Immobilization of Papain on the Mesoporous Molecular Sieve MCM‐48
ABSTRACT The immobilization of papain on the mesoporous molecular sieve MCM-48 (with a pore size of 6.2 nm in diameter) with the aid of glutaraldehyde, and the characteristics of this immobilized papain are described. The optimum conditions for immobilization were as follows: 20 mg native free enzyme/g of the MCM-48 and 0.75 % glutaraldehyde, 2 h at 10–20 °C and pH 7.0. Under these optimum conditions for immobilization, the activity yield [%] of the immobilized enzyme was around 70 %. The influence of the pH on the activity of the immobilized enzyme was much lower compared to the free enzyme. The thermostability of the immobilized enzyme, whose half-life was more than 2500 min, was greatly improved and was found to be significantly higher than that of the free enzyme (about 80 min). The immobilized enzyme also showed good operational stability, and the activity of the immobilized enzyme continued to maintain 76.5 % of the initial activity even after a 12-day continuous operation. Moreover, the immobilized enzyme still exhibited good storage stability. From these results, papain immobilized on the MCM-48 with the aid of glutaraldehyde, can be used as a high-performance biocatalyst in biotechnological processing, in particular in industrial and medical applications.
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ABSTRACT: The conventional protocols for in solution or in gel protein digestion require many steps and long reaction times. The use of trypsin immobilized onto solid supports has recently captured the attention of many research groups, because these systems can speed-up protein digestion significantly. The utilization of new materials such as mesoporous silica as supports, in which enzyme and substrate are dramatically concentrated and confined in the nanospace, offers new opportunities to reduce the complexity of proteomics workflows. An overview of the procedures for in situ proteolysis of single proteins or complex protein mixtures is reported, with a special focus on porous materials used as catalysts. The challenging efforts for designing such systems aimed at mimicking the biochemistry of living cells are reviewed. Potentials, limitations and challenges of this branch of enzyme catalysis, which we indicate as in mesopore digestion, are discussed, in relation to its suitability for high-speed and high-throughput proteomics.Molecules 01/2011; 16(7):5938-62. · 2.43 Impact Factor
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ABSTRACT: Extracellular lipase from Bacillus coagulans BTS-3 was immobilized on (3 A x 1.5 mm) molecular sieve. The molecular sieve showed approximately 68.48% binding efficiency for lipase (specific activity 55 IU mg(-1)). The immobilized enzyme achieved approx 90% conversion of acetic acid and 4-nitrophenol (100 mM each) into 4-nitrophenyl acetate in n-heptane at 65 degrees C in 3 h. When alkane of C-chain length other than n-heptane was used as the organic solvent, the conversion of 4-nitrophenol and acetic acid was found to decrease. About 88.6% conversion of the reactants into ester was achieved when reactants were used at molar ratio of 1:1. The immobilized lipase brought about conversion of approximately 58% for esterification of 4-nitrophenol and acetic acid into 4-nitrophenyl acetate at a temperature of 65 degrees C after reuse for 5 cycles.Journal of Industrial Microbiology 01/2009; 36(3):401-7. · 1.80 Impact Factor
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ABSTRACT: The functionalized mesoporous activated carbon (FMAC) was used as support material for immobilization of acid protease (AP). Immobilization of acid protease on functionalized mesoporous activated carbon (AP–FMAC) performs as a suitable enzyme carrier. Under optimized condition pH (6.0) acid protease 150 mg g−1 FMAC has been adsorbed. The optimum temperature for both free and immobilized AP activities was 50 °C. After incubation at 50 °C, the immobilized AP maintained 50% of its initial activity, while the free enzyme was completely inactivated. A significant catalytic efficiency was maintained along for more than five consecutive reaction cycles in AP–FMAC combination immobilized system. The functional groups of the AP, FMAC and AP–FMAC were observed by Fourier transformer infrared spectroscopy (FT-IR). The scanning electron microscopy (SEM) allowed us to observe the morphology of the surface of FMAC and the AP–FMAC.Biochemical Engineering Journal. 01/2009;