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1.
Experimental and theoretical studies are conducted to understand the dynamic behavior of a continuous-flow fixed-bed reactor in which an esterification is catalyzed by an immobilized enzyme in an organic solvent medium. The experimental system consists of a commercial immobilized lipase preparation known as Lipozyme as the biocatalyst, with propionic acid and isoamyl alcohol (dissolved in hexane) as the reaction substrates. A complex dynamic behavior is observed experimentally as a result of the simultaneous occurrence of reaction and adsorption phenomena. Both propionic acid and water are adsorbed by the biocatalyst resulting in lower reaction rates. In addition, an excessive accumulation of water in the reactor leads to a rapid irreversible inactivation of the enzyme. A model based on previously-obtained adsorption isotherms and kinetic expressions, as well as on adsorption rate measurements obtained in this work, is used to predict the concentration and thermodynamic activity of water along the reactor length. The model successfully predicts the dynamic behavior of the reactor and shows that a maximum thermodynamic activity of water occurs at a point at some distance from the reactor entrance. A cation exchange resin in sodium form, packed in the reactor as a selective water adsorbent together with the catalyst particles, is shown to be an effective means for preventing an excessive accumulation of water formed in the reaction. Its use results in longer cycle times and greater productivity. As predicted by the model, the experimental results show that the water adsorbed on the catalyst and on the ion exchange resin can be removed with isoamyl alcohol with no apparent loss in enzyme activity. 相似文献
2.
Reducing the influence of an undesired product in an enzymatic reaction could have a significant impact on the productivity of such systems. Here, we focus on the removal of water formed during an enzymatic esterification in a batch reactor. A commercial immobilized lipase preparation, known as Lipozyme, is used as the biocatalyst and propionic acid and isoamyl alcohol dissolved in hexane are the substrates. In this system, the water formed will partition between the catalyst and the medium. As the more polar reactants are converted into the less polar ester product, the water is partitioned more towards the biocatalyst and the accumulation of water eventually causes lower reaction rates. Addition of a strong-acid cation exchange resin in sodium form is found to control the water accumulation on the biocatalyst without stripping the essential water needed for the enzyme to function and substantial improvements in conversion are achieved. A mathematical model is developed to describe the batch reaction behavior with and without added absorbent, which successfully predicts the behavior of water and its effects. 相似文献
3.
Regioselective enzymatic diol esterification in batch and fixed-Bed adsorptive reactors: experiments and modeling 总被引:1,自引:0,他引:1
The dynamic behavior of batch and fixed-bed adsorptive reactors is studied for the enzyme-catalyzed regioselective esterification of propionic acid and 2-ethyl-1,3-hexanediol in hexane. The reaction is equilibrium-limited with an apparent equilibrium constant of 0.6 +/- 0.1 at 22 degrees C. Moreover, accumulation of water produced in the reaction onto the biocatalyst causes a decrease in the catalytic activity. As a result, improvements in both reaction rate and final conversion can be achieved by operating in an adsorptive-reactor mode. Control of water in the reactor is achieved with a catalytically inert ion-exchange resin in Na-form. The resin prevents an excessive accumulation of water on the biocatalyst and reduces equilibrium limitations. The thermodynamic activity of water is identified as a key parameter for the design of such reactors. A mathematical model capable of predicting the water activity as a function of the varying concentrations of reactants and products is thus developed and found to successfully predict the experimental behavior observed in laboratory reactors. Substantial improvements in performance predicted by the model are seen experimentally in batch reactions and during the transient operation of continuous-flow fixed-bed reactors combining adsorptive and catalytic functions. 相似文献
4.
Rachida Kirdi Najla Ben Akacha Yosra Messaoudi Mohamed Gargouri 《Biotechnology and Bioprocess Engineering》2017,22(4):413-422
In this work, the transesterification reaction of isoamyl alcohol obtained from fusel oil and leading to the synthesis of isoamyl acetate was conducted simultaneously with in situ ethanol removal, which allows to shift the reaction equilibrium toward ester synthesis. The extracellular Aspergillus oryzae lipase was immobilized into calcium alginate. Effects of immobilization conditions on the loading efficiency and on the specific activity of entrapped lipase were investigated. The kinetic transfer of volatile reactants from the reactor was investigated using an experimentally first order kinetic model, in order to approve the feasibility of the liquid-gas system with continuous ethanol removal in the ester synthesis. The effects of the most influent parameters affecting the reaction have been also investigated using a Doehlert matrix design. The better operating conditions for isoamyl acetate synthesis were: a temperature of 68.5°C and a respective isoamyl alcohol and A. oryzae lipase concentration of 0.72 M and 2.39 g/L. At these conditions, the resulting reaction conversion and ethanol extraction yields were of 89.55 and 69.60%, respectively. The use of the fluidized bed reactor with continuous ethanol removal has allowed to improve the reaction conversion which was two times than the conversion higher obtained in batch reactor. Furthermore, under the optimized conditions in the fluidized bed reactor, the reaction conversion and the ethanol extraction yields were increased by 44.8 and 36.2%, respectively. 相似文献
5.
Claudino MJ Soares D Van Keulen F Marques MP Cabral JM Fernandes P 《Bioresource technology》2008,99(7):2304-2311
Silicone rubbers are hydrophobic, a feature that may prove advantageous if this material is to be used as immobilization matrix in bioconversion systems where hydrophobic species are present, such as sterols and mycobacterial cells. Mycobacterium sp. cells with sitosterol side chain cleavage activity were accordingly effectively adsorbed onto silicone and the potential application of the concept was assessed by matching the behavior of the resulting immobilized biocatalyst with free cells and Celite immobilized cells. Mass transfer, kinetics, thermal and storage stability characterization of a biotransformation system based in the use of the silicone immobilized biocatalyst was performed. The feasibility of biocatalyst reutilization was tentatively explored. 相似文献
6.
7.
The affinity chromatographic reactor (ACR) is a bioreactor which utilizes the dynamic interaction or the dynamic affinity between a free coenzyme and immobilized enzymes for the highly efficient regeneration of dissociable coenzymes. Dynamic affinity between free NAD and immobilized alcohol dehydrogenase (ADH) in ACR was investigated by three different methods. ADH catalyzed both oxidation and reduction of NAD, consuming propionaldehyde and ethanol. The theoretical model under consideration elucidated a criterion for the expression of the dynamic affinity as a relationship among the affinity constants and the concentrations of a coenzyme and immobilized enzyme. This criterion was confirmed experimentally by the measurements of the retention time of NAD and the half-life period of the reactor activity after one-shot pulse injection of NAD to ACR. In the stability measurement of the immobilized enzyme, it became clear that ADH was more stable at the higher concentration in immobilization. Although the present case of coenzyme cycling by a single enzyme is very special, with limited chance for the direct application, the results obtained here provide a theoretical basis for ACR with multienzymes-which is of more general use. 相似文献
8.
In Part I of this series,(1) we derived a model and made simulations for a multistage fluidized bed reactor (MFBR). It was concluded that the MFBR can be an attractive alternative for a fixed bed reactor when operated with a deactivating biocatalyst. In Part II of this series, the design of a laboratory-scale MFBR and its evaluation to investigate the practical feasibility of this reactor type, will be described. Experiments with a duration as long as 10 days were carried out successfully using immobilized glucose isomerase as a model reaction system. The results predicted by the model are in good agreement with the measured glucose concentration and biocatalyst activity gradients, indicating perfect mixing of the particles in the reactor compartments.The diameters of the biocatalyst particles used in the experiments showed a large spread, with the largest being 1.7 times the smallest. Therefore, an additional check was carried out, to make sure that the particles were not segregating according to size. Particles withdrawn from the reactor compartments were investigated using an image analyzer. Histograms of particle size distribution do not indicate segregation and it is concluded that the particles used have been mixed completely within the compartments. As a result, transport of biocatalyst is nearly plug flow. 相似文献
9.
Kuyukina MS Ivshina IB Gavrin AY Podorozhko EA Lozinsky VI Jeffree CE Philp JC 《Journal of microbiological methods》2006,65(3):596-603
A simple biosurfactant-based hydrophobization procedure for poly(vinyl alcohol) (PVA) cryogels was developed allowing effective immobilization of hydrocarbon-oxidizing bacteria. The resulting partially hydrophobized PVA cryogel granules (granule volume 5 microl) contained sufficient number (6.5 x 10(3)) of viable bacterial cells per granule, possessed high mechanical strength and spontaneously located at the interface in water-hydrocarbon system. Such interfacial location of PVA granules allowed high contact of immobilized biocatalyst with hydrophobic substrate and water phase, thus providing bacterial cells with mineral and organic nutrients. As a result, n-hexadecane oxidation efficiency of 51% after 10-day incubation was achieved using immobilized biocatalyst. PVA cryogels with increased hydrophobicity can be used for immobilization of bacterial cultures performing oxidative transformations of water-immiscible organic compounds. Immobilization of in situ biosurfactant producing Rhodococcus bacteria into PVA cryogel is discussed. PVA cryogel granules with entrapped alkanotrophic rhodococcal cells were stable after 10-month storage at room temperature. 相似文献
10.
Direct esterifications using a nylon-immobilized lipase from Candida cylindracea were carried out in batch and continuous-flow reactors. The immobilized enzyme was effective in catalyzing the synthesis of ethylpropionate, isoamylpropionate, and isoamylbutyrate. With ethanol dissolved in hexane as a substrate, the maximum initial esterification rate was 0.02 mole/(h x g of immobilized protein), but the enzyme was stable only when the substrate concentrations were lower than 0.2 M. With isoamyl alcohol in hexane as a substrate, esterification rates as high as 0.085 mole/(h x g of immobilized protein) were observed and the immobilized enzyme was stable over a much broader concentration range. However, in this case, the use of a solvent, such as hexane, was not necessary for esterification, and the enzyme could be employed in equimolar acid/alcohol mixtures. A packed-bed reactor was operated successfully for the continuous synthesis of esters. The reactor was stable for long periods of time, and the steady-state performance could be accurately predicted on the basis of batch reaction experiments. 相似文献
11.
D. T. Mirzarakhmetova M. M. Rakhimov S. Kh. Abdurazakova Z. R. Akhmedova 《Applied Biochemistry and Microbiology》2006,42(2):150-155
The behavior of intact and immobilized invertase in aqueous and water-organic media has been studied. In a water-organic medium, the transferase properties of the enzyme changed: pH optima of intact and immobilized invertase undergo shifts of 0.5 units; the temperature optimum decreases (50°C and 25°C in aqueous and water-organic media, respectively). The extent of conversion of isoamyl alcohol in water-organic medium shows a dependence on several factors (the enzyme and substrate concentrations; amount of the organic phase; and duration of the enzyme incubation with the substrate). Optimum parameters of isoamyl alcohol conversion were used for transforming fusel alcohols into alkyl fructosides. The results of this applied research have important practical implications (conversion of fusel oils of alcoholic beverages). 相似文献
12.
Mirzarakhmetova DT Rakhimov MM Abdurazakova SKh Akhmedova ZR 《Prikladnaia biokhimiia i mikrobiologiia》2006,42(2):169-174
The behavior of intact and immobilized invertase in aqueous and water-organic media has been studied. In a water-organic medium, the transferase properties of the enzyme changed: pH optima of intact and immobilized invertase undergo shifts of 0.5 units: the temperature optimum decreases (50 degrees C and 25 degrees C in aqueous and water-organic media, respectively). The extent of conversion of isoamyl alcohol in water-organic medium shows a dependence on several factors (the enzyme and substrate concentrations; amount of the organic phase; and duration of the enzyme incubation with the substrate). Optimum parameters of isoamyl alcohol conversion were used for transforming fusel alcohols into alkyl fructosides. The results of this applied research have important practical implications (conversion of fusel oils of alcoholic beverages). 相似文献
13.
A lumped model for cell growth and secondary metabolite production in an immobilized live cell bioreactor has been developed. This model is applied here to simulate the performance of an immobilized bioreactor under steady-state conditions and under conditions of periodically varying concentration of a growth-limiting substrate. The results of the simulation study were experimentally verified in the case of the production of the antibiotic candicidin by Streptomyces griseus in an immobilized bioreactor with forced periodic operation. The results of the studies suggest that periodically operated immobilized live cell bioreactors can provide a potent alternative for the production of non-growth-associated biochemicals, as compared to free cell fermentations, pulsed fermentations with process cycle regeneration, and nonregenerated bioreactors. This work has demonstrated that by frequent pulsing of the growth limiting nutrient, stable extended production can be obtained at high specific cellular productivities. 相似文献
14.
Marija??orovi? Mladen?Mihailovi? Katarina?Banjanac Milica?Carevi? Ana?Milivojevi? Nenad?Milosavi? Dejan?Bezbradica
The aim of this study was to develop simple and efficient method for immobilization of Candida antarctica lipase B onto hydrophobic anion exchange resin Purolite® MN102 and to apply immobilized catalyst for the enzymatic synthesis of two valuable esters—isoamyl acetate and l-ascorbyl oleate. At optimized conditions (1 M phosphate buffer pH = 7, 7 h at 25 °C, and 18.75 mg of offered proteins g?1 of support), immobilized lipase with hydrolytic activity of 888.4 p-nitrophenyl butyrate units g?1 was obtained. Afterwards, preparation was applied for the solvent-free synthesis of isoamyl acetate from triacetin and isoamyl alcohol. At 75 °C, 1 M of isoamyl alcohol, and 6 mg ml?1 of enzyme 100 % yield was achieved in 6 h, while at prolonged reaction times, complete conversion was enabled even at lower temperatures, lower lipase loadings, and higher substrate concentrations. After 15 consecutive reuses (60 h), activity of catalyst dropped to 50 % of its initial value and total amount of 1.31 mol (170.55 g) of ester with 1 g of biocatalyst was produced. Even higher operational stability of lipase (25 % loss of activity in 200 h) was observed in the synthesis of l-ascorbyl oleate performed in organic solvent (t-butanol). Multiple use of one batch of immobilized biocatalyst in both cases led to a significant process cost reduction and substantial increment of corresponding productivities. 相似文献
15.
For the application of immobilized enzymes, fixed bed reactors are used almost exclusively. Fixed bed reactors have specific disadvantages, especially for processes with a deactivating catalyst. Therefore, we have studied a novel reactor type with continuous transport of the immobilized biocatalyst. Flow of biocatalyst is countercurrent to the substrate solution. Because of a stagewise reactor design, back-mixing of biocatalyst is very limited and transport is nearly plug flow. The reactor operates at a constant flow rate and conversion, due to constant holdup of catalytic activity. The reactor performance is compared with a configuration of fixed bed reactors. For reactions in the first-order regime, enzyme requirements in this new reactor are slightly less than for fixed bed processes. The multistage fluidized bed appears to be an attractive reactor design to use biocatalyst to a low residual activity. However, nonuniformity of the particles might affect plug flow transport of the biocatalyst. A laboratory scale reactor and experiments are described in Part II(1) of this series. Hydrodynamic design aspects of a multistage fluidized bed are discussed in more detail in Part III.(2). 相似文献
16.
Yoshida H Hara KY Kiriyama K Nakayama H Okazaki F Matsuda F Ogino C Fukuda H Kondo A 《Applied microbiology and biotechnology》2011,91(4):1001-1006
We developed a novel enzymatic glutathione (GSH) production system using Saccharomyces cerevisiae as a whole-cell biocatalyst, and improved its GSH productivity by metabolic engineering. We demonstrated that the metabolic
engineering of GSH pathway and ATP regeneration can significantly improve GSH productivity by up to 1.7-fold higher compared
with the parental strain, respectively. Furthermore, the combination of both improvements in GSH pathway and ATP regeneration
is more effective (2.6-fold) than either improvement individually for GSH enzymatic production using yeast. The improved whole-cell
biocatalyst indicates its great potential for applications to other kinds of ATP-dependent bioproduction. 相似文献
17.
F.M. Plieva K.A. Kochetkov Ishwar Singh V.S. Parmar Yu.N. Belokon' V.I. Lozinsky 《Biotechnology letters》2000,22(7):551-554
Hog pancreas lipase was covalently attached to the beads of poly(vinyl alcohol)-cryogel – a macroporous hydrogel prepared by means of freeze-thaw technique. The immobilized biocatalyst thus obtained was examined in the reaction of enantioselective hydrolysis of the ethyl ester of N-benzylidene derivative of DL-phehylalanine in the medium of acetonitrile (contained 5 vol.% of water without any buffers). Eighty-three %-enantiomeric excess of the l-amino acid was reached after 144 h. Virtually the same result was obtained in the repeated use of the same immobilized biocatalyst after its 6-months-storing in a refrigerator. 相似文献
18.
Ethylglucoside monooleate was synthesized by esterification between ethylglucoside and oleic acid with immobilized lipase from Candida antarctica in a solvent-free system. It was shown that a stirred tank reactor was suitable for the enzymatic reaction process involving substrates with low miscibility, in which the biocatalyst was recycled five times without significant activity loss. Removal of the co-product, water, from the reaction medium by carrying out the reaction under reduced pressure benefited the esterification reaction and increased the monooleate yield up to 97% within 8 hours. 相似文献
19.
A. Krastanov 《Applied microbiology and biotechnology》1997,47(5):476-481
A novel immobilized biocatalyst with invertase activity was prepared by adhesion of yeast cells to wool using glutaraldehyde.
Yeast cells could be immobilized onto wool by treating either the yeast cells or wool or both with glutaraldehyde. Immobilized
cells were not desorbed by washing with 1 M KCl or 0.1 M buffers, pH 3.5–7.5. The biocatalyst shows a maximum enzyme activity
when immobilized at pH 4.2–4.6 and 7.5–8.0. The immobilized biocatalyst was tested in a tubular fixed-bed reactor to investigate
its possible application for continuous full-scale sucrose hydrolysis. The influence of temperature, sugar concentration and
flow rate on the productivity of the reactor and on the specific productivity of the biocatalyst was studied. The system demonstrates
a very good productivity at a temperature of 70 °C and a sugar concentration of 2.0 M. The increase of the volume of the biocatalyst
layer exponentially increases the productivity. The productivity of the immobilized biocatalyst decreases no more than 50%
during 60 days of continuous work at 70 °C and 2.0 M sucrose, but during the first 30 days it remains constant. The cumulative
biocatalyst productivity for 60 days was 4.8 × 103kg inverted sucrose/kg biocatalyst. The biocatalyst was proved to be fully capable of continuous sucrose hydrolysis in fixed-bed
reactors.
Received: 8 November 1996 / Received revision: 31 January 1997 / Accepted: 31 January 1997 相似文献
20.
A kinetic study of isoamyl acetate synthesis by immobilized lipase-catalyzed acetylation in n-hexane 总被引:1,自引:0,他引:1
The objective of this work was to propose a reaction mechanism and to develop a rate equation for the synthesis of isoamyl acetate by acylation of the corresponding alcohol with acetic anhydride using the lipase Novozym 435 in n-hexane. The reaction between isoamyl alcohol and acetic anhydride occurred at high rate in first place. Then, if excess alcohol was used, produced acetic acid further reacted with remaining alcohol, leading to yields higher than 100% (based on initial acetic anhydride content). This reaction was much slower and took place only when acetic anhydride had been totally consumed. Optimal pH for Novozym 435 was 7.7. Acetic acid strongly inactivated the enzyme but it was partially caused by the pH drop in the biocatalyst aqueous microenvironment. Acetic anhydride also showed an important inhibition effect. On the contrary, isoamyl alcohol and isoamyl acetate had no negative effect on the lipase. The analysis of the initial rate data showed that reaction followed a Ping-Pong Bi-Bi mechanism with inhibition by acetic anhydride. The kinetic constants were obtained by multiple regression analysis of experimental findings. Equation predictions and experimental reaction rate values matched very well at conditions where acetic acid concentration in the medium was low. 相似文献