microtuber production could provide source materials for farm-planting stock, for food markets, and for the production of high-diosgenin-producing cultivars. The first step in this study was optimizing the plant growth regulators for plantlet production, followed by a study of the effects of sucrose concentration on microtuber induction and diosgenin production. Significantly, more shoots (3.5) were produced at 4.65 μM (1 mg L^?1) kinetin (KIN), longer shoots (4.1 cm) were obtained at 2.46 μM (0.5 mg L^?1) indole-3-butyric acid (IBA), and root number (3.9) was significantly higher at 5.38 μM (1 mg L^?1) naphthalene acetic acid (NAA) than in other treatments. Increased sucrose concentrations in the optimized growth medium with 4.65 μM KIN and 5.38 μM NAA had significant effects on microtuber production (p < 0.01) and diosgenin content (p < 0.05). The most microtubers (6.2) were obtained with 100 g L^?1 sucrose, while those on 80 g L^?1 sucrose were the heaviest (0.7 g) and longest (7.4 mm). Microtubers formed in medium with 80 g L^?1 sucrose had significantly higher diosgenin content (3.64% [w/w]) than those in other sucrose treatments (< 2%) and was similar to that of field-grown parent tubers (3.79%). This result indicates an important role for sucrose in both microtuber growth and diosgenin production. Medium containing 4.65 μM KIN and 5.38 μM NAA is recommended for plantlet production, and medium containing 80 g L^?1 sucrose is recommended for microtuber and diosgenin production.     

Circadian rhythms of melatonin in haemolymph and optic lobes of Chinese mitten crab (Eriocheir sinensis) and Chinese grass shrimp (Palaemonetes sinensis)

This study is the first to examine the circadian rhythms of melatonin in Eriocheir sinensis and Palaemonetes sinensis, two economically important crustaceans. We collected haemolymph and optic lobes from both species every 4 h over a whole day cycle. Melatonin content was measured with high-performance liquid chromatography. E. sinensis haemolymph exhibited significant (p < 0.05) peaks in melatonin at 16:00 (0.180 ± 0.020 μg·mL^?1) and 24:00 (0.244 ± 0.055 μg·mL^?1), while eyestalks had significant peaks at 16:00 (72.377 ± 18.100 μg·eyestalk^?1) and 24:00 (98.756 ± 30.271 μg·eyestalk^?1). In P. sinensis, melatonin peaked significantly only at 16:00 in optic lobes (12.493 ± 1.475 μg·eyestalk^?1) (p < 0.05); no significant peaks were present in haemolymph. Thus, both E. sinensis and P. sinensis exhibit species-specific melatonin rhythms. Time of day should therefore be considered when examining the physiological status of both crustaceans, given the potential influence of fluctuating daily melatonin concentrations.     

Ecosystem respiration derived from 222Rn measurements on Rishiri Island, Japan

We evaluated the nighttime CO₂ flux (ecosystem respiration) on Rishiri Island, located at the northern tip of Hokkaido, Japan, from 2009 to 2011, by using the relationship between atmospheric ²²²Rn and CO₂ concentrations. The annual mean CO₂ flux was 1.8 μmol m^?2 s^?1, with a maximum monthly mean in July (4.6 ± 2.6 μmol m^?2 s^?1) and a broad minimum from December to March (0.33 ± 0.29 μmol m^?2 s^?1). The annual mean was comparable to fluxes at the JapanFlux sites in northern Japan. During the season of snow cover (mid-December to early April), the CO₂ flux was low (0.45 ± 0.43 μmol m^?2 s^?1). Total annual respiration was estimated at 679 ± 174 g cm^?2, about 8 % of which occurred during the season of snow cover.     

Elicitor mediated enhancement of wedelolactone in cell suspension culture of <Emphasis Type="Italic">Eclipta alba</Emphasis> (L.) Hassk

The present research focused on enhancing the production of wedelolactone through cell suspension culture (CSC) in Eclipta alba (L.) Hassk. With an aim of attaining a sustainable CSC, various plant growth regulators, elicitors and agitation speed were examined. Nodal segments of in vitro propagated plantlets induced the maximum percentage (93.47?±?0.61%) of callus inoculated on Murashige and Skoog (MS) medium fortified with picloram (2 mg L^?1). The growth kinetics of CSC exhibited a sigmoid pattern with a lag phase (0–6 days), a log phase (6–18 days), a stationary phase (18–24 days) and then death phase thereafter. The highest biomass accumulation in CSC with 7.09?±?0.06 g 50 mL^?1 fresh weight, 1.52?±?0.02 g 50 mL^?1 dry cell weight, 1.34?±?0.01?×?10⁶ cell mL^?1 total cell count and 57.00?±?0.58% packed cell volume was obtained in the liquid MS medium supplemented with 1.5 mg L^?1 picloram plus 0.5 mg L^?1 kinetin at 120 rpm. High performance thin layer chromatography confirmed that yeast extract (biotic elicitor) at 150 mg L^?1 accumulated more CSC biomass with 1.22-fold increase in wedelolactone (288.97?±?1.94 µg g^?1 dry weight) content in comparison to the non-elicited CSC (237.78?±?0.04 µg g^?1 dry weight) after 120 h of incubation. Contrastingly, methyl jasmonate (abiotic elicitor) did not alter the biomass but increased the wedelolactone content (259.32?±?1.06 µg g^?1 dry weight) to an extent of 1.09-fold at 100 µM. Complete plantlet regeneration from CSC was possible on MS medium containing N⁶-benzyladenine (0.75 mg L^?1) and abscisic acid (0.5 mg L^?1). Thus, the establishment of protocol for CSC constitutes the bases for future biotechnological improvement studies in this crop.     

Gaseous and fluvial carbon export from an Amazon forest watershed

The transfer of carbon (C) from Amazon forests to aquatic ecosystems as CO₂ supersaturated in groundwater that outgases to the atmosphere after it reaches small streams has been postulated to be an important component of terrestrial ecosystem C budgets. We measured C losses as soil respiration and methane (CH₄) flux, direct CO₂ and CH₄ fluxes from the stream surface and fluvial export of dissolved inorganic C (DIC), dissolved organic C (DOC), and particulate C over an annual hydrologic cycle from a 1,319-ha forested Amazon perennial first-order headwater watershed at Tanguro Ranch in the southern Amazon state of Mato Grosso. Stream pCO₂ concentrations ranged from 6,491 to 14,976 ??atm and directly-measured stream CO₂ outgassing flux was 5,994 ± 677 g C m^?2 y^?1 of stream surface. Stream pCH₄ concentrations ranged from 291 to 438 ??atm and measured stream CH₄ outgassing flux was 987 ± 221 g C m^?2 y^?1. Despite high flux rates from the stream surface, the small area of stream itself (970 m², or 0.007% of watershed area) led to small directly-measured annual fluxes of CO₂ (0.44 ± 0.05 g C m² y^?1) and CH₄ (0.07 ± 0.02 g C m² y^?1) per unit watershed land area. Measured fluvial export of DIC (0.78 ± 0.04 g C m^?2 y^?1), DOC (0.16 ± 0.03 g C m^?2 y^?1) and coarse plus fine particulate C (0.001 ± 0.001 g C m^?2 y^?1) per unit watershed land area were also small. However, stream discharge accounted for only 12% of the modeled annual watershed water output because deep groundwater flows dominated total runoff from the watershed. When C in this bypassing groundwater was included, total watershed export was 10.83 g C m^?2 y^?1 as CO₂ outgassing, 11.29 g C m^?2 y^?1 as fluvial DIC and 0.64 g C m^?2 y^?1 as fluvial DOC. Outgassing fluxes were somewhat lower than the 40?C50 g C m^?2 y^?1 reported from other Amazon watersheds and may result in part from lower annual rainfall at Tanguro. Total stream-associated gaseous C losses were two orders of magnitude less than soil respiration (696 ± 147 g C m^?2 y^?1), but total losses of C transported by water comprised up to about 20% of the ± 150 g C m^?2 (±1.5 Mg C ha^?1) that is exchanged annually across Amazon tropical forest canopies.     

Manipulation of culture strategies to enhance capsaicin biosynthesis in suspension and immobilized cell cultures of Capsicum chinense Jacq. cv. Naga King Chili

Manipulation of culture strategies was adopted to study the influence of nutrient stress, pH stress and precursor feeding on the biosynthesis of capsaicin in suspension and immobilized cell cultures of C. chinense. Cells cultured in the absence of one of the four nutrients (ammonium and potassium nitrate for nitrate and potassium stress, potassium dihydrogen orthophosphate for phosphorus stress, and sucrose for sugar stress) influenced the accumulation of capsaicin. Among the stress factors studied, nitrate stress showed maximal capsaicin production on day 20 (505.9 ± 2.8 μg g^?1 f.wt) in immobilized cell, whereas in suspension cultures the maximum accumulation (345.5 ± 2.9 μg g^?1 f.wt) was obtained on day 10. Different pH affected capsaicin accumulation; enhanced accumulation of capsaicin (261.6 ± 3.4 μg g^?1 f.wt) was observed in suspension cultures at pH 6 on day 15, whereas in case of immobilized cultures the highest capsaicin content (433.3 ± 3.3 μg g^?1 f.wt) was obtained at pH 5 on day 10. Addition of capsaicin precursors and intermediates significantly enhanced the biosynthesis of capsaicin, incorporation of vanillin at 100 μM in both suspension and immobilized cell cultures resulted in maximum capsaicin content with 499.1 ± 5.5 μg g^?1 f.wt on day 20 and 1,315.3 ± 10 μg g^?1 f.wt on day 10, respectively. Among the different culture strategies adopted to enhance capsaicin biosynthesis in cell cultures of C. chinense, cells fed with vanillin resulted in the maximum capsaicin accumulation. The rate of capsaicin production was significantly higher in immobilized cells as compared to freely suspended cells.     

Process Engineering for High-Cell-Density Cultivation of Lipid Rich Microalgal Biomass of <Emphasis Type="Italic">Chlorella</Emphasis> sp. FC2 IITG

In the present study, process engineering strategy was applied to achieve lipid-rich biomass with high density of Chlorella sp. FC2 IITG under photoautotrophic condition. The strategy involved medium optimization, intermittent feeding of limiting nutrients, dynamic change in light intensity, and decoupling growth and lipid induction phases. Medium optimization was performed using combinations of artificial neural network or response surface methodology with genetic algorithm (ANN-GA and RSM-GA). Further, a fed-batch operation was employed to achieve high cell density with intermittent feeding of nitrate and phosphate along with stepwise increase in light intensity. Finally, mutually exclusive biomass and lipid production phases were decoupled into two-stage cultivation process: biomass generation in first stage under nutrient sufficient condition followed by lipid enrichment through nitrogen starvation. The key findings were as follows: (i) ANN-GA resulted in an increase in biomass titer of 157 % (0.95 g L^?1) in shake flask and 42.8 % (1.0 g L^?1) in bioreactor against unoptimized medium at light intensity of 20 μE m^?2 s^?1; (ii) further optimization of light intensity in bioreactor gave significantly improved biomass titer of 5.6 g L^?1 at light intensity of 250 μE m^?2 s^?1; (iii) high cell density of 13.5 g L^?1 with biomass productivity of 675 mg L^?1 day^?1 was achieved with dynamic increase in light intensity and intermittent feeding of limiting nutrients; (iv) finally, two-phase cultivation resulted in biomass titer of 17.7 g L^?1 and total lipid productivity of 313 mg L^?1 day^?1 which was highest among Chlorella sp. under photoautotrophic condition.     

Culture Media Statistical Optimization for Biomass Production of a Ligninolytic Fungus for Future Rice Straw Degradation

The main objective of this study was to optimize a culture media for low scale biomass production of Pleurotus spp. Future applications of this optimization will be implemented for “in situ” rice straw degradation, increase soil nutrients availability, and lower residue and rice culture management costs. Soil samples were taken from different points in six important rice production cities in Colombia. For carbon and nitrogen source selection a factorial 4² design was carried out. The Plackett-Burman design permitted to detect carbon, nitrogen and inducer effects on fungus growth (response variable for all designs). This optimization was carried out by a Box-Behnken design. Finally a re-optimization assay for glucose concentration was performed by means of a One Factor design. Only 4/33 (12 %) isolates showed and important laccase or manganese peroxidase activity compared to Pleurotus ostreatus (HPB/P3). We obtained an increased biomass production in Pleurotus spp. (T1.1.) with glucose, followed by rice husk. Rice straw was considered an inducing agent for lignin degradation. Glucose was a significant component with positive effects, whereas Tween 80 and pH had negative effects. On the contrary, rice husk, yeast extract and CaCl₂ were not significant components for increase the biomass production. Final media composition consisted of glucose 25 g L^?1, yeast extract 5 g L^?1, Tween 80 0.38 % (v/v), Rice husk 10 g L^?1, CaCl₂ 1 g L^?1, and pH 4.88 ± 0.2. The Box-Behnken polynomial prediction resulted to be lower than the experimental validation of the model (6.59 vs. 6.91 Log₁₀ CFU ml^?1 respectively).     

Enhanced citric acid production by a yeast <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> over-expressing a pyruvate carboxylase gene

In this study, after the expression of a pyruvate carboxylase gene (PYC) cloned from Meyerozyma guilliermondii in a marine-derived yeast Yarrowia lipolytica SWJ-1b, a transformant PG86 obtained had much higher PYC activity than Y. lipolytica SWJ-1b. At the same time, the PYC gene expression and citric acid (CA) production by the transformant PG86 were also greatly enhanced. When glucose concentration in the medium was 60.0 g L^?1, CA concentration formed by the transformant PG86 was 34.02 g L^?1, leading to a CA yield of 0.57 g g^?1 of glucose. During a 10-L fed-batch fermentation, the final concentration of CA was 101.0 ± 1.3 g L^?1, the yield was 0.89 g g^?1 of glucose, the productivity was 0.42 g L^?1 h^?1 and only 5.93 g L^?1 reducing sugar was left in the fermented medium within 240 h of the fed-batch fermentation. HPLC analysis showed that most of the fermentation products were CA.     

The citric acid production from raw glycerol by Yarrowia lipolytica yeast and its regulation

The optimal cultivation conditions ensuring the maximal rate of citric acid (CA) biosynthesis by glycerol-grown mutant Yarrowia lipolytica NG40/UV7 were found to be as follows: growth limitation by inorganic nutrients (nitrogen, phosphorus, or sulfur), 28 °C, pH 5.0, dissolved oxygen concentration (pO₂) of 50 % (of air saturation), and pulsed addition of glycerol from 20 to 80 g L^?1 depending on the rate of medium titration. Under optimal conditions of fed-batch cultivation, in the medium with pure glycerol, strain Y. lipolytica NG40/UV7 produced 115 g L^?1 of CA with the mass yield coefficient of 0.64 g g^?1 and isocitric acid (ICA) amounted to 4.6 g L^?1; in the medium with raw glycerol, CA production was 112 g L^?1 with the mass yield coefficient of 0.90 g g^?1 and ICA amounted to 5.3 g L^?1. Based on the activities of enzymes involved in the initial stages of raw glycerol assimilation, the tricarboxylic acid cycle and the glyoxylate cycle, the mechanism of increased CA yield from glycerol-containing substrates in Y. lipolytica yeast was explained.     

Butanol production from concentrated lactose/whey permeate: Use of pervaporation membrane to recover and concentrate product

In these studies, butanol (acetone butanol ethanol or ABE) was produced from concentrated lactose/whey permeate containing 211 g L^?1 lactose. Fermentation of such a highly concentrated lactose solution was possible due to simultaneous product removal using a pervaporation membrane. In this system, a productivity of 0.43 g L^?1 h^?1 was obtained which is 307 % of that achieved in a non-product removal batch reactor (0.14 g L^?1 h^?1) where approximately 60 g L^?1 whey permeate lactose was fermented. The productivity obtained in this system is much higher than that achieved in other product removal systems (perstraction 0.21 g L^?1 h^?1 and gas stripping 0.32 g L^?1 h^?1). This membrane was also used to concentrate butanol from approximately 2.50 g L^?1 in the reactor to 755 g L^?1. Using this membrane, ABE selectivities and fluxes of 24.4–44.3 and 0.57–4.05 g m^?2 h^?1 were obtained, respectively. Pervaporation restricts removal of water from the reaction mixture thus requiring significantly less energy for product recovery when compared to gas stripping.     

Phosphorus retention of forested and emergent marsh depressional wetlands in differing land uses in Florida,USA

The translocation of phosphorus (P) from terrestrial landscapes to aquatic bodies is of concern due to the impact of elevated P on aquatic system functioning and integrity. Due to their common location in depressions within landscapes, wetlands, including so-called geographically isolated wetlands (GIWs), receive and process entrained P. The ability of depressional wetlands, or GIWs, to sequester P may vary by wetland type or by land use modality. In this study we quantified three measures of P sorption capacities for two common GIW types (i.e., emergent marsh and forested wetlands) in two different land use modalities (i.e., agricultural and least impacted land uses) across 55 sites in Florida, USA. The equilibrium P concentration (EPC₀) averaged 6.42 ± 5.18 mg P L^?1 (standard deviation reported throughout); and ranged from 0.01–27.18 mg P L^?1; there were no differences between GIW type or land use modality, nor interaction effects. Significant differences in phosphorus buffering capacity (PBC) were found between GIW types and land use, but no interaction effects. Forested GIWs [average 306.64 ± 229.63 (mg P kg^?1) (µg P L^?1)^?1], and GIWs in agricultural settings [average 269.95 ± 236.87 (mg P kg^?1) (µg P L^?1)^?1] had the highest PBC values. The maximum sorption capacity (S_max) was found to only differ by type, with forested wetlands (1274.5 ± 1315.7 mg P kg^?1) having over three times the capacity of emergent GIWs (417.5 ± 534.6 mg P kg^?1). Classification trees suggested GIW soil parameters of bulk density, organic content, and concentrations of total P, H₂O-extractable P, and HCl-extractable P were important to classifying GIW P-sorption metrics. We conclude that GIWs have high potential to retain P, but that the entrained P may be remobilized to the wetland water column depending on storm and groundwater input P concentrations. The relative hydrologic dis-connectivity of GIWs from other aquatic systems may provide sufficient retention time to retain elevated P within these systems, thereby providing an ecosystem service to downstream waters.