Immobilization of Lipase by Entrapment in Ca-alginate Beads (original) (raw)
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Optimization of lipase entrapment in Ca-alginate gel beads
2005
Lipase from Candida rugosa was entrapped by drop-wise addition of an aqueous mixture of sodium alginate and the biocatalyst to a hardening solution of a Ca 2+ salt. Effects of immobilization conditions such as alginate concentration, CaCl 2 concentration, ratio by weight of ...
Immobilization of Lipase from Rhizopus arrhizus by Entrapment in Calcium Alginate Beads
An extracellular lipase producing mutant stain of Rhizopus arrhizus isolated and purified was immobilised in Ca-alginate beads by Gel entrapment process. The alginate beads were prepared as sodium alginate in crude enzyme extract and CaCl2 to increase its reusability and stability. In the presence of Ca +2 cations, the enzyme-alginate suspension was crosslinked to form beads at room temperature. The effects of reaction parameters such as Na-alginate and CaCl2 concentrations were optimized and the corresponding bead sizes were measured for the alginate maximum production of lipase by immobilized enzyme. The optimum concentration of Na-alginate and CaCl2 were found to be 6% (w/v) and 0.4M respectively. The maximum production of immobilized lipase was observed with bead size 1.5mm. After immobilization, 60-% lipase activity was retained. The entrapped lipase was stable over a wide range of temperature (30 0 c – 60 0 c) and pH (5-9.5). The Ca-alginate entrapped fungal lipase can be used in the industry.
2015
Extracellular lipase producing psychrotrophic Pseudomonas ADT3 (NCBI GenBank Acc.no.JX914667) isolated from soil sample of NyAlesund, Svalbard, Arctic region produced maximal lipase activity of 527.8U/mg after 48 hours at pH 8.5 and temperature 22°C in presence of 1.2mM lead as cofactor. It was partially purified 2.9 folds by ammonium sulphate precipitation (80%). Enzymatic performance was improved by immobilization of enzyme on various carriers viz. Alginate and polyacrylamide gel. The immobilization yield of enzyme immobilized in polyacrylamide gel was low (40.0%) in comparison to that immobilized with alginate (70.0%). Different concentrations of alginate and calcium chloride were studied to acquire stable beads. Optimum concentration of alginate and calcium chloride was 2% and 0.12M respectively. The immobilized enzyme was found to be stable in alkaline pH. The maximal activity for immobilized enzyme was found at pH 8.5. Broader pH tolerance could be achieved by immobilization. ...
Immobilization of a Plant Lipase from Pachira aquatica in Alginate and Alginate/PVA Beads
Enzyme Research, 2014
This study reports the immobilization of a new lipase isolated from oleaginous seeds ofPachira aquatica, using beads of calcium alginate (Alg) and poly(vinyl alcohol) (PVA). We evaluated the morphology, number of cycles of reuse, optimum temperature, and temperature stability of both immobilization methods compared to the free enzyme. The immobilized enzymes were more stable than the free enzyme, keeping 60% of the original activity after 4 h at 50°C. The immobilized lipase was reused several times, with activity decreasing to approximately 50% after 5 cycles. Both the free and immobilized enzymes were found to be optimally active between 30 and 40°C.
Gelatin blends with alginate: gels for lipase immobilization and purification
Biotechnology …, 2003
Blends of natural polysaccharide sodium alginate (5%) with gelatin (3%) cross-linked with glutaraldehyde provide beads with excellent compressive strength (8 × 10 4 Pa) and regular structure on treatment with calcium chloride. Lipases from porcine pancreas, Pseudomonas cepacia, and Candida rugosa were immobilized in such a blend with excellent efficiency. The immobilized enzymes were stable and were reused several times without significant loss of enzyme activity both in aqueous and reverse micellar media. The beads were functionalized with succinic anhydride to obtain beads with extra carboxylic acid groups. These functionalized beads were then successfully used for 7.4-fold purification of crude porcine pancreatic lipase in a simple operation of protein binding at pH 5 and release at pH 8.5.
Immobilization of Cellulase Enzyme in Calcium Alginate Gel and Its Immobilized Stability
2013
In this study, cellulase enzyme is immobilized in calcium alginate bead by entrapment method. Sodium alginate concentration used for alginate bead forming is 2%. Immobilized enzyme activity is evaluated on carboxymethyl cellulose (CMC). The maximum efficiency of enzyme immobilization is 83.645% with immobilized time 30 minute and bead diameter 3mm. The optimum pH value of immobilized enzyme and free enzyme is 4.5. The optimum temperature value of immobilized enzyme is higher than free enzyme, 60 o C and 55 o C respectively. Immobilized enzyme is more stable versus the change of pH and temperature of environment than free enzyme. Immobilized enzyme can stand in higher acidity and temperature than free enzyme. Immobilized cellulase could be reused many times. Immobilized enzyme activity remains 69.2% after 5 recycles and still 20.3% after 8 recycles.
Arthrobacter sp. lipase immobilization for improvement in stability and enantioselectivity
Applied microbiology …, 2006
Arthrobacter sp. lipase (ABL, MTCC no. 5125) is being recognized as an efficient enzyme for the resolution of drugs and their intermediates. The immobilization of ABL on various matrices for its enantioselectivity, stability, and reusability has been studied. Immobilization by covalent bonding on sepharose and silica afforded a maximum of 380 and 40 IU/g activity, respectively, whereas sol-gel entrapment provided a maximum of 150 IU/g activity in dry powder. The immobilized enzyme displayed excellent stability in the pH range of 4-10 and even at higher temperature, i.e., 50-60°C, compared to free enzyme, which is unstable under extreme conditions. The resolution of racemic auxiliaries like 1-phenyl ethanol and an intermediate of antidepressant drug fluoxetine, i.e., ethyl 3-hydroxy-3-phenylpropanoate alkyl acylates, provided exclusively R-(+) products (∼99% ee, E=646 and 473), compared to cell free extract/whole cells which gave a product with ∼96% ee (E=106 and 150). The repeated use (ten times) of covalently immobilized and entrapped ABL resulted in no loss in activity, thus demonstrating its prospects for commercial applications.
Journal of Molecular Catalysis B: Enzymatic, 2009
Lipase from Pseudomonas sp. was immobilized in alginate gel beads. The optimum condition for lipase entrapment was a 2% (w/v) alginate concentration, 100 mM CaCl 2 concentration, 30 U/mL enzyme concentration and a 2.03 mm mean bead size. Under these conditions, 8.11 U/mL of immobilized lipase was obtained with 22.2% of retained activity. Palm oil was used as the starting material to produce monoacylglycerol (MAG) in the glycerolysis reaction. The addition of glycerol in the immobilization step improved the yield of MAG. In order to prevent enzyme from leaking out of the gel beads, beads were coated with silicate. The silicate coated beads showed a higher reusability in the glycerolysis reaction compared to non-coated beads. The production of MAG by coated alginate gel beads was optimized. A 10:1 molar ratio of glycerol to palm oil without water addition, 27 U of immobilized enzyme and 50% (w/v) of palm oil in 2-methyl-2-butanol gave the highest MAG production rate of 0.34 M/h while 10% (w/v) of palm oil in 2-methyl-2-butanol gave the highest overall conversion of triacylglycerol 100% to 54% monoacylglycerol, 9% diacylglycerol and 37% free fatty acid after 4 h.
Immobolization of Halkolphylic lipase from bacelius cerus MS6 Bacreria..................
Lipases belong to the class of hydrolases which are able to catalyze the hydrolysis of long chain triglycerides and many other reactions including ester synthesis and transesterification in organic media containing minute concentration of water. The immobilization of enzyme is an important strategy to improve desirable features of enzymes besides enhancing the stability of the enzyme and to obtain additional beneficial properties via immobilization process. In addition, the immobilized enzyme could be used repeatedly or continuously. In our study, the isolated lipase produced by Bacillus cereus MS6 was immobilized on 3% sodiumalginate beads by the entrapment methods. The alginate bead was prepared as an aqueous mixture of sodium alginate, the enzyme and CaCl 2 to increase its reusability, and overall enzyme stability. Various parameters such as alginate and CaCl 2 concentration, lipase concentration loading and bead size were evaluated for the optimum immobilization yield. It was observed that with an increase in alginate concentration, the yield of immobilized enzyme was also increased up to a certain limit. A similar case was observed with CaCl 2 addition, the optimum concentrations of alginate and CaCl 2 were observed to be 3% and 2% (w/v), respectively. The maximum production and activity of immobilized lipase were 180 mg/ml and 550 U/ml, respectively with an observed yield of 88.70 %, while the maximum activity of free enzyme 10 Mohammed Mohammed Abdu Al-ZaZaee et al was 450 U/ml with an activity yield of 40%. An increased beads size resulted in the decrease of the yield of immobilization. These lipase immobilized beads could be reused for many cycles for the hydrolysis of triglycerides without any loss in the activity. The entrapped lipase was more stable over a wide range of temperatures, pH, and storage time as compared to free enzyme. The catalytic properties of the immobilized lipase were also compared with that of the free enzyme. The optimum pH of the free enzyme was found to be 9.0 at a temperature maximum of 40 0 C, while that the immobilized of lipase was pH10.0 at a temperature maximum of 50 0 C. The optimum substrate concentration for free and immobilized enzyme was found to be 2% and 3%, respectively. The overall characterization data reveals an increase in the stability of the immobilized lipase as compared to free lipase.
International Journal of Biological Macromolecules, 2019
Extracellular lipase from Yarrowia lipolytica was immobilized by ionotropic gelation with alginate and chitosan as encapsulating agents. Photomicrographs revealed a collapsed and heterogeneous surface of these microcapsules due to freeze-drying process. The optimum reaction temperature for the microencapsulated lipase (40°C) was higher than for free lipase (35°C) as well as the optimum pH (8.0 and 7.5, respectively). The study of the reaction kinetics showed that a higher maximum reaction rate (V max) (221.1 U/mg) for the free lipase in comparison to the immobilized form (175.3 U/mg). A protective effect of the microcapsule was detected in the storage of the enzyme at room temperature, as after 75 days 35% of activity was maintained for the microcapsules, while no activity remained after 15 days with the free enzyme. Lower values for inactivation constant (k d) and increase in half-life for immobilized lipase showed that lipase microencapsulation favored the thermostability of this enzyme.