Discovery and Engineering of a Novel Baeyer-Villiger Monooxygenase with High Normal Regioselectivity

Baeyer-Villiger monooxygenases (BVMOs) are remarkable biocatalysts for the Baeyer-Villiger oxidation of ketones to generate esters or lactones. The regioselectivity of BVMOs is essential for determining the ratio of the two regioisomeric products (“normal” and “abnormal”) when catalyzing asymmetric ketone substrates. Starting from a known normal-preferring BVMO sequence from Pseudomonas putida KT2440 (PpBVMO), a novel BVMO from Gordonia sihwensis (GsBVMO) with higher normal regioselectivity (up to 97/3) was identified. Furthermore, protein engineering increased the specificity constant (k_cat/K_M) 8.9-fold to 484 s⁻¹ mM⁻¹ for 10-ketostearic acid derived from oleic acid. Consequently, by using the variant GsBVMO_C308L as an efficient biocatalyst, 10-ketostearic acid was efficiently transformed into 9-(nonanoyloxy)nonanoic acid, with a space-time yield of 60.5 g L⁻¹ d⁻¹. This study showed that the mutant with higher regioselectivity and catalytic efficiency could be applied to prepare medium-chain ω-hydroxy fatty acids through biotransformation of long-chain aliphatic keto acids derived from renewable plant oils.     

Controlling the Regioselectivity of Baeyer–Villiger Monooxygenases by Mutation of Active‐Site Residues

Baeyer–Villiger monooxygenase (BVMO)‐mediated regiodivergent conversions of asymmetric ketones can lead to the formation of “normal” or “abnormal” lactones. In a previous study, we were able to change the regioselectivity of a BVMO by mutation of the active‐site residues to smaller amino acids, which thus created more space. In this study, we demonstrate that this method can also be used for other BVMO/substrate combinations. We investigated the regioselectivity of 2‐oxo‐Δ³‐4,5,5‐trimethylcyclopentenylacetyl‐CoA monooxygenase from Pseudomonas putida (OTEMO) for cis‐bicyclo[3.2.0]hept‐2‐en‐6‐one ( 1 ) and trans‐dihydrocarvone ( 2 ), and we were able to switch the regioselectivity of this enzyme for one of the substrate enantiomers. The OTEMO wild‐type enzyme converted (?)‐ 1 into an equal (50:50) mixture of the normal and abnormal products. The F255A/F443V variant produced 90 % of the normal product, whereas the W501V variant formed up to 98 % of the abnormal product. OTEMO F255A exclusively produced the normal lactone from (+)‐ 2 , whereas the wild‐type enzyme was selective for the production of the abnormal product. The positions of these amino acids were equivalent to those mutated in the cyclohexanone monooxygenases from Arthrobacter sp. and Acinetobacter sp. (CHMO_Arthro and CHMO_Acineto) to switch their regioselectivity towards (+)‐ 2 , which suggests that there are hot spots in the active site of BVMOs that can be targeted with the aim to change the regioselectivity.     

Kinetics of glucoamylase hydrolysis of corn starch

Corn starch was hydrolysed by glucoamylase (EC 3.2.1.3, 1,4‐a‐D ‐glucan glucohydrolase) in a chemostat; 106 g L^?1 reducing sugar was produced at 40 °C after 46 h, whereas only 42 g L^?1 was produced at 30 °C. When corn starch concentration was increased from 64.6 g L^?1 to 161.5 g L^?1, the amount of reducing sugar produced increased with the initial substrate concentration. The studies show that there is a product concentration (p₀) above which product inhibition becomes apparent. A new model for starch hydrolysis considering the product inhibition point is presented. Copyright © 2006 Society of Chemical Industry     

Time‐dependent dosing of Fe2+ for improved lipopeptide production by marine Bacillus megaterium

BACKGROUND: Lipopeptide production is strongly influenced by trace metals. The availability of free Fe²⁺ in the media throughout the process of fermentation was found to be very critical. Since free Fe²⁺ was reported to be sequestered by the lipopeptide as it was produced, intermittent feeding of Fe²⁺ was strategized and optimized for enhanced lipopeptide production by marine Bacillus megaterium in glucose mineral salts medium (GMSM). RESULTS: Studies with the single‐dose Fe²⁺ (0.48 mmol L^?1) supplementation after 8 h of fermentation resulted in lipopeptide concentration of 3.3 ± 0.1 g L^?1. Lipopeptide production was further enhanced to 4.2 ± 0.15 g L^?1 by adopting a multi‐dose Fe²⁺ feeding strategy. The maximum product yield (Y_P/S) of 0.24 ± 0.02 g g^?1 with specific product formation rate (q_p) of 0.124 ± 0.01 g g^?1 h^?1 was achieved when 0.48 mmol L^?1 Fe²⁺ was fed intermittently at different times as per the designed strategy. CONCLUSION: Lipopeptide concentration was improved 4.7‐fold by single‐dosing and 5.8‐fold by multiple dosing of Fe²⁺, when compared with GMSM without Fe²⁺ supplementation. Copyright © 2012 Society of Chemical Industry     

Continuous biotransformation of pyrogallol to purpurogallin using cross‐linked enzyme crystals of laccase as catalyst in a packed‐bed reactor

Cross‐linked enzyme crystals (CLEC) of laccase were prepared by crystallizing laccase with 75% (NH₄)₂SO₄ and cross‐linking using 1.5% glutaraldehyde. The cross‐linked enzyme crystals were further coated with 1 mmol L^?1 β‐cyclodextrin by lyophilization. The lyophilized enzyme crystals were used as such for the biotransformation of pyrogallol to purpurogallin in a packed‐bed reactor. The maximum conversion (76.28%) was obtained with 3 mmol L^?1 pyrogallol at a residence time of 7.1 s. The maximum productivity (269.03 g L^?1 h^?1) of purpurogallin was obtained with 5 mmol L^?1 pyrogallol at a residence time of 3.5 s. The productivity was found to be 261.14 g L^?1 h^?1 and 251.1 g L^?1 h^?1 when concentrations of 3 mmol L^?1 and 7 mmol L^?1 respectively were used. The reaction rate of purpurogallin synthesis was maximum (2241.94 mg purpurogallin mg^?1 CLEC h^?1) at a residence time of 3.5 s, when 5 mmol L^?1 pyrogallol was used as the substrate. The catalyst to product ratio calculated for the present biotransformation was 1:2241. The CLEC laccase had very high stability in reuse and even after 650 h of continuous use, the enzyme did not lose its activity. Copyright © 2006 Society of Chemical Industry     

Pre‐treatment and utilization of raw glycerol from sunflower oil biodiesel for growth and 1,3‐propanediol production by Clostridium butyricum

BACKGROUND: The objective of the present work is to report an efficient pre‐treatment process for sunflower oil biodiesel raw glycerol (SOB‐RG) and its fermentation to 1,3‐propanediol. RESULTS: The growth inhibition percentages of Clostridium butyricum DSM 5431 on grade A (pH 4.0) and grade B (pH 5.0) phosphoric acid‐treated SOB‐RG were similar to those of pure glycerol at 20 g glycerol L^?1; i.e., 18.5 ± 0.707% to 20.5 ± 0.7% inhibition. In grade A, growth inhibition was reduced from 85.25 ± 0.35% to 32 ± 1.4% (a 53.25% reduction) at 40 g glycerol L^?1 by washing grade A raw glycerol twice with n‐hexanol (grade A‐2). The kinetic parameters for product formation and substrate consumption in anaerobic batch cultures gave almost similar values at 20 g glycerol L^?1, while at 50 g glycerol L^?1 volumetric productivity (Q_p) and specific rate of 1,3‐propanediol formation (q_p) were improved from 1.13 to 1.85 g L^?1 h^?1 and 1.60 to 2.65 g g^?1 h^?1, respectively, by employing grade A‐2 raw glycerol, while the yields were similar (0.5–0.52 g g^?1). CONCLUSION: The results are important as the pre‐treatment of SOB‐RG is necessary to develop bioprocess technologies for conversion of SOB‐RG to 1,3‐propanediol. Copyright © 2008 Society of Chemical Industry     

Optimization of the culture conditions for production of glutamate decarboxylase by Streptococcus salivarius ssp. thermophilus

The culture conditions for glutamate decarboxylase (GAD) production under submerged fermentation by Streptococcus salivarius ssp. thermophilus were investigated. The results indicated the optimum culture medium was composed as follows: 15.0 g L^?1 of peptone, 12.5 g L^?1 of beef extract, 12.5 g L^?1 of sucrose, 1.03 g L^?1 of dipotassium hydrogen phosphate, 5 g L^?1 of sodium acetate, 2 g L^?1 of ammonium dibasic citrate, 2.12 g L^?1 of calcium chloride, 1 g L^?1 of Tween 80, and initial pH 6.79. The optimum culture temperature and time were 37 °C and 12 h, respectively. Under these conditions, GAD production was 257.46 ± 5.12 U, which was about 1.45‐fold that of Man–Rogosa–Sharpe broth. Copyright © 2008 Society of Chemical Industry     

Inulin as a Promising Alternative Feedstock for Docosahexaenoic Acid Production by Schizochytrium sp. ATCC 20888

Schizochytrium sp. is considered as a promising alternative commercial source of docosahexaenoic acid (DHA), but the production is hindered by the high feedstock cost. In this study, inulin is used as a cheap and readily available feedstock for Schizochytrium sp. ATCC 20888 to produce DHA. The strain could not utilize inulin directly and therefore inulin first needed to be hydrolyzed. Compared with the acidic hydrolyzate by HCl and hydrolyzate by endo‐inulinase, the hydrolyzate by exo‐inulinase serves as the most effective carbon source for microalgal growth. Hydrolysis of inulin by exo‐inulinase is further optimized, and up to 97.8% of inulin conversion is obtained under the optimal conditions of 40 °C, pH 7.0, substrate concentration of 80 g L^?1 and exo‐inulinase loading of 2 g kg^?1 substrate for 12 h. The resulting hydrolyzate containing mainly fructose is used for the DHA production by the microalga. The lipid content in biomass, DHA content in total fatty acids, DHA yield, and DHA productivity at 72 h reach 45.26%, 35.59%, 5.64 g L^?1 and 1.88 g L^?1 d^?1, respectively. The results suggest that inulin is an excellent feedstock for Schizochytrium sp. suitable for commercial DHA production. Practical Applications: DHA is an essential nutrient for human health and is widely used in infant formula and functional food. As a reserve carbohydrate, inulin present in plants represents a cheap, abundant, and readily available bioresource. This study describes the suitability of inulin as a promising alternative to glucose for DHA production by Schizochytrium sp. Hence, a practical bioprocess for commercial DHA production from inulin by Schizochytrium sp. could be developed. As far as it is known, this is the first report of inulin as a feedstock for Schizochytrium sp. to produce DHA.     

Highly efficient synthesis of 5‐cyanovaleramide by Rhodococcus ruber CGMCC3090 resting cells

BACKGROUND: The chemo‐selective biocatalytic hydrolysis of nitriles presents a valuable alternative to chemical hydrolysis, a procedure in which harsh conditions are applied. In this study, Rhodococcus ruber CGMCC3090 is used for 5‐cyanovaleramide (5‐CVAM) production by means of adiponitrile (ADN) hydration. Several parameters that affect the biocatalyzation process are investigated. RESULTS: The effective production of 5‐CVAM from ADN with good regioselectivity was successfully achieved using the resting cells of Rhodococcus ruber CGMCC3090. The reaction parameters for 5‐CVAM production were investigated during the experiment, and it was found that resting cells were effective in converting ADN at high concentrations (up to 2.0 mol L^?1) and the hydration reaction was slightly inhibited by different concentrations of 5‐CVAM. After 100 min of incubation, more than 99.2% of the added adiponitrile was converted to 5‐CVAM when the cell concentration was 1.05 g dry cell weight (DCW) L^?1. CONCLUSION: The study provides a facile and feasible way of achieving high 5‐CVAM production levels while maintaining high purity of the product obtained. Copyright © 2012 Society of Chemical Industry     

Conversion efficiency of glucose/xylose mixtures for ethanol production using Saccharomyces cerevisiae ITV01 and Pichia stipitis NRRL Y‐7124

BACKGROUND: Efficient conversion of glucose/xylose mixtures from lignocellulose is necessary for commercially viable ethanol production. Oxygen and carbon sources are of paramount importance for ethanol yield. The aim of this work was to evaluate different glucose/xylose mixtures for ethanol production using S. cerevisiae ITV‐01 (wild type yeast) and P. stipitis NRRL Y‐7124 and the effect of supplying oxygen in separate and co‐culture processes. RESULTS: The complete conversion of a glucose/xylose mixture (75/30 g L^?1) was obtained using P. stipitis NRRL Y‐7124 under aerobic conditions (0.6 vvm), the highest yield production being Y_p/s = 0.46 g g^?1, volumetric ethanol productivity Q_pmax = 0.24 g L^?1 h^?1 and maximum ethanol concentration P_max = 34.5 g L^?1. In the co‐culture process and under aerobic conditions, incomplete conversion of glucose/xylose mixture was observed (20.4% residual xylose), with a maximum ethanol production of 30.3 g L^?1, ethanol yield of 0.4 g g^?1 and Q_pmax = 1.26 g L^?1 h^?1. CONCLUSIONS: The oxygen present in the glucose/xylose mixture promotes complete sugar consumption by P. stipitis NRRL Y‐7124 resulting in ethanol production. However, in co‐culture with S. cerevisiae ITV‐01 under aerobic conditions, incomplete fermentation occurs that could be caused by oxygen limitation and ethanol inhibition by P. stipitis NRRL Y‐7124; nevertheless the volumetric ethanol productivity increases fivefold compared with separate culture. Copyright © 2011 Society of Chemical Industry     

Effect of oxygen transfer rates on alcohols production by Candida guilliermondii cultivated on soybean hull hydrolysate

BACKGROUND: In this research the use of soybean hull hydrolysate (SHH) as substrate for xylitol and ethanol production using an osmotolerant strain of Candida guilliermondii was studied. The production of alcohols was investigated in batch cultivations in which the variable parameter was the volumetric oxygen mass transfer coefficient (k_La) obtained from three different conditions of air supply: anaerobic (150 rpm, no aeration); microaerobic (300 rpm, 1 vvm), and aerobic (600 rpm, 2 vvm), corresponding to k_La values of 0; 8; and 46 h^?1, respectively. RESULTS: SHH, although presenting a very high osmotic pressure (1413 mOsm kg^?1), was completely metabolized under aerobic conditions with high biomass productivities of 0.49 g cells (L h)^?1, with little formation of ethanol. Xylitol was produced under microaeration, with product yield of 0.22 g g^?1 xylose, with the formation of glycerol as a by‐product. No xylose was metabolized under anaerobic conditions, but ethanol was produced from hexoses with high product yields of 0.5 g g^?1. CONCLUSION: These results suggest that the hydrolysis of soybean hull and its conversion to ethanol and other alcohols could be an important use of this agro‐industrial waste, which could be used for biofuel, xylitol or biomass production, depending on the aeration conditions of the cultures. Copyright © 2008 Society of Chemical Industry     

Production of L‐lactic acid with very high gravity distillery wastewater from ethanol fermentation by a newly isolated Enterococcus hawaiiensis

BACKGROUND: A great amount of wastewater with high contents of chemical oxygen demand (COD) are produced by ethanol production. It would be useful to utilize distillery wastewater to produce L‐lactic acid, which could be a high additional value byproduct of ethanol production. The fermentation process of L‐lactic acid production by a newly isolated Enterococcus hawaiiensis CICIM‐CU B0114 is reported for the first time. RESULTS: The strain produced 56 g L^?1 of L‐lactic acid after cultivation for 48 h in optimized medium consisting of (g L^?1) 80 glucose, 10 peptone, 10 yeast extract, 1.5 Na₂HPO₄ and 0.2 MgSO₄. E. hawaiiensis CICIM‐CU B0114 was isolated and purified by subculture for growing and producing L‐lactic acid in distillery wastewater of very high gravity (VHG) from ethanol fermentation. L‐lactic acid fermentation was further studied with distillery wastewater substrate in 7 L and 15 L fermentors. The results showed that L‐lactic acid concentrations of 52 g L^?1 and 68 g L^?1 was achieved in 7 L and 15 L fermentors with the initial sugar concentrations of 67 g L^?1 and 87 g L^?1, respectively. CONCLUSION: The production of L‐lactic acid by the newly isolated E. hawaiiensis CICIM‐CU B0114 was carried out and the fermentation medium was optimized by orthogonal experimental design. This new strain holds the promise of L‐lactic acid production utilizing distillery wastewater from VHG ethanol fermentation. Copyright © 2010 Society of Chemical Industry     

Multiobjective optimization of microalgae (Chlorella sp.) growth in a photobioreactor using Box‐Behnken design approach

This study investigates a parameter optimization approach to maximize the specific growth rate of the Chlorella vulgaris microalgae species, its biomass productivity, and CO₂ capture rate. For this purpose, the Box‐Behnken experimental design technique is applied with temperature, nitrogen to phosphorus ratio, and light‐dark cycle per day, as the growth controlling parameters. For each response, a quadratic model is developed separately describing the algal specific growth rate, biomass productivity, and CO₂ capture rate, respectively. The maximum specific growth rate of 0.84 d^?1 is obtained at 25 °C, with a nitrogen to phosphorus ratio of 3.4:1, and light‐dark cycles of 24/0 h. Maximum biomass productivity of 147.3 mg L^?1 d^?1 is found at 30 °C, with a nitrogen to phosphorus ratio of 3:1, and light‐dark cycles of 12/12 h. In addition, the maximum CO₂ capture rate of 159.5 mg L^?1 d^?1 is also obtained at 30 °C, with a nitrogen to phosphorus ratio of 4:1, and light‐dark cycles of 23/1 h. Finally, a multi‐response optimization method is applied to maximize the specific growth rate, biomass productivity, and CO₂ capture rate, simultaneously. The optimal set of 30 °C, a nitrogen to phosphorus ratio 3:1, and light‐dark cycles 16/8 h, provide the maximum specific growth rate of 0.66 per day, biomass productivity of 147.6 mg L^?1 d^?1, and CO₂ capture rate of 141.7 mg L^?1 d^?1.

     

Novel two‐in‐one bioreactor greatly improves lactic acid production from xylose by Lactobacillus pentosus

BACKGROUND: Simultaneous xylose isomerization and fermentation was investigated to improve the lactic acid production from xylose by Lactobacillus pentosus in a novel two‐in‐one bioreactor constructed by packing the immobilized xylose isomerase (65 g) in a fixed bed reactor (diameter 56 mm × 66 mm, packing volume 154 mL) with a permeable wall, which was installed inside a conventional fermenter (2 L) and rotated along the axis together with the mechanical stirrer of the fermenter. RESULTS: Xylose (20 g L^?1) was completely consumed within 24 h in the novel bioreactor, compared with 72 h needed for the control without packed enzyme. The maximum cell density (17.5 g L^?1) in the novel bioreactor was twice that in the control and the lactic acid productivity (0.58 g L^?1 h^?1) was 3.8 times higher. Repeated use of the immobilized enzyme showed that the lactic acid productivity and yield obviously dropped after the first batch fermentation but maintained almost unchanged afterwards. CONCLUSION: Simultaneous xylose isomerization and fermentation significantly improved lactic acid production from xylose by Lactobacillus pentosus. The novel bioreactor made it easier to recycle and reuse the immobilized enzyme. © 2012 Society of Chemical Industry     

Fermentative production of L(+)‐lactic acid from starch hydrolyzate and corn steep liquor as inexpensive nutrients by batch culture of Enterococcus faecalis RKY1

BACKGROUND: Attempts were made to determine the lactic acid production efficiency of novel isolate, Enterococcus faecalis RKY1 using four different starches (corn, tapioca, potato, and wheat starch) with different concentrations (50, 75, 100, and 125 g L^?1) and corn steep liquor as an inexpensive nitrogen source. RESULTS: The yield of lactic acid from each starch was higher than 95% based on initial starch concentrations. High lactic acid concentration (129.9 g L^?1) and yield (1.04 g‐lactic acid g^?1‐starch) were achieved faster (84 h) from 125 g L^?1 of corn starch. Among the starches used, tapioca starch fermentation usually completed in a shorter incubation period. The final dry cell weight was highest (7.0 g L^?1) for the medium containing 75 g L^?1 of corn starch, which resulted in maximum volumetric productivity of lactic acid (3.6 g L^?1 h^?1). The addition of 30 g L^?1 corn steep liquor supplemented with a minimal amount of yeast extract supported both cell growth and lactic acid fermentation. CONCLUSION: Enterococcus faecalis RKY1 was found to be capable of growing well on inexpensive nutrients and producing maximum lactic acid from starches and corn steep liquor as lower‐cost raw materials than conventionally‐used refined sugars such as glucose, and yeast extract as an organic nitrogen source in laboratory‐scale studies. These fermentation characteristics are prerequisites for the industrial scale production of lactic acid. Copyright © 2008 Society of Chemical Industry     

Production of organic solvent tolerant lipase by Staphylococcus caseolyticus EX17 using raw glycerol as substrate

BACKGROUND: In this work we used Plackett–Burman statistical design and central composite design in order to optimize culture conditions for lipase production by Staphylococcus caseolyticus strain EX17 growing on raw glycerol, which was obtained as a by‐product of the enzymatic synthesis of biodiesel. The stability of lipase was verified over several organic solvents, such as methanol, ethanol and n‐hexane. RESULTS: Optimal culture conditions for lipase production were found to be 36 °C, initial pH 8.12, glycerol 30 g L^?1, olive oil 3.0 g L^?1, and soybean oil 2.5 g L^?1, with 145.8 U L^?1 of enzyme activity. When commercial glycerol was substituted by the raw glycerol from biodiesel synthesis, lipolytic activity was 127.3 U L^?1. Experimental validation of enzyme production matched values predicted by the mathematical model, which was 138.3 U L^?1. Stability tests showed that lipase from S. caseolyticus EX17 was stable in methanol, ethanol, and n‐hexane. CONCLUSIONS: Results obtained in this work suggest that raw glycerol can be used for lipase production by S. caseolyticus EX17 and that this enzyme has a potential application in the synthesis of biodiesel. Copyright © 2008 Society of Chemical Industry     

Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two‐stage fed‐batch fermentation process

BACKGROUND: Xylitol, a sugar alcohol widely used in food and pharmaceutical industries, can be produced through biological reduction of xylose present in hemicellulose hydrolysates by Candida tropicalis. However, the aeration rate and by‐products originating from hemicellulose hydrolysis strongly inhibit the production of xylitol in a fermentation process. A two‐stage fed‐batch fermentation system was developed to reduce these inhibitory effects and to improve xylitol production from corn cob hemicellulose hydrolysates by C. tropicalis. RESULTS: Results of batch fermentations indicated that high xylitol production could be obtained from C. tropicalis at an initial xylose concentration of 80 g L^?1 in corn cob hydrolysate medium at an aeration rate of 0.4 vvm at the micro‐aeration stage. In the two‐stage fed‐batch fermentation process, 96.5 g L^?1 xylitol was obtained after 120 h, giving a yield of 0.83 g g^?1 and a productivity of 1.01 g L^?1 h^?1, which were 12.16% and 65.57% higher than those in a batch fermentation. CONCLUSION: High xylitol production can be achieved in a two‐stage fed‐batch fermentation process, in which the negative effects of aeration rate and inhibitory compounds on xylitol formation can be considerably reduced. Copyright © 2011 Society of Chemical Industry     

Efficient enzymatic regioselective benzoylation of 5‐fluorouridine catalysed by Novozym 435

BACKGROUND: This work focuses on developing a mild, efficient and regioselective enzymatic procedure to synthesise 5′‐O‐benzoyl‐5‐fluorouridine, a more powerful antitumour drug than 5‐fluorouridine itself, and examining the influences of several key variables on the reaction. RESULTS: The enzymatic regioselective benzoylation of 5‐fluorouridine can be successfully conducted with Novozym 435. The end‐product was confirmed to be 5′‐O‐benzoyl‐5‐fluorouridine by ¹³C nuclear magnetic resonance analysis. A co‐solvent mixture of hexane and tetrahydrofuran (THF) (50:50 v/v) was found to be the best reaction medium. The optimal initial water activity, molar ratio of vinyl benzoate to 5‐fluorouridine and reaction temperature were 0.07, 25:1 and 70 °C respectively. Under the optimised conditions the initial reaction rate, substrate conversion and regioselectivity were found to be 32.8 mmol L^?1 h^?1, 98.9% and > 99% respectively. In addition, Novozym 435 still maintained 88.2% of its original activity even after being reused for ten batches at 70 °C, indicating excellent thermal and operational stability of the enzyme in the co‐solvent mixture of hexane and THF. CONCLUSION: The results clearly show that the Novozym 435‐catalysed regioselective benzoylation of 5‐fluorouridine is a novel and facile route for efficient preparation of 5′‐O‐benzoyl‐5‐fluorouridine with potential antitumour activity. Copyright © 2008 Society of Chemical Industry     

Synthesis,Characterization and Properties of <Emphasis Type="Italic">N</Emphasis>-Alkyl-<Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis>-di(2-hydroxyethyl) Amine Oxides

N‐Dodecyl‐N,N‐di(2‐hydroxyethyl) amine oxide (C₁₂DHEAO) and N‐stearyl‐N,N‐di(2‐hydroxyethyl) amine oxide (C₁₈DHEAO) were synthesized with N‐alkyl‐diethanolamine and hydrogen peroxide. Their chemical structures were confirmed using ¹H‐NMR spectra, mass spectral fragmentation and FTIR spectroscopic analysis. It was found that C₁₂DHEAO and C₁₈DHEAO reduced the surface tension of water to a minimum value of approximately 28.75 mN m^?1 at concentration of 2.48 × 10^?3 mol L^?1 and 32.45 mN m^?1 at concentration of 5.21 × 10^?5 mol L^?1, respectively. The minimum interfacial tension (IFT_min) and the dynamic interfacial tension (DIT) of oil–water system were measured. When C₁₈DHEAO concentration was in the range of 0.1–0.5%, the IFT_min between liquid paraffin and C₁₈DHEAO solutions all reached the ultra‐low interfacial tension. Furthermore, their foam properties were investigated by Ross‐Miles method, and the height of foam of C₁₂DHEAO was 183 mm. It was also found that they showed strong emulsifying power.     

Fuel ethanol production from concentrated food waste hydrolysates in immobilized cell reactors by Saccharomyces cerevisiae H058

BACKGROUND: Continuous ethanol fermentation of concentrated food waste hydrolysates has been studied. The process was carried out in an immobilized cell reactor with beads of calcium‐alginate containing immobilized Saccharomyces cerevisiae H058 at temperature 30 °C and pH 5.0. RESULTS: The total residual sugar decreased with increase of hydraulic retention time (HRT) under various reducing sugar concentrations. Ethanol production by immobilized cells increased with increase in HRT, regardless of the substrate concentrations employed. The highest ethanol concentration of 89.28 g L^?1 was achieved at an HRT of 5.87 h and reducing sugar concentration of 200 g L^?1. At an HRT of 1.47 h, the maximum volumetric ethanol productivity of 49.88 g L^?1 h^?1 and the highest ethanol yield of 0.48 g g^?1 were achieved at reducing sugar concentration of 160 and 200 g L^?1, respectively. The difference between the fresh and the 30‐day Ca–alginate immobilized cell was also shown by scanning electronic micrographs of beads taken from their outer and inner surfaces. CONCLUSIONS: Continuous ethanol production from concentrated food waste hydrolysates using immobilized yeast cells is promising in view of the high ethanol productivity obtained at relatively high conversion and excellent reactor stability. Copyright © 2011 Society of Chemical Industry