Quantitative Analysis of the Degree of Silanization by the Ninhydrin Method and its Application to the Immobilization of GL-7-ACA Acylase and Cellulolytic Enzyme

  • Published : 2001.04.01

Abstract

A simple quantitative method to measure the degree of silanization was developed, based on the reaction of ninhydrin with the silanization reagent (3-aminopropyltriethoxysilane, 3-APTES). At low concentrations (0.001-0.005%, v/v) of 3-APTES, a good linearity was obtained when 3-APTES reacted with undiluted ninhydrin for 30 min. On the other hand, at high levels of 3-APTES, a linearity was obtained when 3-APTES reacted with 3-fold diluted ninhydrin for 20 min. The reliability of regression curves mentioned above was expressed as a regression coefficient ($R^2$) of more than 0.99. Immobilization of different enzymes was introduced via silanization by using the 3-APTES in order to confirm the validity of the ninhydrin method. When yield for each step in the immobilizatio process were compared, yields of both glutaraldehyde and protein were founc to have the same tendency to silanization. These results shw that the ninhydrin method was suitable for quatitative analysis of silanization and that yields of immobilization could be pre-estimated by measuring silanization levels using the ninhydrin method.

Keywords

References

  1. Appl. Biochem. Biotechnol. v.50 Characterization of immobilized catalaxes and their application in pasteurization of milk with H₂O₂ Akertek, E.;L. Tarhan
  2. Anal. Biochem. v.211 Quantitative determination of chitosan and the percentage of free amino groups Curotto, E.;F. Aros
  3. Appl. Biochem. Biotechnol. v.50 Immobilization of thiabendazole-specific monoclonal antibodies to silicon substrates via aqueous silanization Flounders, A. W.;D. L. Brandon;A. H. Bates
  4. J. Microbiol. Biotechnol. v.7 Hydrolysis of olive oil by lipase, immobilized on hydrophobic support Jung, J. Y.;H. S. Yun;E. K. Kim
  5. J. Microbiol. Biotechnol. v.10 Beef tallow hydrolysis by immobilized lipase Kim, D.-J.;D.-H. Shin;B.-K. Hur;E.-K. Kim
  6. Biotechnol. Bioeng. v.26 Immobilization of a soluble chemically thermostabilized enzyme Lenders, J. P.;P. Germain;R. R. Chichton
  7. J. Biol. Chem. v.193 Protein measurement with folin phenol reagent Lowry, O. H.;N. L. Rosebrough;A. L. Farr;R. J. Randall
  8. J. Biol. Chem. v.176 Photometric ninhydrin method for use in the chromatography of amino acid Moore, S.;W. H. Stein
  9. J. Biomed. Mater. Res. v.40 Chemical modification of titanium surfaces for covalent attachment of biological molecules Nanci, A.;J. D. Wuest;L. Peru;P. Brunet;V. Sharma;S. Zalzal;M. D. Mckee
  10. Biotechnol. Bioeng. v.60 Detection of functional groups and antibodies on microfabricated surfaces by confocal microscopy Nashat, A. H.;M. Mornne;M. Ferrari
  11. J. Microbiol. Biotechnol. v.8 Effect of cross-linking agents on L-sorbose production by immobilized Gluconobacter suboxydans cells Park, Y. M.;S. K. Rhee;E. S. Choi;I. S. Chung
  12. Chem. Mikrob. Tech. Lebensm. v.9 Spectrophotometric method for the determination of pectin methylesterase activity of besthorns hydrazone Pifferi, P. G.;A. Malacarne;G. Lanzarini;U. Casoli
  13. Carbohydr. Polyme v.38 Quantitative determination of chitosan by ninhydrin Sabina, P.;M. Kjill;O. Kjetill