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1.
Biotransformation of benzaldehyde to L-phenylacetylcarbinol (L-PAC) as a key intermediate for L-ephedrine has been evaluated using immobilized pyruvate decarboxylase (PDC) from Candida utilis. PDC immobilized in spherical polyacrylamide beads was found to have a longer half-life compared with free enzyme. In a batch process, the immobilized PDC generally produced lower L-PAC than free enzyme at the same concentrations of substrates due to increased by-products acetaldehyde and acetoin and reduced benzaldehyde uptake. With immobilized PDC, L-PAC formation occurred at higher benzaldehyde concentrations (up to 300 mM) with the highest L-PAC concentration being 181 mM (27.1 g/L). For a continuous process, when 50 mM benzaldehyde and 100 mM sodium pyruvate were fed into a packed-bed reactor at 4 degrees C and pH 6.5, a productivity of 3.7 mM/h (0.56 g/L . h) L-PAC was obtained at an average concentration of 30 mM (4.5 g/L). The half-life of immobilized PDC reactor was 32 days. (c) 1996 John Wiley & Sons, Inc.  相似文献   

2.
Biotransformation of benzaldehyde to L-phenylacetylcarbinol (L-PAC) as a key intermediate for L-ephedrine synthesis has been evaluated using pyruvate decarboxylase (PDC) partially purified from Candida utilis. PDC activity was enhanced by controlled fermentative metabolism and pulse feeding of glucose prior to the enzyme purification. With partially purified PDC, several enzymatic reactions occurred simultaneously and gave rise to by-products (acetaldehyde and acetoin) as well as L-PAC production. Optimal reaction conditions were determined for temperature, pH, addition of ethanol, PDC activity, benzaldehyde, and pyruvate:benzaldehyde ratio to maximize L-PAC, and minimize by-products. The highest L-PAC concentration of 28.6 g/L (190.6 mM) was achieved at 7 U/mL PDC activity and 200 mM benzaldehyde with 2.0 molar ratio of pyruvate to benzaldehyde in 40 mM potassium phosphate buffer (pH 7.0) containing 2.0 M ethanol at 4 degrees C. (c) 1996 John Wiley & Sons, Inc.  相似文献   

3.
The effect of process parameters on the biotransformation of benzaldehyde to L-phenylacetylcarbinol (L-PAC) using a yeast isolate identified as Torulaspora delbrueckii was studied. The maximum yield of L-PAC obtained was (331 mg) per 100 ml biotransformation medium (glucose 3%, peptone 0.6% and at pH 4.5) from 600 mg of benzaldehyde with 8 h of reaction at 30 ± 2 °C. Growing the organism in presence of 3% glucose reduced the biotransformation time to 120 min. Addition of 0.6% acetaldehyde (30–35%) lead to an increase in L-PAC yield to 450 mg%. Semi-continuous feeding of benzaldehyde (200 mg) and acetaldehyde (200 l) four times at 30 min intervals could produce 683 mg of L-PAC/100 ml biotransformation medium. Chiral HPLC analysis of purified L-PAC and PAC-diol showed 99% enantiomeric purity. The cell mass was found to be reusable for biotransformation up to nine times when benzaldehyde and acetaldehyde levels were maintained at (350 mg and 350 l)–(400 mg and 400 l). At concentrations from 450 mg and 450 l to 600 mg and 600 l, however the cell mass could give efficient biotransformation only during one use.  相似文献   

4.
To provide further understanding of the biotransformation of benzaldehyde to L-phenylacetyl carbinol (L-PAC), an intermediate in L-ephedrine production, a kinetic model has been developed for the deactivation of pyruvate decarboxylase (PDC) by benzaldehyde. The model confirms that deactivation is first order with respect to benzaldehyde concentration and exhibits a square root dependency on time. The model covers the range of benzaldehyde concentrations 100–300 mM, as it has been shown previously that 200 mM benzaldehyde can produce L-PAC concentrations up to 190 mM (28.6 g/L) using partially purified PDC from Candida utilis.  相似文献   

5.
Immobilization of Saccharomyces cerevisiae ATCC 834 within alginate beads enhances microbiological conversion of benzaldehyde to L-phenylacetyl carbinol (L-PAC), a precursor employed for synthesis of L-ephedrine. Yields of 90% L-PAC on benzaldehyde (initially 0.6% in medium) were obtained with immobilized cells, in contrast to about 10% with free cells which tend to form pellets in the presence of benzaldehyde. The predominant favorable action of immobilization appears to be a reduction in the toxic or inhibitory effects of benzaldehyde. With an initial benzaldehyde concentration of about 0.6% in the medium the optimum cell mass concentration was observed to be about 28 g cell mass (immobilized) per liter of medium.  相似文献   

6.
The cyclic, semicontinuous production of L-phenylacetyl carbinol (L-PAC) from a benzaldehyde substrate by Saccharomyces cerevisiae ATCC 834 immobilized in calcium alginate beads was substantially enhanced to about 4.5 g/L in a second cycle by reactivation in fresh medium for 24 h, following an earlier 24-h period of production from substrate. Intermittent feeding of benzaldehyde was employed (four doses in 3 h). In subsequent similar cycles, however, the production returned to that produced in the first cycle, viz. L-PAC concentration of 2-3 g/L in the medium. Production of L-PAC was also increased by adaptation of the cells over 200 h of exposure to the benzaldehyde substrate (compared to wild-type cells) and by continuous (as compared to intermittent) feeding of the substrate. A liter as great as 10 g/L was obtained with wild-type cells by continuous feeding of benzaldehyde over 6 h. Immobilization not only protected the cells from toxic effects of substrate but also permitted them to be used during 7 cycles of semicontinuous operation over more than 200 h.  相似文献   

7.
L-Phenylacetylcarbinol (L-PAC): biosynthesis and industrial applications   总被引:1,自引:0,他引:1  
L-Phenylacetylcarbinol (L-PAC), an important drug intermediate, can be produced by biotransformation of benzaldehyde, mainly by yeast cultures but also by Zymomonas mobilis. The biotransformation by free cells, immobilized cells, mutant organisms, purified pyruvate decarboxylase as well as the use of bioreactors, the downstream processing of L-PAC and the industrial applications have been reviewed. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

8.
The gas-liquid mass transfer coefficient K(L)a in the fermenter is a strong function of mode of energy dissipation and physico-chemical properties of the liquid media. A combination of disc turbine (DT) and pitched blade turbine down flow (PTD) impellers has been tested in laboratory bioreactor for gas hold-up and gas-liquid mass transfer performance for the growth and biotransformation medium for an yeast isolate VS1 capable of biotransforming benzaldehyde to L-phenyl acetyl carbinol (L-PAC) and compared with those in water.Correlations have been developed for the prediction of the fractional gas hold-up and gas-liquid mass transfer coefficient for the above media. The mass transfer coefficient and respiration rate have been determined in the shake flask for the growth as well as for biotransformation medium. These results, then have been used to optimize the operating parameters (impeller speed and aeration) for growth and biotransformation in a laboratory bioreactor. The comparison of cell mass production and L-PAC production in the bioreactor has been done with that obtained in shake flask studies.  相似文献   

9.
The rate and extent of microbial transformation of higher concentrations of benzaldehyde substrate to L-phenylacetyl carbinol (L-PAC) by immobilized cells of Saccharomyces cerevisiae ATCC 834 was markedly stimulated by addition of different concentrations of beta-cyclodextrin (BCD) to the fermentation medium. With 0.5, 1.0, and 1.5% BCD in the fermentation medium and cumulative doses of benzaldehyde of 12 and 14 g/L, significantly higher yields of L-PAC were obtained, about one- to twofold that of the yields of the control experiments. The favorable effects of BCD were evident in spite of its presence in stoichiometric concentrations significantly lower than those of benzaldehyde. The presence of BCD also appeared to stimulate microbial growth slightly. Enhanced cellular activity was reflected by faster D-glucose consumption and faster benzaldehyde utilization in the presence of BCD.  相似文献   

10.
Summary L-PAC production by a yeast strain harvested from glucose limited continuous cultures grown at different dilution rates has been studied. The specific transformation rate is higher for the cells which have been grown at higher dilution rates.C.D.R.I. Communication No. 4296  相似文献   

11.
Conversion of benzaldehyde to L-phenylacetyl carbinol (L-PAC) was achieved with immobilized, growing cells of Saccharomyces cerevisiae in different reactors. Product formation increased (31%) with the subsequent initial reuses of the entrapped cells. Biomass production and PAC formation depleted (40 and 57%, respectively) after 4-5 continuous growth and biotransformation cycles. With the regeneration of the biocatalysts, catalytic activity of the cells was resumed. The highest yields were in a stirred tank reactor (29 g PAC) from 77 g benzeldehyde with 14 repeated uses of entrapped cells.  相似文献   

12.
PurposeThe purpose of this study was to; (1) investigate employing a novel position-sensitive mega-size polycarbonate (MSPC) dosimeter for photoneutron (PN) depth, profile and dose equivalent distributions studies in a multilayer polyethylene phantom in a Siemens ONCOR accelerator, and (2) develop depth dose equivalent distribution matrix data at different depths and positions of the phantom for patient PN dose equivalent determination and in particular for PN secondary cancer risk estimation.MethodsPosition-sensitive MSPC dosimeters were successfully exposed at 9 different depths of the phantom in a 10 × 10 cm2 X-ray field. The dosimeters were processed in mega-size electrochemical chambers at optimum conditions. Each MSPC dosimeter was placed at a known phantom depth for PN depth dose equivalents and profiles on transverse, longitudinal and diagonal axes and isodose equivalent distribution studies in and out of the X-ray beam.ResultsPN dose equivalent distributions at any depth showed the highest value at the beam central axis and decreases as the distance increases. PN dose equivalent at any position studied in the axes has a maximum value on the phantom surface which decreases as depth increases due to flux reduction by multi-elastic scattering interactions.ConclusionsExtensive PN dose equivalent matrix data at different depths and positions in the phantom were determined. The position-sensitive MSPC dosimeters proved to be highly efficient for PN depth, profile and isodose equivalent distribution studies. The extensive data obtained highly assists for determining PN dose equivalent of a patient undergoing high-energy X-ray therapy and for PN secondary cancer risk estimation.  相似文献   

13.
The mechanism of embryonic polarity establishment in mammals has long been controversial. Whereas some claim prepatterning in the egg, we recently presented evidence that mouse embryonic polarity is not established until blastocyst and proposed the mechanical constraint model. Here we apply computer simulation to clarify the minimal cellular properties required for this morphology. The simulation is based on three assumptions: (1) behavior of cell aggregates is simulated by a 3D vertex dynamics model; (2) all cells have equivalent mechanical properties; (3) an inner cavity with equivalent surface properties is gradually enlarged. However, an initial attempt reveals a requirement for an additional assumption: (4) the surface of the cavity is firmer than intercellular surfaces, suggesting the presence of a basement membrane lining the blastocyst cavity, which is indeed confirmed by published data. The simulation thus successfully produces a structure recapitulating the mouse blastocyst. The axis of the blastocyst, however, remains variable, leading us to an additional assumption: (5) the aggregate is enclosed by a capsule, equivalent to the zona pellucida in vivo. Whereas a spherical capsule does not stabilize the blastocyst axis, an ellipsoidal capsule eventually orients the axis in accordance with its longest diameter. These predictions are experimentally verified by time-lapse recordings of mouse embryos. During simulation, equivalent cells form two distinct populations composed of smaller inner cells and larger outer cells. These results reveal a unique feature of early mammalian development: an asymmetry may emerge autonomously in an equivalent population with no need for a priori intrinsic differences.  相似文献   

14.
The study aimed to evaluate extraction efficiency, detection and quantification of phytochemicals, minerals and antioxidative capacity of different parts of Salacia chinensis L. Continuous shaking extraction, steam bath assisted extraction, ultrasonic extraction and microwave assisted extraction with varied time intervals were employed for extraction of phenolics, flavonoids, and antioxidants. Preliminary screening revealed the presence of wide array of metabolites along with carbohydrates and starch. Steam bath assisted extraction for 10 min exposure was found most suitable for extraction phenolics (46.02 ± 2.30 mg of gallic acid equivalent per gram of dry weight and 48.57 ± 2.42 mg of tannic acid equivalent per gram of dry weight) and flavonoids (35.26 ± 1.61 mg of quercetin equivalent per gram of dry weight and 51.60 ± 2.58 mg of ellagic acid equivalent per gram of dry weight). In support, reverse phase-high performance liquid chromatography- diode array detector confirmed the presence of seven pharmaceutically important phenolic acids. Antioxidant capacity was measured by 1, 1- diphenyl-1-picryl hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), 2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) scavenging (ABTS) and N, N-dimethyl-p-phenylenediamine (DMPD) assays and represented as trolox equivalent antioxidant capacity (TEAC) and ascorbic acid equivalent antioxidant capacity (AEAC). Antioxidant capacity ranged from 121.02 ± 6.05 to 1567.28 ± 78.36 µM trolox equivalent antioxidant capacity and 56.62 ± 2.83 to 972.48 ± 48.62 µM ascorbic acid equivalent antioxidant capacity. Roots showed higher yields of illustrated biochemical parameters, however fresh fruit pulp was found a chief source of minerals. Gas chromatography-mass spectroscopic analysis revealed the presence of a vast array of phytoconstituents associated with different plant parts. The present study revealed the amounts of minerals and diverse phytoconstituents in various parts of S. chinensis and confirmed its medicinal and nutritional implications.  相似文献   

15.
The problem of obtaining Helmholtz equivalents for nonlinear resistive one-ports is considered. Two fundamentally different classes of equivalent are described, one local and the other global. For each, necessary and sufficient conditions are derived for the existence and uniqueness of either the Thévenin equivalent or the Norton equivalent or both. These concepts are illustrated (i) by proving that a cell whose channels and pumps are monotone in the membrane potential will, in the absence of net state changes in these ionophores, possess a unique stable resting potential and (ii) by demonstrating that it is in principle impossible to assign unique equivalent circuits to such ionophores.  相似文献   

16.
Taraxacum kok‐saghyz (TKS) is a dandelion species native to Kazakhstan, Uzbekistan and north‐west China, considered as a promising alternative source of natural rubber from its roots. The aim of this study was to investigate the possible exploitation of TKS leaves, a rubber byproduct, as a source of phenolic compounds with antioxidant properties for potential applications in forage, nutraceutical and pharmacological fields. Two accessions (TKS016, TKS018) grown under Mediterranean conditions of Sardinia were evaluated at vegetative and flowering stages. The leaves of TKS018 had the highest antioxidant capacity (19.6 mmol trolox equivalent antioxidant capacity 100 g?1), total phenolic (106.4 g gallic acid equivalent kg?1), tannic phenolics (58.5 g gallic acid equivalent kg?1) and total flavonoid contents (22.9 g catechin equivalent kg?1). At both phenological stages, TKS016 showed significantly lower values than TKS018 in 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), total phenolic and tannic phenolics. Six individual molecules were identified, namely chlorogenic, cryptochlorogenic, caffeic, sinapic, chicoric and 3,4‐dimethoxycinnamic acids. Chicoric (8.53–10.68 g kg?1 DW) and chlorogenic acids (4.18–7.04 g kg?1 DW) were the most abundant. TKS leaves represent a valuable source of chicoric acid with potential application as antioxidant to be used as herbal medicine and nutrition for production of healthy food/feed.  相似文献   

17.
Tetra-2-pyridyl-1,4-diazine (TPD) reacts with one equivalent of Ru(tpy)Cl3 to provide (tpy)Ru(TPD)2+ which when treated with a second equivalent of Ru(tpy)Cl3 under more forcing conditions provides the TPD bridged binuclear complex, (tpy)Ru(TPD)Ru(tpy)4+. The structure of both complexes, particularly with regard to planarity of the pendant pyridine rings is analyzed by high field NMR. The electronic spectrum of the binuclear complex shows pronounced bathochromic shift, while the redox potentials for (tpy)Ru(TPD)2+. indicate a diminished HOMO-LUMO energy gap.  相似文献   

18.
European field experiments have demonstrated Miscanthus can produce some of the highest energy yields per hectare of all potential energy crops. Previous modelling studies using MISCANMOD have calculated the potential energy yield for the EU27 from mean historical climate data (1960–1990). In this paper, we have built on the previous studies by further developing a new Miscanthus crop growth model MISCANFOR in order to analyse (i) interannual variation in yields for past and future climates, (ii) genotype-specific parameters on yield in Europe. Under recent climatic conditions (1960–1990) we show that 10% of arable land could produce 1709 PJ and mitigate 30 Tg of carbon dioxide-carbon (CO2-C) equivalent greenhouse gasses (GHGs) compared with EU27 primary energy consumption of 65 598 PJ, emitting 1048 Tg of CO2-C equivalent GHGs in 2005. If we continue to use the clone Miscanthus × giganteus , MISCANFOR shows that, as climate change reduces in-season water availability, energy production and carbon mitigation could fall 80% by 2080 for the Intergovernmental Panel on Climate Change A2 scenario. However, because Miscanthus is found in a huge range of climates in Asia, we propose that new hybrids will incorporate genes conferring superior drought and frost tolerance. Using parameters from characterized germplasm, we calculate energy production could increase from present levels by 88% (to 2360 PJ) and mitigate 42 Tg of CO2-C equivalent using 10% arable land for the 2080 mid-range A2 scenario. This is equivalent to 3.6% of 2005 EU27 primary energy consumption and 4.0% of total CO2 equivalent C GHG emissions.  相似文献   

19.
In this paper, we consider the concepts of evolutionarily stable strategy (ESS), neighborhood invader strategy (NIS) and global invader strategy (GIS) in single species with frequency-dependent interactions. We find some general relationships among the three concepts in matrix games. The main conclusion is that ESS and NIS are equivalent to each other and are both equivalent to local superiority; a strategy with global superiority must be a GIS; a GIS may not be equivalent to its global superiority in games with more than two players; and in any two-player matrix game a GIS is just equivalent to its global superiority. In two-player games, globally asymptotic stability in the replicator dynamics has also been shown. Equivalent conditions for the three concepts stated by payoff comparisons are given and are applied to examples involved.  相似文献   

20.
The purpose of this study was to explore the effect of the second bend of the maze, on the neutron dose equivalent, in the 15 MV linear accelerator vault, with two bend maze. These two bends of the maze were covered by 32 points where the neutron dose equivalent was measured. There is one available method for estimation of the neutron dose equivalent at the entrance door of the two bend maze which was tested using the results of the measurements. The results of this study show that the neutron equivalent dose at the door of the two bend maze was reduced almost three orders of magnitude. The measured TVD in the first bend (closer to the inner maze entrance) is about 5 m. The measured TVD result is close to the TVD values usually used in the proposed models for estimation of neutron dose equivalent at the entrance door of the single bend maze. The results also determined that the TVD in the second bend (next to the maze entrance door) is significantly lower than the TVD values found in the first maze bend.  相似文献   

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