郁李仁(R)-醇腈酶高效催化乙酰基三甲基硅烷与丙酮氰醇不对称转氰

Highly efficient asymmetric transcyanation of acetyltrimethylsilane with acetone cyanohydrin in an aqueous/organic biphasic system catalyzed with(R)-oxynitrilase from defatted Prunus Japonica seed meal for the preparation of optically active(R)-2-trimethylsilyl-2-hydroxyl-propionitrile was successfully carried out for the first time.For better understanding of the reaction,various influential variables were examined with respect to the initial reaction rate,the substrate conversion and the product enantiomeric excess(e.e.).Diisopropyl ether was found to be the best organic phase for this reaction among all the organic solvents tested.The optimal concentrations of Prunus Japonica seed meal powder,acetyltrimethylsilane and acetone cyanohydrin,volume ratio of aqueous phase to organic phase,buffer pH value and the reaction temperature were 34. 5 g.L~(-1) and 14 mmol·L~(-1) ,28 mmol.L~(-1) , 13%(by volume),5. 0 and 30℃,respectively,while the initial reaction rate,the substrate conversion and the product enantiomeric excess were 1. 34 mmol.L~(-1) .h~(-1) ,99. 0% and 99. 0%,respectively.The comparative study demonstrated that silicon atom in substrate showed great effect on the reaction and acetyltrimethylsilane was a much better substrate for(R)-hydroxynitrile lyase from Prunus Japonica seed than its carbon analogue 3,3-dimethyl-2-butanone.     

水/有机溶剂双相中固定化啤酒酵母细胞催化三甲基硅乙酮不对称还原

Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied,The effects of shake speed,hydrophobictiy of organic solvent ,volume ratio of water phase to organic phase,pH value of aqueous phase and reaction temperature on the initial reaction rate,maximum yield and enantiomeric excess(ee) of the product were systematically explored,All the above-mentioned factors had significant infuence on the reaction.n-Hexane was found to be the best organic solvent for the reaction.The optimum shake speed,volume ratio of water phase to organic phase,pH value and reaction temperature were 150 r.min^-1,1/2,8 and 30℃ respectively,under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%,which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase ,To our best knowledge,this is the most satisfactory result obtained.     

Asymmetric Synthesis of (-)-1-Trimethylsilyl-ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Diphasic System

Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied. The effects of shake speed, hydrophobicity of organic solvent, volume ratio of water phase to organic phase, pH value of aqueous phase and reaction temperature on the initial reaction rate, maximum yield and enantiomeric excess (ee) of the product were systematically explored. All the above-mentioned factors had significant influence on the reaction. n-Hexane was found to be the best organic solvent for the reaction. The optimum shake speed, volume ratio of water phase to organic phase, pH value and reaction temperature were 150 r.min-1, 1/2, 8 and 30 ℃ respectively, under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%, which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase. To our best knowledge, this is the most satisfactory result obtained.     

有机溶剂中树脂催化丙酮氰醇分解反应的动力学模型

Decomposition of acetone cyanohydrin is the first-step reaction for preparing (S)-α-cyano-3-phenoxybenzyl alcohol (CPBA) by the one-pot method in organic media. Considering the compatibility of bio-catalysts with chemical catalysts and the successive operation in the bioreactor, anion exchange resin (D301) was used as catalyst for this reaction. External diffusion limitation was excluded by raising rotational speed to higher than 190r·min-1 in both solvents. Internal diffusion limitation was verified to be insignificant in this reaction system. The effect of acetone cyanohydrin concentration on the reaction was also investigated. An intrinsic kinetic model was proposed when the mass transfer limitation was excluded, and the average deviation of the model is 10.5%.     

有机介质中脂肪酶促动力学拆发外消旋1-三甲基硅乙醇

The enantioselective esterification of racemic 1-trimethylsilylethanol with acids catalyzed by lipase in organic solvent was successfully performed. The influence of some factors on the reaction was investigated. Among the four lipases explored, Candlda rugosa lipase (CRL) showed the highest activity and enantioselectivity. Octanoic acid was the best acyl donor among the eleven acids studied and n-hexane was the most suitable medium for the reaction. The optimum shaking rate and temperature were found to be 150 r-rain-i and 20~(3 to 30~C, respectively.The enantiomeric excess of the remaining (S)-(-)-1-trimethylsilylethanol was 93% when substrate conversion was 53% upon incubation of the reaction mixture at 30~C, 150 r-rain-i for 12 h.     

Kinetics of Asymmetric Reduction of Phenylglyoxylic Acid to R-(－)-Mandelic Acid by Saccharomyces Cerevisiae FD11b

The kinetics of asymmetric production of R-(-)-mandelic acid (R-MA) from phenylglyoxylic acid (PGA)catalyzed by Saccharomyces cerevisiae sp. strain FD11b was studied by fed-batch cultures. The concentrations of glucose and PGA were controlled respectively with a dual feeding system. When the electron donor glucose was supplied at the rate of 0.0833mmol·gdw-1·h-1, the specific production rate (qp) and the enantiomeric excess of R-MAreached the maximum 0.353mmol·gdw-1·h-1 and 97.1％, respectively. The apparent reduction activity of yeast FD11b was obviously affected by both substrate PGA and product MA. The qp value reached the maximum 0.36-0.38mmol·gdw-1·h-1 when the PGA concentration was controlled between 25 and 35mmol·L-1. The obvious substrate inhibition of bioconversion was observed at the PGA concentrations higher than 40mmol· L-1. The accumulation of product MA also caused a severe feed-back inhibition for its production when the product concentration was above 60mmol· L-1. The kinetic model with the inhibition effect of both substrate and product was simulated by a computer-based least-square arithmatic. The established kinetic model was in good agreement with the experimental data.     

阿魏酸对蘑菇酪氨酸酶的抑制效应

The inhibitory effect of ferulic acid on the diphenolase activity of mushroom tyrosinase and the kinetic behavior were studied with L-3,4-dihydroxyphenylalanine （L-DOPA） as substrate. The inhibitor concentration leading to 50% relative activity lost （IC50） was estimated to be 0.15 mmol·L^-1. The inhibition mechanism obtained from Lineweaver-Burk plots shows that ferulic acid is a competitive inhibitor and the inhibition of tyrosinase by ferulic acid is a reversible reaction. The equilibrium constant for ferulic acid binding with the tyrosinase was determined to be 0.25 mmol·L^-1 for diphenolase. Keywords tyrosinase, ferulic acid, kinetics, inhibition, L-DOPA, diphenolase     

表面活性剂包衣Candida rugosa脂肪酶在无溶剂下油水两相体系中催化橄榄油水解

The surfactant-coated Candida rugosa lipase was used as catalyst for hydrolysis of olive oil in two-phase system consisting of olive oil and phosphate buffer without organic solvent. For both the coated and native lipases,the optimal buffer/oil volume ratio of 1.0, aqueous pH 6.8 and reaction temperature 30℃ were determined. The maximum activity of the coated lipase was ca 1.3 times than that of the native lipase. The half-life of the coated lipase in olive oil and the native lipase in phosphate buffer was ca 9 h and 12 h, and the final residual activity was 27% and 20% of their initial values, respectively. The final substrate conversion by the coated lipase was ca 20% higher than that of the native lipase.     

Direct extraction of Mo(Ⅵ) from sulfate solution by synergistic extractants in the rotation column

Direct extraction of molybdenum from sulfate solution with synergistic extractants(mixture of D_2EHPA and TBP)was studied in the rotation column. The influence of extractant concentration and initial pH of aqueous phase was studied in the bench scale experiments. The outcomes demonstrated that the synergistic solvent extraction enhances the constancy of the extracted complexes for transfer into the organic phase. In the continuous experiments, the effect of different operating parameters such as speed of agitation, inlet solvent flow rate and inlet aqueous flow rate on the holdup, mean drop size, drop size distribution, slip and characteristic velocities and extraction percentage were examined. Modified correlations were proposed for prediction of hydrodynamic parameters with consideration of reaction extraction condition in the rotation column. Furthermore, these correlations were compared with the experimental data. According to the results, the direct extraction of Mo(Ⅵ) from aqueous solution and sulfuric media with extraction efficiency of 90.4% was obtained at higher rotor speed(240 r·min~(–1) rpm) in this column.     

Enantioselective esterification of (R,S)-2-(4-methylphenyl) propionic acid via Novozym 435: Optimization and application

This paper reports on the resolution of(R,S)-2-(4-methylphenyl) propionic acid(MPPA) enantiomers by enzymatic esterification in organic solvent. Novozym 435(CALB) has the best catalytic performance compared with other lipases. Of the alcohols screened, n-hexanol is the best acyl acceptor and gives the highest enzyme activity and enantioselectivity in n-hexane. Response surface methodology(RSM) was used to evaluate the influence of the factors, such as temperature, enzyme amount, substrate concentration and reaction time on the substrate conversion(c) and enantiomeric excess(ee). The correlation coefficient R~2 for enantiomeric excess and the conversion are 0.9827 and 0.9910, respectively, indicating that can accurately predict the experimental results. By simulation and optimization, the optimal conditions were obtained, involving 600 mmol·L~(-1) MPPA concentration(0.60 mmol), 850 mmol·L~(-1) hexanol concentration(0.85 mmol), 58 mg enzyme amount, 75 ℃ temperature and 4.5 h reaction time, respectively. Under the optimized conditions, the experimental values of conversion and enantiomeric excess were 89.34% and 97.84%, respectively, which are in good agreement with the model predictions.     

乳状液膜包酶制备6-APA的研究

Production of 6-aminopenicillanic acid (6-APA) by hydrolysis using penicillin acylase (PA) was studied as a model of an enzymatic emulsion liquid membrane (ELM) process. The loss of PA activity was examined for various membrane compositions (organic solvent, surfactant, carrier). The effects of some experimental variables on the stability of emulsion were investigated. It was found that the choice of organic solvent greatly affected tilestability of the emulsion. Increasing the concentration of the carrier in the membrane phase increases the transfer rate of substrate and products but also has a destabilizing effect on the emulsion. The recovery of 6-APA obtained by a di-carrier system (N263-N1923) was much higher than those when either of the di-carriers was used separately.The whole process was controlled both by the enzymatic reaction rate and by the transfer rate of the substrate and the products, however, the ratio of them could be changed by varying the composition of the system. For an optimum condition, it was obtained that the recovery ratio of 6-APA was over 80% and the conversion of benzyl penicillin (PG) was up to 90% in the external phase after 30 minutes. Meanwhile, the breakage percentage of the emulsion was less than 2%.     

丙酸盐影响利迪链霉菌AS 4.2501生产利迪链菌素及其碳流分布

To achieve higher antibiotic streptolydigin productivity through metabolic regulation, propionate was fed during the fermentation of Streptomyces lydicus AS 4.2501. The effects of propionate feeding on streptolydigin production and intracellular fluxes were investigated. The highest streptolydigin production yield of 95.10mg·L^-1 was obtained when 2mmol·L^-1 of sodium propionate was added at 60h of cultivation into shake-flask culture. This yield is 23.06% higher when compared to that of a batch culture without propionate supplementation. It was also found that when propionate was added, much more organic acids were excreted. Metabolic flux analysis was performed and it demonstrated that the carbon fluxes of the pentose phosphate pathway and the anaplerotic reaction were significantly increased after propionate feeding. The carbon flux from pyruvate to acetyl-CoA was determined to be 24.7, which was 12.27% higher than that in the batch culture. This study indicated that the glucose-6-phosphate and pyruvate nodes were potential bottlenecks for increasing streptolydigin productivity. Potential targets and strategies that could be manipulated through genetic and process engineering to increase the production of streptolydigin were also suggested.     

重组大肠杆菌分批-补料高密度发酵生产类人胶原蛋白的动力学

The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.     

伴随传质的液液体系微分散流型图的实验研究(英文)

This paper presents the experimental results of liquid-liquid microflows in a coaxial microfluidic device with mass transfer.Three working systems were n-butanol + phosphoric acid(PA) + water,methyl isobutyl ketone(MIBK) + PA + water,30% kerosene in tri-n-butylphosphate(TBP) + PA + water.The direction and intensity of mass transfer were adjusted by adding PA in one of two phases mutual saturated in advance.When PA transferred from the organic phase to the aqueous phase,tiny aqueous droplets may generate inside the organic phase by mass transfer inducement to form a new W/O/W flow pattern directly on some special cases.Once the PA concentration was very high,violent Marangoni effect could be observed to throw part of organic phase out of droplets as tail.The interphase transfer of PA could expand the jetting flow region,in particular for systems with low or medium inter-facial tension and when the mass transfer direction was from the aqueous phase to the organic phase.     

以L-谷氨酰胺-铜（Ⅱ）配合物为供体酶法制备茶氨酸

A novel method for enzymatic synthesis of L-theanine using copper（Ⅱ）-L-glutamine［Cu（Gln）₂］as donor substrate was proposed.The structure of Cu（Gln）₂ was identified by infrared spectrum analysis and its stability was also investigated under the reaction conditions.The enzymatic synthesis of L-theanine catalyzed by γ-glutamyltranspeptidase was carried out by using Cu（Gln）₂ and L-glutamine as donor substrate respectively,and the product yield and conversion rate of donor substrate were compared under the conditions of different ratios of donor and acceptor.The results showed that the transpeptidation reaction could be effectively enhanced by using Cu（Gln）₂ as donor substrate.The conversion rate of donor would increase by 51.5%,44.9% and 27.1% under different reaction conditions,compared to that using L-Gln as donor substrate.When the molar ratio of donor to acceptor was 6∶100,a higher donor conversion of 71.3% was obtained.     

RADICAL DESORPTION IN EMULSION POLYMERIZATION OF VINYL ACETATE

In the present paper a model equation for calculating the radical desorption rate constant,k_0,in the emulsion polymerization of vinyl acetate was suggested and the various parameters forevaluating k_0 were determined.Effects of reaction temperature,emulsifier concentration,initiator con-centration,monomer conversion and phase ratio on k_0 were studied.It indicates that the desorptionof radicals from latex particles to aqueous phase must be taken into account in the modelling ofemulsion polymerization for the monomers with higher hydrophilicity such as vinyl acetate.     

Selective oxidation of cyclopentene with H_2O_2 by using H_3PW_(12)O_(40) and TBAB as a phase transfer catalyst

The selective oxidation of cyclopentene by aqueous H_2O_2 using H_3PW_(12)O_(40) and tetrabutyl ammonium bromide(TBAB) as a phase transfer catalyst has been investigated. The results show that the presence of TBAB significantly improved the oxidation selectivity of cyclopentene. The effects of the reaction conditions on the conversion of cyclopentene were investigated in detail. The optimal reaction conditions are as follows: the H_3PW_(12)O_(40) to TBAB molar ratio, 1:1–1:3; H_3PW_(12)O_(40) to cyclopentene molar ratio,0.54:100–0.64:100; and molar ratio of H_2O_2 to cyclopentene, 1.6:1. The conversion reached to 59.8% in 4h at 35.0 °C, while the selectivity of glutaraldehyde was 38.0% and the selectivity of 1,2-cyclopentanediol was 55.6%. In addition, a route for oxidation of cyclopentene by aqueous H_2O_2 using a heteropoly acid and quaternary ammonium salt as a phase transfer catalyst was proposed.     

有机溶剂原位萃取法提高悬浮培养中国红豆杉细胞紫杉醇产量

The production of paclitaxel from suspension culture of Taxus chinensis var,mairei was improved by in situ extraction with organic solvents to avoid feedback repression and product degradation.Oleic acid and dibutyl phthalate were proved to be suitable solvents .The optimal volumetric percentage of organic solvents in the culture medium was found to be around 8%,and the favorable time for their introduction was at the exponential phase of cell growth,Paclitaxel production with the in situ extraction was ca 3-fold of that without extraction.     

两步酶法合成阿糖鸟苷

The chemical synthesis of Guanine arabinoside （ara-G） is extremely complex, time-consuming, and seriously polluted. A two-step enzymatic synthesis process was developed to acquire ara-G easily. 2,6-Diaminopurine arabinoside （ara-DA） was first synthesized with purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase produced by Enterobacter aerogenes DGW-07. The conversion yield of ara-DA could reach above 90% when the reaction liquid contained 30 mmol·L^-1 uracil arabinoside as arabinose donor, 10 mmol·L^- 1 2,6-diaminopurine as arabinose acceptor in pH 7.0 20 mmol·L^-1 phosphate buffer, and reacted at 60℃ for 48h. Then, ara-DA was effectively transformed into ara-G with adenylate deaminase produced by Aspergillus oryzae DAW-01. The total process had no complex separation and purification.     

热带假丝酵母不对称还原3,5-双三氟甲基苯乙酮制备手性醇(英文)

(S)-3,5-bistrifluoromethylphenyl ethanol is a key chiral intermediate for the synthesis of NK-1 receptor antagonists. Enantioselective synthesis of (S)-3,5-bistrifluoromethylphenyl ethanol was successfully performed in high enantiomeric excess (e.e.) through asymmetric reduction of 3,5-bis(trifluoromethyl) acetophenone catalyzed by Candida tropicalis 104 cells. The influence of some key reaction parameters such as substrate concentration, co-substrate and its concentration, biomass and reaction time was examined, respectively. The results showed that these factors obviously influence the yield, but the optical purity of the prepared product remains intact. The opti-mum conditions for the preparation of (S)-3,5-bistrifluoromethylphenyl ethanol were found to be as follows: sub-strate concentration 50 mmol?L?1; 50 g·L-1 of maltose as co-substrate; wet cell concentration 300 g·L-1; reaction for 30 h. Under above optimal conditions, the maximum yield for (S)-3,5-bistrifluoromethylphenyl ethanol reached 70.3% with 100% of product e.e.