Enantioseparation of flurbiprofen enantiomers using chiral ionic liquids by liquid‐liquid extraction

Flurbiprofen is a kind of nonsteroidal anti‐inflammatory drug, which has been widely used in clinic for treatment of rheumatoid arthritis and osteoarthritis. It has been reported that S‐flurbiprofen shows good performance on clinic anti‐inflammatory treatment, while R‐enantiomer almost has no pharmacological activities. It has important practical values to obtain optically pure S‐flurbiprofen. In this work, chiral ionic liquids, which have good structural designability and chiral recognize ability, were selected as the extraction selector by the assistance of quantum chemistry calculations. The distribution behaviors of flurbiprofen enantiomers were investigated in the extraction system, which was composed of organic solvent and aqueous phase containing chiral ionic liquid. The results show that maximum enantioselectivity up to 1.20 was attained at pH 2.0, 25°C using 1,2‐dichloroethane as organic solvent, 1‐butyl‐3‐methylimidazole L‐tryptophan ([Bmim][L‐trp]) as chiral selector. The racemic flurbiprofen initial concentration was 0.2 mmol L^?1, and [Bmim][L‐trp] concentration was 0.02 mol L^?1. Furthermore, the recycle of chiral ionic liquids has been achieved by reverse extraction process of the aqueous phase with chiral selector, which is significant for industrial application of chiral ionic liquids and scale‐up of the extraction process.     

Evaluation of substrate promiscuity of an L‐carbamoyl amino acid amidohydrolase from Geobacillus stearothermophilus CECT43

N‐carbamoyl‐amino‐acid amidohydrolase (also known as N‐carbamoylase) is the stereospecific enzyme responsible for the chirality of the D ‐ or L ‐amino acid obtained in the “Hydantoinase Process.” This process is based on the dynamic kinetic resolution of D ,L ‐5‐monosubstituted hydantoins. In this work, we have demonstrated the capability of a recombinant L ‐N‐carbamoylase from the thermophilic bacterium Geobacillus stearothermophilus CECT43 (BsLcar) to hydrolyze N‐acetyl and N‐formyl‐L ‐amino acids as well as the known N‐carbamoyl‐L ‐amino acids, thus proving its substrate promiscuity. BsLcar showed faster hydrolysis for N‐formyl‐L ‐amino acids than for N‐carbamoyl and N‐acetyl‐L ‐derivatives, with a catalytic efficiency (k_cat/K_m) of 8.58 × 10⁵, 1.83 × 10⁴, and 1.78 × 10³ (s^?1 M^?1), respectively, for the three precursors of L ‐methionine. Optimum reaction conditions for BsLcar, using the three N‐substituted‐L ‐methionine substrates, were 65°C and pH 7.5. In all three cases, the metal ions Co²⁺, Mn²⁺, and Ni²⁺ greatly enhanced BsLcar activity, whereas metal‐chelating agents inhibited it, showing that BsLcar is a metalloenzyme. The Co²⁺‐dependent activity profile of the enzyme showed no detectable inhibition at high metal ion concentrations. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Absolute configuration and biological profile of pyrazoline enantiomers as MAO inhibitory activity

A new racemic pyrazoline derivative was synthesized and resolved to its enantiomers using analytic and semipreparative high‐pressure liquid chromatography. The absolute configuration of both fractions was established using vibrational circular dichroism. The in vitro monoamine oxidase (MAO) inhibitory profiles were evaluated for the racemate and both enantiomers separately for the two isoforms of the enzyme. The racemic compound and both enantiomers were found to inhibit hMAO‐A selectively and competitively. In particular, the R enantiomer was detected as an exceptionally potent and a selective MAO‐A inhibitor (K_i = 0.85 × 10^?3 ± 0.05 × 10^?3 μM and SI: 2.35 × 10^?5), whereas S was determined as poorer compound than R in terms of K_i and SI (0.184 ± 0.007 and 0.001). The selectivity of the enantiomers was explained by molecular modeling docking studies based on the PDB enzymatic models of MAO isoforms.     

Design and synthesis of molecular probes for the determination of the target of the anthelmintic drug praziquantel

Schistosomiasis is a highly prevalent neglected tropical disease caused by blood-dwelling helminths of the genus Schistosoma. Praziquantel (PZQ) is the only drug available widely for the treatment of this disease and is administered in racemic form, even though only the (R)-isomer has significant anthelmintic activity. Progress towards the development of a second generation of anthelmintics is hampered by a lack of understanding of the mechanism of action of PZQ. In this Letter, we report an efficient protocol for the small-scale separation of enantiomers of 2 (hydrolyzed PZQ) using supercritical fluid chromatography (SFC). The enantiopure 2 was then used to develop several molecular probes, which can potentially be used to help identify the protein target of PZQ and study its mode of action.     

Stereoselective effect of kynurenine enantiomers on the excretion of serotonin and its metabolite in rat urine

A solution of optically pure kynurenine (KYN), i.e., D ‐KYN or L ‐KYN, was administered intravenously to male Sprague‐Dawley rats (10 mg kg^?1 ml^?1). The time‐course of changes in the concentrations of urinary monoamines and their metabolites such as 5‐hydroxytryptamine (5‐HT), 5‐hydroxyindole acetic acid (5‐HIAA), dopamine, and 3‐methoxytyramine were investigated by reversed‐phase high‐performance liquid chromatography with electrochemical detection after precolumn derivatization with (2R)‐2,5‐dioxopyrrolidin‐1‐yl‐2,5,7,8‐tetramethyl‐6‐(tetrahydro‐2H‐pyran‐2‐yloxy)chroman‐2‐carboxylate (NPCA). We observed a stereoselective difference in the effects of the KYN enantiomers. Only D ‐KYN, not L ‐KYN, caused a significant increase in urinary 5‐HT levels within 30 min after its administration. With regard to the metabolites, urinary 3‐MT level was increased by D ‐KYN administration. On the other hand, no significant change in the DA level was observed after administration of either D ‐KYN or L ‐KYN. These results suggest that D ‐KYN could affect the activity of neuroactive amines, especially 5‐HT, in vivo. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors

Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in large‐scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D‐fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design‐of‐experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D‐fluorescence could more accurately measure ammonium concentration (RMSE_CV 0.031 g L^?1) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSE_CV 1.11 and 0.92 g L^?1, respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D‐fluorescence. The implementation of Raman spectroscopy increases at‐line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337–346, 2017     

Inhibitory effects of gossypol-related compounds on growth of Aspergillus flavus

Aims: The objective of this study was to test a series of gossypol‐related compounds for growth inhibition against Aspergillus flavus. Methods and Results: A series of chiral and achiral gossypol derivatives, some natural products of the cotton plant and others prepared by synthesis from gossypol, were incorporated into agar plates to follow the rate of A. flavus isolate AF13 colony growth. All tested compounds exhibited some growth inhibition against this organism. The synthetic compounds, gossypolone and apogossypolone, exhibited greater activity than either racemic or chiral gossypol. Methylated derivatives (i.e. 6‐methoxy and 6,6′‐dimethoxy derivatives) generally exhibited less activity than the nonmethylated parent compounds. The (?)‐optical form of gossypol was found to be slightly more active than the (+)‐optical form, and this trend was observed regardless of the presence of methoxy groups at the 6‐position. Growth inhibition of gossypolone and apogossypolone was concentration dependent. For gossypolone, the 50% effective dose was 90 μg ml^?1 of medium (165 μmol l^?1). For apogossypolone, the most active compound in the study, the 50% effective dose was 19 μg ml^?1 (38·7 μmol l^?1). The presence of gossypol‐related terpenoids appeared to stimulate production of A. flavus sclerotia, although replicate variability was so large that it was not possible to determine a significant correlation between the mass of sclerotia formed and compound growth inhibition. Conclusions: The quinone derivatives of gossypol, gossypolone and apogossypolone demonstrated significant fungal growth inhibitory activity against A. flavus. Significance and Impact of the Study: These gossypol derivatives may provide a new class of fungicide for use against the mycotoxigenic fungus A. flavus.     

Predicting soil carbon changes in switchgrass grown on marginal lands under climate change and adaptation strategies

The United States Great Lakes Region (USGLR) is a critical geographic area for future bioenergy production. Switchgrass (Panicum virgatum) is widely considered a carbon (C)‐neutral or C‐negative bioenergy production system, but projected increases in air temperature and precipitation due to climate change might substantially alter soil organic C (SOC) dynamics and storage in soils. This study examined long‐term SOC changes in switchgrass grown on marginal land in the USGLR under current and projected climate, predicted using a process‐based model (Systems Approach to Land‐Use Sustainability) extensively calibrated with a wealth of plant and soil measurements at nine experimental sites. Simulations indicate that these soils are likely a net C sink under switchgrass (average gain 0.87 Mg C ha^?1 year^?1), although substantial variation in the rate of SOC accumulation was predicted (range: 0.2–1.3 Mg C ha^?1 year^?1). Principal component analysis revealed that the predicted intersite variability in SOC sequestration was related in part to differences in climatic characteristics, and to a lesser extent, to heterogeneous soils. Although climate change impacts on switchgrass plant growth were predicted to be small (4%–6% decrease on average), the increased soil respiration was predicted to partially negate SOC accumulations down to 70% below historical rates in the most extreme scenarios. Increasing N fertilizer rate and decreasing harvest intensity both had modest SOC sequestration benefits under projected climate, whereas introducing genotypes better adapted to the longer growing seasons was a much more effective strategy. Best‐performing adaptation scenarios were able to offset >60% of the climate change impacts, leading to SOC sequestration 0.7 Mg C ha^?1 year^?1 under projected climate. On average, this was 0.3 Mg C ha^?1 year^?1 more C sequestered than the no adaptation baseline. These findings provide crucial knowledge needed to guide policy and operational management for maximizing SOC sequestration of future bioenergy production on marginal lands in the USGLR.     

Effects of irradiance, water temperature and nutrients on the growth of sporelings of the crustose coralline alga Lithophyllum yessoense Foslie (Corallinales, Rhodophyceae)

Lithophyllum yessoense Foslie is a markedly dominant subtidal, crustose coralline alga in south–western Hokkaido, Japan. In this study, the effects of irradiance, water temperature and nutrients (nitrate and phosphate) on the growth of sporelings of the alga were examined. The relative growth rate (RGR) was saturated at 17.6% d^?1 at a high irradiance (240 umol photon m²s^?1). Even at a low irradiance (10.7–49.9 umol photon m^?2s^?1), RGR was 7.1–12.7% d^?1 The survival rate of sporelings was greater than 80% at irradiance above 10.7 μmol photon m^?2s^?1 throughout the culture period. The growth of L. yessoense sporelings was promoted at 15°C and 20°C, but inhibited at 5°C. The half‐saturation constants (Ks) for growth were about 0.5 umol L^?1 and 0.14 umol L^?1 for nitrate and phosphate, respectively. Saturated nitrate and phosphate concentrations for the growth were about 4.0 μmol L^?1 and 0.4 μmol L^?1, respectively, suggesting that L. yessoense is adaptable to a relatively high water temperature, a wide range of irradiance, and low ambient nitrate and phosphate concentrations. The results provide a possible explanation of why L. yessoense is dominant in the environments of south‐western Hokkaido.     

Biorefinery for combined production of jet fuel and ethanol from lipid‐producing sugarcane: a techno‐economic evaluation

Replacing fossil fuels with an economically viable green alternative at scale has proved most challenging in the aviation sector. Recently sugarcane, the most productive crop on the planet, has been engineered to accumulate lipids. This opens the way for production of far more industrial vegetable oil per acre than previously possible. This study performs techno‐economic feasibility analysis of jet fuel production from this new cost efficient and high yield feedstock. A comprehensive process model for biorefinery producing hydrotreated jet fuel (from lipids) and ethanol (from sugars), with 1 600 000 MT yr^?1 lipid‐cane processing capacity, was developed in SuperPro Designer. Considering lipid‐cane development is continuing for higher oil concentrations, analysis was performed with lipid‐cane containing 5%, 10%, 15%, and 20% lipids. Capital investments for the biorefinery ranged from 238.1 to 351.2 million USD, with jet fuel capacities of 12.6–50.5 million liters (correspondingly ethanol production of nil to 102.6 million liters). The production cost of jet fuel for different scenarios was estimated Replacing fossil fuels with an economically viable green alternative at scale has proved most challenging in the aviation sector. Recently sugarcane, the most productive crop on the planet, has been engineered to accumulate lipids. This opens the way for production of far more industrial vegetable oil per acre than previously possible. This study performs techno‐economic feasibility analysis of jet fuel production from this new cost efficient and high yield feedstock. A comprehensive process model for biorefinery producing hydrotreated jet fuel (from lipids) and ethanol (from sugars), with 1 600 000 MT yr^?1 lipid‐cane processing capacity, was developed in SuperPro Designer. Considering lipid‐cane development is continuing for higher oil concentrations, analysis was performed with lipid‐cane containing 5%, 10%, 15%, and 20% lipids. Capital investments for the biorefinery ranged from 238.1 to 351.2 million USD, with jet fuel capacities of 12.6–50.5 million liters (correspondingly ethanol production of nil to 102.6 million liters). The production cost of jet fuel for different scenarios was estimated $0.73 to $1.79 per liter ($2.74 to $6.76 per gal) of jet fuel. In all cases, the cost of raw materials accounted for more than 70% of total operational cost. Biorefinery was observed self‐sustainable for steam and electricity requirement, because of in‐house steam and electricity generation from burning of bagasse. Minimum fuel selling prices with a 10% discount rate for 20% lipid case was estimated $1.40/L ($5.31/gal), which was lower than most of the reported prices of renewable jet fuel produced from other oil crops and algae. Along with lower production costs, lipid‐cane could produce as high as 16 times the jet fuel (6307 L ha^?1) per unit land than that of other oil crops and do so using low‐value land unsuited to most other crops, while being highly water and nitrogen use efficient.     

Nisin production of Lactococcus lactis N8 with hemin‐stimulated cell respiration in fed‐batch fermentation system

In this study, nisin production of Lactococcus lactis N8 was optimized by independent variables of glucose, hemin and oxygen concentrations in fed‐batch fermentation in which respiration of cells was stimulated with hemin. Response surface model was able to explain the changes of the nisin production of L. lactis N8 in fed‐batch fermentation system with high fidelity (R² 98%) and insignificant lack of fit. Accordingly, the equation developed indicated the optimum parameters for glucose, hemin, and dissolved oxygen were 8 g L^?1 h^?1, 3 μg mL^?1 and 40%, respectively. While 1711 IU mL^?1 nisin was produced by L. lactis N8 in control fed‐batch fermentation, 5410 IU mL^?1 nisin production was achieved within the relevant optimum parameters where the respiration of cell was stimulated with hemin. Accordingly, nisin production was enhanced 3.1 fold in fed‐batch fermentation using hemin. In conclusion the nisin production of L. lactis N8 was enhanced extensively as a result of increasing the biomass by stimulating the cell respiration with adding the hemin in the fed‐batch fermentation. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:678–685, 2015     

Selective sensing Ca2+ with a spiropyran‐based fluorometric probe

Spiropyran (SP) and its derivatives operate between their ring opening and closing forms as a versatile molecular platform for the fluorescence detection of cations and anions, using a colour change for signalling. A functionalized SP fluorescence probe, L , was prepared and characterized. Probe L can detect Ca²⁺ with a fluorescence ‘turn‐on’ response in ethanol solution. It selectively binds Ca²⁺ to form a 1:1 ligand/metal complex, which produced a new emission band centred at 604 nm. The sensing result was clearly observed by the solution colour change from colourless to pink under visible light, and from blue to red under ultraviolet light. The detection limit was calculated to be 4.53 × 10^?8 M for Ca²⁺. The probe provides another possibility that SP‐based derivatives could be used for the development and detection of metal ions in environmental and physiological systems.     

THE PERFORMANCE OF SPRAY‐IRRIGATED ULVA LACTUCA (ULVOPHYCEAE,CHLOROPHYTA) AS A CROP AND AS A BIOFILTER OF FISHPOND EFFLUENTS1

The seaweed Ulva lactuca L. was spray cultured by mariculture effluents in a mattress‐like layer, held in air on slanted boards by plastic netting. Air‐agitated seaweed suspension tanks were the reference. Growth rate, yield, and ammonia‐N removal rate were 11.8% · d^?1, 171 g fresh weight (fwt) · m^?2 · d^?1, and 5 g N · m^?2 · d^?1, respectively, by the spray‐cultured U. lactuca, and 16.9% · d^?1, 283 g fwt · m^?2 · d^?1, and 7 g N · m^?2 · d^?1, respectively, by the tank U. lactuca. Biomass protein content was similar in both treatments. Dissolved oxygen in the fishpond effluent water was raised by >3 mg · L^?1 and pH by up to half a unit, upon passage through both culture systems. The data suggest that spray‐irrigation culture of U. lactuca in this simple green‐mattress‐like system supplies the seaweed all it needs to grow and biofilter at rates close to those in standard air‐agitated tank culture.     

Taste,A New Incentive to Switch to (R)-Praziquantel in Schistosomiasis Treatment

Background

Praziquantel (PZQ) is the drug compound of choice in the control and treatment of schistosomiasis. PZQ is administered as a racemate, i. e. 1∶1 mixture of enantiomers. The schistosomicidal activity arises from one PZQ-enantiomer, whereas the other enantiomer does not contribute to the activity. The WHO''s Special Programme for Research and Training in Tropical Diseases (TDR) has assigned the low-cost preparation of pure schistosomicidal (−)-PZQ a key priority for future R&D on PZQ, but so far this transition has not happened. PZQ has two major administration drawbacks, the first being the high dose needed, and its well documented bitter and disgusting taste. Attempts of taste-masking by low-cost means have not been successful. We hypothesized that the non-schistosomicidal component in PZQ would be the main contributor to the unpleasant taste of the drug. If the hypothesis was confirmed, the two major administration drawbacks of PZQ, the high dose needed and its bitter taste, could be addressed in one go by removing the component contributing to the bitter taste.

Methods and Findings

PZQ was separated into its schistosomicidal and the non-schistosomicidal component, the absolute stereochemical configuration of (−)-PZQ was determined to be (R)-PZQ by X-ray crystallography, and the extent of bitterness was determined for regular racemic PZQ and the schistosomicidal component in a taste study in humans. Finding: The schistosomicidal component alone is significantly less bitter than regular, racemic PZQ.

Conclusion

Our hypothesis is confirmed. We propose to use only the pure schistosomicidal component of PZQ, offering the advantage of halving the dose and expectedly improving the compliance due to the removal of the bitter taste. Therefore, (R)-PZQ should be specifically suitable for the treatment of school-age children against schistosomiasis. With this finding, we would like to offer an additional incentive to the TDR''s recommendation to switch to the pure schistosomicidal (R)-PZQ.     

Metabolism in 1,3‐propanediol fed‐batch fermentation by a D‐lactate deficient mutant of Klebsiella pneumoniae

Klebsiella pneumoniae HR526, a new isolated 1,3‐propanediol (1,3‐PD) producer, exhibited great productivity. However, the accumulation of lactate in the late‐exponential phase remained an obstacle of 1,3‐PD industrial scale production. Hereby, mutants lacking D ‐lactate pathway were constructed by knocking out the ldhA gene encoding fermentative D ‐lactate dehydrogenase (LDH) of HR526. The mutant K. pneumoniae LDH526 with the lowest LDH activity was studied in aerobic fed‐batch fermentation. In experiments using pure glycerol as feedstock, the 1,3‐PD concentrations, conversion, and productivity increased from 95.39 g L^?1, 0.48 and 1.98 g L^?1 h^?1 to 102. 06 g L^?1, 0.52 mol mol^?1 and 2.13 g L^?1 h^?1, respectively. The diol (1,3‐PD and 2,3‐butanediol) conversion increased from 0.55 mol mol^?1 to a maximum of 0.65 mol mol^?1. Lactate would not accumulate until 1,3‐PD exceeded 84 g L^?1, and the final lactate concentration decreased dramatically from more than 40 g L^?1 to <3 g L^?1. Enzymic measurements showed LDH activity decreased by 89–98% during fed‐batch fermentation, and other related enzyme activities were not affected. NADH/NAD⁺ enhanced more than 50% in the late‐exponential phase as the D ‐lactate pathway was cut off, which might be the main reason for the change of final metabolites concentrations. The ability to utilize crude glycerol from biodiesel process and great genetic stability demonstrated that K. pnemoniae LDH526 was valuable for 1,3‐PD industrial production. Biotechnol. Bioeng. 2009; 104: 965–972. © 2009 Wiley Periodicals, Inc.     

Exploring aggregation‐induced emission through tuning of ligand structure for picomolar detection of pyrene

Tuning of ligand structures through controlled variation of ring number in fused‐ring aromatic moiety appended to antipyrine allows detection of 7.8 × 10^?12 M pyrene via aggregation‐induced emission (AIE) associated with 101‐fold fluorescence enhancement. In one case, antipyrine unit is replaced by pyridine to derive bis‐methylanthracenyl picolyl amine. The structures of four molecules have been confirmed by single crystal X‐ray diffraction analysis. Among them, pyrene‐antipyrine conjugate (L) undergoes pyrene triggered inhibition of photo‐induced electron transfer (PET) leading to water‐assisted AIE.     

Detection of Xeljanz Enantiomers in Diethyl Amine Active Pharmaceutical Ingredients and Tablets

A high‐performance liquid chromatography (HPLC) method was established to detect Xeljanz enantiomers in active pharmaceutical ingredients (APIs) and tablets. The separation was achieved on a Chiralpak IC column using a mobile phase of hexane‐ethanol‐diethylamine (65:35:0.1, v/v). The detection wavelength was 289 nm. The peak areas and the enantiomer concentrations in the range of 0.15–2.25 μg?mL^?1 were in high linearity, with correlation coefficients higher than 0.999. The recoveries were 86.44% at the concentrations of 7.5, 18.75, and 37.5 μg?mL^?1. The limit of detection (LOD) and limit of quantification (LOQ) were 0.042 and 0.14 μg?mL^?1, respectively. This HPLC method is suitable for detecting the enantiomers of Xeljanz in its APIs and tablets. Chirality 27:235–238, 2015. © 2014 Wiley Periodicals, Inc.     

Projections of the availability and cost of residues from agriculture and forestry

By‐products of agricultural and forestry processes, known as residues, may act as a primary source of renewable energy. Studies assessing the availability of this resource offer little insight on the drivers and constraints of the available potential as well as the associated costs and how these may vary across scenarios. This study projects long‐term global supply curves of the available potential using consistent scenarios of agriculture and forestry production, livestock production and fuel use from the spatially explicit integrated assessment model IMAGE. In the projections, residue production is related to agricultural and forestry production and intensification, and the limiting effect of ecological and alternative uses of residues are accounted for. Depending on the scenario, theoretical potential is projected to increase from approximately 120 EJ yr^?1 today to 140–170 EJ yr^?1 by 2100, coming mostly from agricultural production. To maintain ecological functions approximately 40% is required to remain in the field, and a further 20–30% is diverted towards alternative uses. Of the remaining potential (approximately 50 EJ yr^?1 in 2100), more than 90% is available at costs <10By‐products of agricultural and forestry processes, known as residues, may act as a primary source of renewable energy. Studies assessing the availability of this resource offer little insight on the drivers and constraints of the available potential as well as the associated costs and how these may vary across scenarios. This study projects long‐term global supply curves of the available potential using consistent scenarios of agriculture and forestry production, livestock production and fuel use from the spatially explicit integrated assessment model IMAGE. In the projections, residue production is related to agricultural and forestry production and intensification, and the limiting effect of ecological and alternative uses of residues are accounted for. Depending on the scenario, theoretical potential is projected to increase from approximately 120 EJ yr^?1 today to 140–170 EJ yr^?1 by 2100, coming mostly from agricultural production. To maintain ecological functions approximately 40% is required to remain in the field, and a further 20–30% is diverted towards alternative uses. Of the remaining potential (approximately 50 EJ yr^?1 in 2100), more than 90% is available at costs <10$₂₀₀₅ GJ^?1. Crop yield improvements increase residue productivity, albeit at a lower rate. The consequent decrease in agricultural land results in a lower requirement of residues for erosion control. The theoretical potential is most sensitive to baseline projections of agriculture and forestry demand; however, this does not necessarily affect the available potential which is relatively constant across scenarios. The most important limiting factors are the alternative uses. Asia and North America account for two‐thirds of the available potential due to the production of crops with high residue yields and socioeconomic conditions which limit alternative uses.     

Spectroscopic studies of ligand perturbation effects on the half oxidized action site of Busycon canaliculatum hemocyanin.

We report the preparation and characterization of an important new series of hemocyanin derivatives containing a variety of anions bound to the cupric ion in a singly oxidized active site. Spectroscopic studies on these half met-L derivatives, Cu(I)…Cu(II)-L, where L = CN^?, NO₂^?, CH₃CO₂^?, and N₃^? reveal that the unpaired electron is localized on one copper in an approximately tetragonal site with a d_{x² ? y²} ground state. Ligand variation is used as a perturbation on the spectral features to show that the small molecule (and by analogy, oxygen in oxyhemocyanin) is bound in the equatorial plane of the copper.     

Easy Access to Enantiomerically Pure Nonproteinogenic Amino Acids

The protease from Bacillus licheniformis (alcalase) shows a remarkable broad substrate tolerance and high enantioselectivity against nonproteinogenic racemic amino acid derivatives. N‐acetyl protected amino acid esters of mono‐, di‐ or tri‐substituted phenyl alanines and even tert.‐leucine were hydrolyzed with high enantioselectivity. The obtained mixtures of (S)‐N‐acetyl amino acid and (R)‐N‐acetyl amino acid ester can easily be separated. The R‐ or S‐amino acids were obtained by acidic cleavage of the optically pure derivatives or the (R)‐ester was racemized by treatment with potassium t‐butylate.