Lipase B from Candida antarctica immobilized on octadecyl Sepabeads: A very stable biocatalyst in the presence of hydrogen peroxide (original) (raw)
Related papers
Covalent immobilization of lipase from Candida rugosa on Eupergit®
Acta periodica technologica, 2005
An approach is presented for the stable covalent immobilization of lipase from Candida rugosa on Eupergit ® with a high retention of hydrolytic activity. It comprises covalent bonding via lipase carbohydrate moiety previously modified by periodate oxidation, allowing a reduction in the involvement of the enzyme functional groups that are probably important in the catalytic mechanism. The hydrolytic activities of the lipase immobilized on Eupergit ® by two conventional methods (via oxirane group and via glutaraldehyde) and with periodate method were compared. Results of lipase assays suggest that periodate method is superior for lipase immobilization on Eupergit ® among methods applied in this study with respect to both, yield of immobilization and hydrolytic activity of the immobilized enzyme.
Streamlined Preparation of Immobilized Candida antarctica Lipase B
ACS Omega
Candida antarctica lipase B (CalB) was efficiently expressed (6.2 g L −1) in Escherichia coli by utilizing an N-terminal tag cassette and the XylS/Pm expression system in a fed-batch bioreactor; subsequent direct binding to EziG from crude extracts resulted in an immobilized catalyst with superior activity to Novozym 435.
Kinetic studies of lipase from Candida rugosa
Applied Biochemistry and Biotechnology, 2001
The search for an in expensive support has motivated our group to undertake this work dealing with the use of chitosan as matrix for immobilizing lipase. In addition to its low cost, chitosan has several advantages for use as a support, including its lack of toxicity and chemical reactivity, allowing easy fixation of enzymes. In this article, we describe the immobilization of Canada rugosa lipase onto porous chitosan beads for the enzymatic hydrolysis of oliveoil. The binding of the lipase onto the support was performed by physicalad sorption using hexane as the dispersion medium. A comparativestudy between free and immobilized lipase was conducted in terms of pH, temperature, and thermal stability. A slightly lower value for optimum pH (6.0) was found for the immobilized form in comparison with that attained for the soluble lipase (7.0). The optimum reaction temperature shifted from 37°C for the free lipase to 50°C for the chitosan lipase. The patterns of heat stability indicated that the immobilization process tends to stabilize the enzyme. The half-life of the soluble free lipase at 55°C was equal to 0.71 h (K d=0.98 h−1), whereas for the immobilized lipase it was 1.10 h (K d=0.63 h−1). Kinetics was tested at 37°C following the hydrolysis of olive oil and obeys the Michaelis-Menten type of rate equation. The K m was 0.15 mM and the V max was 51 μmol/(min·mg), which were lower than for free lipase, suggesting that the apparent affinity toward the substrate changes and that the activity of the immobilized lipase decreases during the course of immobilization.
2011
In spite of their excellent catalytic properties, enzymes should be improved before their implementation both in industrial and laboratorium scales. Immobilization of enzyme is one of the ways to improve their properties. Candida antarctica lipase B (Cal-B) has been reported in numerous publications to be a particularly useful enzyme catalizing in many type of reaction including regioand enantiosynthesis. For this case, cross-linking of immobilized Cal-B with 1,2,7,8 diepoxy octane is one of methods that proved significantly more stable from denaturation by heat, organic solvents, and proteolysis than lyophilized powder or soluble enzymes. More over, the aim of this procedure is to improve the activity and reusability of lipase. Enzyme kinetics test was carried out by transesterification reaction between 4-nitrophenyl acetate (pNPA) and methanol by varying substrate concentrations, and the result is immobilized enzymes follows the Michaelis-Menten models and their activity is match ...
Candida rugosa Lipase: A Traditional and Complex Biocatalyst
Current Organic Chemistry, 2006
Different commercial preparations from Candida rugosa lipase lead often to an irreproducible behaviour when employed in slightly hydrated media, even when different samples from the same supplier are used. This conduct is triggered by several causes, such as the different concentration of the "real" catalyst in the different crude samples (quantity), or the inherent problems related to heterogeneous biocatalysts (i. e., different amount of water, diffusional problems, etc); furthermore, for C. rugosa lipase, the diverse percentage of different isoenzymes (quality) is another irreproducibility-inducing factor. In this sense, for a rational understanding of all the experimental data described for this complex biocatalyst, topics like the description of the role of the inducer on the fermentation, the biochemical characterisation of the crude biocatalysts and the establishment of synthetical strategies to overcome the irreproducibility problems must be covered. Once these tasks are fulfilled, we will be able to understand the relative catalytic activity of different samples from different origins, showing that enzymes should not merely be considered as a "white magic powder" with synthetical utility. Different techniques to evaluate the importance of the amount of water are discussed: 1 H-NMR for comparing the hydrolytic activity of isoenzymes, and sorption isotherms and Thermogravimetric and Differential Thermal Analysis (TGA/DTA) for understanding the esterification in organic media. Furthermore, different characterisation reactions useful for quantifying the lipase loading (heptyl oleate synthesis and transesterification of vinyl acetate with 1-heptanol) are proposed. Finally, a synthetical strategy based on the acylation, via vinyl acetate, of 1heptanol, geraniol, nerol and cyclohexanol, for comparing the catalytical results and the isoenzymatic profile is discussed.
Covalent immobilization of pure isoenzymes from lipase of Candida rugosa
Enzyme and Microbial Technology, 1997
Covalent irnmobilizution of pure lipases A and B from Candida rugosa on agarose and silica is described. The immobilization increases the half-life of the biocatalysts (tx. = 5 h) with respect to the native pure lipases (tl,? = 0.28 h). The percentage immobilization of lipases A and B is similar in both supports (33-40%). The remaining activity of the biocatalysts immobilized on agarose (70-75%) is greater than that of the enzymatic derivatives immobilized on SiO, (40-50%). The surjixce area and the hydrophobic/hydrophilic properties of the support control the lipase activiw of these derivatives. The thermal stability of the immobilized lipase A derivatives is greater than that of lipase B derivatives. The nature of the support influences the thermal deactivation profile of the immobilized derivatives. The immobilization in agarose (hydrophilic support) gives biocatalysts that show a greater initial specific reaction rate than the biocatalysts immobilized in Si02 (hydrophobic support) using the hydrolysis of the esters of(R) or(S) 2-chloropropanoic and of (R,S) 2-phenylpropanoic acids as the reaction test. The enzymatic derivatives are active for at least 196 h under hydrolysis conditions. The stereospecifi+v of the native and the immobilized enzymes is the same.
Enzyme and Microbial Technology, 1999
Covalent immobilization of C. antarctica lipase B (CALB) on sepharose, alumina, and silica was undertaken. The thermal stability of these covalently immobilized catalysts were studied and compared to adsorbed derivatives from Novo Nordisk at 50°C under wet conditions. Native enzyme and Novozym 435 follow a deactivation model E 3 E 1 whereas covalently immobilized derivatives and SP435A follow the model E 3 E 1 3 E 2. This different behavior is related to the nature of the support and the immobilization methodology. Water absorption isotherms of dry solid biocatalysts in air or isooctane were used to predict the optimum preequilibrium a w value to obtain the highest rate in the esterification of (r,s)-ibuprofen.
Medical Research Journal
Lipases are commonly applied in the pharmaceutical and chemical industry, especially in immobilized form. The use of immobilized lipases facilitates the design of reactors and control of reactions, for example, fast stopping the reaction. The immobilization procedure should increase the stability of the lipase and its activity, as well as be simple and efficient. Lipase B from Candida antarctica (CAL-B) is an enzyme from the lipase group, isolated from the Candida antarctica species. CAL-B has the highest activity in non-polar organic solvents, such as hexane and toluene, and the lowest in polar solvents, e.g. acetonitrile. Due to its hydrolytic properties, this enzyme degrades triglycerides of fatty acids to free fatty acids (FFA) and glycerol. Described lipase is often immobilized, in the aim to increase enantioselective and lipolytic activity. The kinetic and dynamic resolution with the application of lipase is one of the ways in obtaining an enantiopure form of the drugs, which usually are more effective and safer for the patient. The CAL-B could be also applied in the kinetic resolution of compounds being building blocks, derivates of drugs or conjugated forms. Furthermore, the CAL-B is used in the reactions in receiving of organic compounds, which are the natural origin, especially vegetable. Based on the presented data, it can be concluded, that CAL-B is an enzyme with a wide application in the biosynthesis of compounds with therapeutic activity.