Abstract
A new and cheap support, vermiculite was successfully used to immobilize neutral protease by adsorption and hexamethylene diamine mediated coupling using glutaraldehyde as a bifunctional agent. Neutral protease immobilized on vermiculite by adsorption showed maximum retained activity than HMD mediated coupling. The optimum temperature for both free and immobilized neutral protease was found to be 45°C. However, the pH and thermal stabilities of immobilized neutral protease was observed to be better than that of the free enzyme. The storage stability of the immobilized enzyme was also studied.
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Pelczar, M. J.; Chan, E. C. S.; Krieg, N. R.: Microbiology of soil. In: Microbiology, pp. 543–549. New York: McGraw Hill International Edition 1989
Perez-Mateos, M.; Rad, J. C.: Immobilization of alkaline phosphatase by soil structural units. Biotechnol. Appl. Biochem. 11 (1989) 371–378
Kiss, S.; Dragem-Bularda, M.; Radulescu, D.: Soil polysaccharides: Activity and agricultural importance: In: Burnes, R. G. (Ed): Soil Enzymes, pp. 117–147. London, Academic Press 1978
Sarkar, J. M.; Burns, R. G.: Synthesis and properties of β-D-glucosidase-phenolic copolymers as analogues of soil humic-enzyme complexes. Soil Biol. Biochem. 16 (1984) 619–625
Ladd, J. N.; Paul, E. A.: Changes in enzymic activity and distribution of acid-soluble amino acid nitrogen in soil during nitrogen immobilization and mineralization. Soil Biol. Biochem. 5 (1973) 825–840
Chibata, I.: Preparation of immobilized enzymes and microbial cells: In: Immobilized Enzymes, pp. 11–15. New York: Kodansha Tokyo, J. Wiley & Sons 1978
Kennedy, J. F.: Immobilized enzymes: In: Scouten, W. H. (Ed.): Solid Phase Biochemistry, pp. 265–273. New York: J. Wiley & Sons 1983
Tarafdar, J. C.; Chhonkar, P. K.: Urease clay interactions: I — Adsorption of urease on clays saturated with different cations. J. Indian. Soc. Soil Sci. 30 (1982) 27–32
Sarkar, J. M.; Leonowice, A.; Jean, M.: Immobilization of enzymes on clays and soils. Soil Biol. Biochem. 21 (1989) 223–230
Jackson, M. L.: Phosphorus determinations for soils: In: Soil Chemical analysis, pp. 57–167, New Delhi: Prentice-Hall of India Pvt. Ltd. 1967
Anson, M. L.: The estimation of pepsin, trypsin, papain and cathepsin with haemoglobin. J. Gen. Physiol. 22 (1938) 79–89
Lowry, O. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J.: Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193 (1951) 265–275
Zaborsky, O.: Properties of covalently bonded water-insoluble enzyme-polymer conjugates. In: Immobilized Enzymes, pp. 49–50, Cleaveland, Ohio: C.R.C. Press 1974
Beddows, C. G.; Mirauer, R. A.: Immobilization of β-galactosidase and other enzymes onto p-amino-carbanilated cellulose derivatives. Biotechnol. Bioeng. 22 (1980) 311–321
Axen, R.; Myrin, P. A.: Janson, J. C.: Chemical fixation of chymotrypsin to water insoluble cross linked dextran and solubilization of the enzyme derivative by means of dextranase. Biopolymers 9 (1970) 401–413
Garcia III, A.; Oh, S.; Engler, C. R.: Cellulase immobilization on Fe3O4 and characterization. Biotechnol. Bioeng. 33 (1989) 321–326
Ohmiya, K.; Tanimura, S.; Kobayashi. T.; Shimizu, S.: Preparation and properties of proteases immobilized on anion exchange resin with glutaraldehyde. Biotechnol. Bioeng. 20 (1978) 1–15
Puvanakrishnan, R.; Bose, S. M.: Studies on immobilization of trypsin on sand. Biotechnol. Bioeng. 22 (1980) 919–928
Alvesda Silva, M.; Gill, M. H.: Graft copolymers as supports for the immobilization of biological compounds: In: Guilbault, G. G.; Mascini, M. (Eds): Analytical uses of immobilized biological compounds for detection, medical and industrial uses, pp. 177–185, D. Reidel Publishing Co.
Axen, R.; Ernbach, S.: Chemical fixation of enzymes on cyanogen halide activated polysaccharide carriers. Eur. J. Biochem. 18 (1971) 351–360
Goldstein, L.; Levin, Y.; Katchalski, E.: A water insoluble polyanionic derivative of trypsin. II. Effect of the poly electrolyte carrier on the kinetic behaviour of the bound trypsin. Biochemistry 3 (1964) 1913–1919
Goldstein, L.: A new polyamine carrier for immobilization of proteins water insoluble derivatives of pepsin and trypsin. Biochim. Biophys. Acta. 327 (1973) 132–137
Puvanakrishnan, R.; Bose, S. M.: Immobilization of pepsin on sand: Preparation, characterization, and application. Indian J. Biochem. Biophys. 21 (1984) 323–326
Lozano, P.; Manjon, A.; Romojaro, F.; Iborra, J. L.: Properties of pectolytic enzymes covalently bound to nylon for apricot juice clarification. Process Biochem 23 (1988) 75–78
Krysteva, M. A.; Shopova, B. I.; Yotova, L. Y.; Karasavova, M. I.: Covalent binding of enzymes to synthetic membranes containing acrylamide units, using formaldehyde. Biotechnol. Appl. Biochem. 13 (1991) 106–111
Tarhan, L.; Uslan, A. H.: Characterization and operational stability of immobilized catalase. Process Biochem. 25 (1990) 14–18
Ulbrich, R.; Schellenberger, A.; Damerau, W.: Studies on the thermal inactivation of immobilized enzymes. Biotechnol. Bioeng. 28 (1986) 511–522
Hayashi, T.; Ikada, Y.: Lipoprotein lipase immobilization onto polyacrolein microspheres. Biotechnol. Bioeng. 36 (1990) 593–600
Sreenivasulu, S.; Dhar, S. C.; Puvanakrishnan, R.: Effect of immobilization on the stability and characteristics of alkaline proteinase. Leather Science 32 (1985) 75–80
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Chellapandian, M., Sastry, C.A. Vermiculite as an economic support for immobilization of neutral protease. Bioprocess Eng. 8, 27–31 (1992). https://doi.org/10.1007/BF00369260
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DOI: https://doi.org/10.1007/BF00369260