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1.
Summary Laccase purified from Trametes versicolor oxidizes 2,6-dimethoxyphenol (2,6-DMP) and syringaldazine in hydrophobic solvents presaturated with water, and in hydrophilic organic solvents provided that a sufficient amount of water is added. Ease of performance of the laccase test in organic solvents is improved after immobilization of the enzyme by entrapping in Sepharose CL-6B during enzyme filtration through the gel beads. The gel-enzyme association has been shown to be stable in water-presaturated solvents. Efficiency of the immobilized laccase in organic solvents containing 7% water was 10%–20% of that in potassium-citrate buffer. Immobilized laccase in organic solvents showed good stability and high tolerance to elevated temperatures. 相似文献
2.
Esterification of organic acids and alcohols in aqueous media is very inefficient due to thermodynamic constraints. However, fermentation processes used to produce organic acids and alcohols are often conducted in aqueous media. To produce esters in aqueous media, biphasic alginate beads with immobilized lipase are developed for in situ esterification of butanol and butyric acid. The biphasic beads contain a solid matrix of calcium alginate and hexadecane together with 5 mg/mL of lipase as the biocatalyst. Hexadecane in the biphasic beads serves as an organic phase to facilitate the esterification reaction. Under optimized conditions, the beads are able to catalyze the production of 0.16 mmol of butyl butyrate from 0.5 mmol of butyric acid and 1.5 mmol of butanol. In contrast, when monophasic beads (without hexadecane) are used, only trace amount of butyl butyrate is produced. One main application of biphasic beads is in simultaneous fermentation and esterification (SFE) because the organic phase inside the beads is very stable and does not leach out into the culture medium. SFE is successfully conducted with an esterification yield of 6.32% using biphasic beads containing iso-octane even though the solvent is proven toxic to the butanol-producing Clostridium spp. 相似文献
3.
A hydrophobic silicone polymer could be effectively applied to immobilization of two kinds of biocatalysts operating in organic media. Horse liver alcohol dehydrogenase, which was solubilized in a small amount of water, or deposited on water-filled hydrophilic particles, was immobilized in this material. This configuration of the preparation involving finely dispersed aqueous phase permitted a simple packed-bed operation for the enzymatic oxidation of alcohol and reduction of aldehyde with a coupled-substrate NAD(H) recycling in n-hexane. Another example was the immobilization of Nocardia corallina which catalysed epoxidation of liquid alkenes such as 1-tetradecene, 1-octene, and styrene in the presence of n-hexadecane. In order to adjust the hydrophobicity-hydrophilicity balance of the support, it was effective to immobilize the cells in a mixed matrix composed of silicone polymer and Ca-alginate gel. The optimum composition of the mixed matrix, which yielded the highest productivity of epoxide, was 80-90% silicone + 20-10% alginate for the production of 1,2-epoxytetradecane, 40-50% silicone + 60-50% alginate for 1,2-epoxyoctane, and almost 0% silicone + 100% alginate for styrene oxide. This significant change of the optimum composition was primarily associated with the degree of substrate inhibition. 相似文献
4.
Fifty different hydrolases were screened for retention of high esterification activity in an organic solvent with citronellol as substrate. Although 22 hydrolases were very active as catalysts in the organic solvent, lipase from Candida cylindracea (lipase OF 360) was selected for further examination of the effects of reaction conditions on enzyme activity, with regard to catalyst availability and activity retention after immobilization. When the enzyme was entrapped in hydrophobic polyurethane gels, water-saturated isooctane was found to be the most suitable solvent, whereas polar solvents caused reversible catalyst inactivation. Entrapment significantly enhanced the operational stability of the lipase in the organic solvent. 相似文献
5.
Summary The direct, lipase-catalyzed esterification of hydrophilic diols in organic solvents was achieved by first adsorbing the hydrophilic, solvent immiscible substrate onto a solid support with high internal surface, namely silica gel and reacting the solid mixture with fatty acid vinyl esters in an appropriate organic solvent and in presence of an immobilized lipase fromMucor miehei (Lipozyme). Quantitative conversions of the acyl donors and very high reaction rates were observed in these transformations. Furthermore, mono- or diesters of these diols could be selectively produced by this method. 相似文献
6.
The catalytic feature of serine proteases for synthetic reactions in hydrophilic organic solvents and effects of immobilization by complexation with polysaccharides are described. Free alpha-chymotrypsin and subtilisin Carlsberg catalyze esterification, transesterification, and peptide synthesis in hydro-organic cosolvents with less than 10% water. Subtilisin BPN' is catalytically less active. The medium effects on the reaction kinetics and product yield were investigated in terms of the nature of solvent and water content in the reaction systems. The substrate- and stereo-specificities of the enzymes suggest that the enzymes maintain their native conformations in these low-water organic solvents. The catalytic activities of the proteases markedly increase by immobilization or complexation with polysaccharides, such as chitin or chitosan. The results of the rate measurements suggest that the primary role of the support materials is the activation of the enzymes and the increase in substrate concentration at reaction sites. 相似文献
7.
Li YG Xing JM Xiong XC Li WL Gao HS Liu HZ 《Journal of industrial microbiology & biotechnology》2008,35(3):145-150
The immobilization of Pseudomonas delafieldii R-8 in calcium alginate beads has been studied in order to improve biodesulfurization activity in oil/water (O/W) biphasic
systems. A gas jet extrusion technique was performed to produce immobilized beads. The specific desulfurization rate of 1.5 mm
diameter beads was 1.4-fold higher than that of 4.0 mm. Some nonionic surfactants can significantly increase the activity
of immobilized cells. The desulfurization rate with the addition of 0.5% Span 80 increased 1.8-fold compared with that of
the untreated beads. The rate of biodesulfurization was markedly enhanced by decreasing the size of alginate beads and adding
the surfactant Span 80, most likely resulting from the increasing mass transfer of substrate to gel matrix. 相似文献
8.
《Biocatalysis and Biotransformation》2013,31(1-4):341-351
A novel procedure for attaching lipase to certain kinds of hydrophobic surfaces is described. The procedure involves covalent derivatization of the protein molecule by reaction in solution with a hydrophobic imidoester, aldehyde or activated polyethylene glycol. The resulting protein derivative is then allowed to adsorb onto an insoluble hydrophobic surface. Quantitative adsorption is observed and the enzyme is bound very strongly on the support The number and nature of the hydrophobic substituents introduced in the chemical derivatization step can be easily controlled. The adsorption step occurs spontaneously upon exposure of the modified protein to a variety of hydrophobic materials. The hydrophobic lipase derivative obtained by reaction with PEG activated with p-nirrophenyl chloroformate, for example, adsorbs readily onto polyacrylate and polystyrene beads, with most of its esterification activity in organic solvent intact. Its thermostability is also greatly enhanced. Derivatization of lipase with hydrophobic groups greatly enhances its esterification activity in organic solvent, and its immobilization in this manner enables the preparation of a highly reactive biocatalyst for biotechnological application. 相似文献
9.
alpha-Chymotrypsin (CT), subtilisin BPN' (STB), and subtilisin Carlsberg (STC) were immobilized by adsorption to porous chitosan beads (Chitopearl, CP). The immobilized enzymes showed higher catalytic activities than free enzymes for amino acid esterification in many hydrophilic organic solvents except for methanol and DMF. In ethanol, the initial rate of the esterification increased with water content, whereas in ethyl acetate, the maximum rate was obtained at 2%-3% water. CP-immobilized CT also catalysed transesterification of Ac-Tyr-OMe in ethanol and peptide synthesis in acetonitrile from Ac-Tyr-OH or its ethyl ester and amino acid amides. The immobilized enzymes are highly stable in organic solutions, and can easily be separated from the reaction solutions. Repeated esterifications of Ac-Tyr-OH in acetonitrile by a CP-immobilized CT gave almost constant yields of the ester for more than 3 weeks. 相似文献
10.
《Journal of Fermentation and Bioengineering》1989,67(2):92-96
Different factors which affect the stability of calcium alginate gel beads entrapping viable cells during fermentation were investigated. It was found that among others, the initial population of cells per ml of gel beads, the length of period of incubation in CaCl2 solution, and the concentration of sodium alginate used for the immobilization were the most important factors affecting the stability of the gel beads during fermentation. By using an initial cell population of about 105 cells per ml of 2.0% sodium alginate, and incubating the beads for at least 22 h in a CaCl2 solution after immobilization, the percentage of beads which developed cracks during fermentation was highly reduced. Also, without the addition of CaCl2 into the fermenting broth, the gel beads were stable for nine consecutive batch fermentations. 相似文献
11.
High-molecular weight pectic acid with a STAUDINGER index of 210 ml/g and a degree of esterification of 3%was used as matrix material for the immobilization of Saccharomyces cerevisiae cells. In discontinuous and continuous fermentation tests the gel beads obtained exhibited the same biomass loading capacity (152–155 g dry wt. cells/kg gel) and about the same maximum specific productivity (103.0 g ethanol/kg gel · h) as alginate immobilizates. But there were distinct differences in the swelling behaviour of the two gels. Under the same experimental conditions the increase of bead volume amounted to 27% only for pectate gel in comparison to 129% for alginate gel. In continuous fermentation experiments performed in a horizontal-column packed-bed reactor with liquid recycling a mean steady-state ethanol concetration of 69.1 g/l and a mean productivity of 24.7 g ethanol/lh could be kept constant over a period of more than 10 days. 相似文献
12.
Using free and immobilized whole cells of Pichia pastoris, the biocatalytic oxidation of benzyl alcohol was investigated in different two-phase systems. This reaction was strongly influenced by both the substrate and product inhibitions, and the production rate of benzaldehyde in the aqueous system became maximum at the initial substrate concentration of ca. 29 g/L with the aldehyde formation less than 4 to 5 g/L even after a longer reaction period. The reaction rates in the two-liquid phase systems were predominantly determined by the partitioning behaviors of the substrate and product between the two phases rather than by enzyme deactivation by the organic solvents. In the two-liquid phase systems, consequently, the organic solvent acted as a reservior to reduce these inhibitory effects, and it was essential to select the organic solvent providing the optimal partitioning of the substrate into the aqueous phase as well as the preferential extraction of the product into the organic phase. The whole cells immobilized in a mixed matrix composed of silicone polymer [>50% (v/v)] and Ca alginate gel (<50%) worked well in the xylene and decane media, providing comparable activities with the free cells. The production rate of aldehyde was also influenced by the solute partitioning into the hydrophilic alginate phase where the cells existed. (c) 1994 John Wiley & Sons, Inc. 相似文献
13.
Kyung Hwa Lee Chang-Ho Park Eun Yeol Lee 《Bioprocess and biosystems engineering》2010,33(9):1059-1065
Glycerol carbonate was synthesized from renewable glycerol and dimethyl carbonate using lipase in solvent-free reaction system
in which excess dimethyl carbonate played as the reaction medium. A variety of lipases have been tested for their abilities
to catalyze transesterification reaction, and Candida antartica lipase B and Novozyme 435 exhibited higher catalytic activities. The silica-coated glycerol with a 1:1 ratio was supplied
to prevent two-phase formation between hydrophobic dimethyl carbonate and hydrophilic glycerol. Glycerol carbonate was successfully
synthesized with more than 90% conversion from dimethyl carbonate and glycerol with a molar ratio of 10 using Novozyme 435-catalyzed
transesterification at 70 °C. The Novozyme 435 [5% (w/w) and 20% (w/w)] and silica gel were more than four times recycled
with good stability in a repeated batch operation for the solvent-free synthesis of glycerol carbonate. 相似文献
14.
Christian Tamponnet François. Coslantino Jean-Noel Barbotin Regis Calvayrac 《Physiologia plantarum》1985,63(3):277-283
The immobilisation of Euglena gracilis Z cells in a calcium alginate matrix maintained respiratory and photosynthetic activities and ultrastructural integrity. Moreover, immobilization did not prevent Euglena cells from greening inside the gel beads. Electron microscopy demonstrated that the immobilized cells were fixed in the same cellular state as they were when the immobilization occurred. This can he explained by simultaneous reaction of both Ca2+ and the alginate with the cells. Some hypotheses about the role of C2+ are discussed. In addition, long term storage (2 years) in calcium alginate has been performed permitting applications in algal storage. 相似文献
15.
The reaction rate of two lipase-catalysed reactions, esterification and transesterification, were studied in a liquid/solid two-phase system in order to investigate the effect of water partition between the enzyme preparation and the liquid phase composed of only the reactants, i.e. without the conventional solvents. Lipase from Candida cylindracea was used for these studies. The enzyme was inactive in dehydrated systems. In the case of monoester synthesis, the reaction rate increased with increasing water activity. The reaction rates of the non-specific C. cylindracea lipase-catalysed reactions were very sensitive to the nature of the substrates in this unusual system. For instance, the transesterification reaction rate of ethyl propionate was 48 times higher with nonanol than heptanol in the case of dehydrated substrates, but only 2.2 times higher in the case of water-saturated substrates. The results presented here demonstrate the absolute necessity to consider the polarity of every substrate, because of its ability to modify the water partition between the solid phase (enzyme preparation) and the liquid phase (substrate and product), which results in drastic changes in enzyme activity. Contrary to esterification, which is known to be activated by the water produced, the rate of transesterification remained constant at the beginning of the reaction. However, when transesterification and esterification were carried out in the same liquid phase, the transesterification reaction rate was controlled by the water produced by the concomitant esterification. Activation effects of the water molecules produced during the enzymatic reaction were of exactly the same order of magnitude for both reactions. 相似文献
16.
Sang Hyun Lee Thanh Thi Ngoc Doan Sung Ho Ha Woo-Jin Chang Yoon-Mo Koo 《Journal of Molecular Catalysis .B, Enzymatic》2007,47(3-4):129-134
The immobilization of lipase from Candida rugosa, using ionic liquids as additives to protect the inactivation of lipase by released alcohol and shrinking of gel during sol–gel process, was investigated. The influence of various factors, such as structure of ionic liquids, content of ionic liquids and types of precursor in the sol–gel process on the activity and stability of immobilized lipase was also studied. The highest hydrolytic activity of immobilized lipase was obtained when the hydrophilic ionic liquid, [C2mim][BF4], was used as an additive, while the highest stability of immobilized lipase was obtained by using hydrophobic ionic liquid, [C16mim][Tf2N]. Therefore, the binary mixtures of these ionic liquids as additives were used to obtain the optimal immobilized lipase, which shows both high activity and stability. The hydrolysis and esterification activities of lipase co-immobilized with the mixture of 1:1 at molar ratio of [C2mim][BF4] and [C16mim][Tf2N] were 10-fold and 14-fold greater than in silica gel without ionic liquids (ILs), respectively. After 5 days incubation of this immobilized lipase in n-hexane at 50 °C, 84% of initial activity was remained, while the residual activity of the lipase immobilized without ILs was 28%. 相似文献
17.
A newly isolated strain of the yeast Saccharomyces cerevisiae is investigated for the biocatalytic reduction of ketones and the oxidation of alcohols in organic solvents. The yeast cells are immobilized by entrapment within calcium alginate beads and are found to possess the ability to stereoselectively reduce prochiral ketones and oxidize chiral alcohols to equilibrium conversions. The effect of reactant partitioning on the initial rate of the reactions is also investigated. The observed initial rates are found to vary inversely with reactant partitioning between the organic solvent and the biocatalyst beads. A kinetic model is developed to describe the initial reaction rate of hexanone reduction as a function of substrate concentration within the alginate beads. 相似文献
18.
Kanwar SS Kaushal RK Sultana H Chimni SS 《Acta microbiologica et immunologica Hungarica》2006,53(1):77-87
An alkaline thermotolerant lipase of Bacillus coagulans BTS1 was successively purified by ammonium sulfate precipitation and DEAE anion exchange chromatography. The purified lipase immobilized in alginate beads showed an optimal activity at pH 7.5 and 55 degrees C. A pH of 5.0 or 10.0 completely quenched the activity of immobilized lipase. The alginate-bound lipase retained its activity following exposure to most of the organic solvents including amines, alkanes and alcohols. Chloride salt of Al3+, Co2+, Mg2+ and NH4+ modulated the lipase activity of alginate-immobilized enzyme. The alginate entrapped lipase showed a preferentially high activity towards p-nitrophenyl palmitate (C: 16) and activity of matrix increased following exposure to SDS. Moreover, the immobilized lipase retained more than 50% of its activity after 3rd cycle of reuse. 相似文献
19.
The use of organic solvents as reaction media for enzymatic reactions has many advantages. Several organic solvents have been proposed as reaction media, especially for transesterifications using Candida antarctica lipase B (CalB). Among organic solvents, tert-butanol is associated with an enhanced conversion rate in bio-diesel production. Thus, it is necessary to understand the effect of tert-butanol on CalB to explain the high-catalytic efficiency compared with the reaction in other hydrophilic organic solvents. In this study, the effects of tert-butanol on the structure of CalB were investigated by MD simulations. The overall flexibility was increased in the presence of tert-butanol. The substrate entrance and the binding pocket size of CalB in tert-butanol were maintained as in TIP3P water. The distance between the catalytic residues of CalB in tert-butanol indicated a higher likelihood of forming hydrogen bonds. These structural analyses could be useful for understanding the effect of tert-butanol on lipase transesterification. 相似文献
20.
Jin Chuan Wu Yew Kong Wong Kiat Wei Chang Chor Yong Tay Yvonne Chow Mmr Talukder Won Jae Choi 《Biocatalysis and Biotransformation》2007,25(6):459-463
Mucor javanicus lipase was entrapped in alginate-silica hybrid gel beads with or without simultaneous cross-linking with glutaraldehyde. The activity and recovery of activity on immobilization of the enzyme entrapped in the hybrid beads were 1.4 and 1.7 times higher than those of the enzyme entrapped in the simple alginate beads. Entrapment with simultaneous cross-linking in the hybrid beads further improved the enzyme activity (1.6 times) and activity recovery (1.7 times) compared to those of the enzyme entrapped in the hybrid beads without simultaneous cross-linking. The leakage of the enzyme entrapped in the hybrid beads with simultaneous cross-linking was only 50% that of the enzyme entrapped in the simple alginate beads. 相似文献