1887

Abstract

An extracellular tannase was produced from solid-state cultures of . The enzyme was purified to homogeneity from the cell-free culture broth by preparative isoelectric focusing and by FPLC using anion-exchange and gel-filtration chromatography. SDS-PAGE analysis as well as gel localization studies of purified tannase indicated the presence of two enzyme forms, with molecular masses of 90 kDa and 180 kDa. The tannase had an isoelectric point of 3·8, a temperature optimum of 60–70 °C and a pH optimum of 6·0. The substrate specificity of the tannase was determined by HPLC analysis of tannin substrates and products. The enzyme was able to remove gallic acid from both condensed and hydrolysable tannins. Internal sequences were obtained from each of the gel-purified and trypsin-digested tannase forms. The peptide sequences obtained from both forms were identical to sequences within a -glucosidase from . The purified tannase was tested for -glucosidase activity and was shown to hydrolyse cellobiose efficiently. However, no -glucosidase activity was detected when the enzyme was assayed in the presence of tannic acid.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26346-0
2003-10-01
2019-11-12
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/10/mic1492941.html?itemId=/content/journal/micro/10.1099/mic.0.26346-0&mimeType=html&fmt=ahah

References

  1. Adachi, O., Watanabe, M. & Yamada, H. ( 1968; ). Studies on fungal tannase. Part II. Physicochemical properties of tannase of Aspergillus flavus. Agric Biol Chem 32, 1079–1085.[CrossRef]
    [Google Scholar]
  2. Agbo, F. & Spradlin, J. E. ( 1995; ). Enzymatic clarification of tea extracts. US Patent no. 5 445 836.
  3. Aguilar, C. N., Augur, C., Favela-Torres, E. & Viniegra-González, G. ( 2001; ). Induction and repression patterns of fungal tannase in solid-state and submerged cultures. Process Biochem Int 36, 565–570.[CrossRef]
    [Google Scholar]
  4. Aoki, K., Shinke, R. & Nishira, H. ( 1976; ). Chemical composition and molecular weight of yeast tannase. Agric Biol Chem 40, 297–302.[CrossRef]
    [Google Scholar]
  5. Aoki, K., Tanaka, T., Shinke, R. & Nishira, H. ( 1979; ). Detection of tannase in polyacrylamide gels. J Chromatogr 170, 446–448.[CrossRef]
    [Google Scholar]
  6. Barmentlo, B., Bel, W. J., Hoogstad, B., Pendlington, S. & Slater, N. K. ( 1993; ). Process of preparing a tea product. US Patent no. 5 258 188.
  7. Barthomeuf, C., Regerat, F. & Pourrat, H. ( 1994; ). Production, purification and characterization of a tannase from Aspergillus niger LCF 8. J Ferment Bioeng 77, 320–323.[CrossRef]
    [Google Scholar]
  8. Beverini, M. & Metche, M. ( 1990; ). Identification, purification and physicochemical properties of tannase of Aspergillus oryzae. Sci Aliment 10, 807–816.
    [Google Scholar]
  9. Boadi, D. K. & Neufeld, R. J. ( 2001; ). Encapsulation of tannase for the hydrolysis of tea tannins. Enzyme Microb Technol 28, 590–595.[CrossRef]
    [Google Scholar]
  10. Chittum, H. S., Lane, W. S., Carlson, B. A., Roller, P. P., Lung, F. D. T., Byeong, J. L. & Hatfield, D. L. ( 1998; ). Rabbit β-globin is extended beyond its UGA stop codon by multiple suppression and translational reading gaps. Biochemistry 37, 10866–10870.[CrossRef]
    [Google Scholar]
  11. Deschamps, A. M., Mahodeau, G., Conti, M. & Lebeault, J. M. ( 1980; ). Bacteria degrading tannic acid and related compounds. J Ferment Technol 58, 93–97.
    [Google Scholar]
  12. Díaz-Godínez, G., Soriano-Santos, J., Augur, C. & Viniegra-González, G. ( 2001; ). Exopectinases produced by Aspergillus niger in solid-state fermentation and submerged fermentation: a comparative study. J Ind Microbiol Biotechnol 26, 271–275.[CrossRef]
    [Google Scholar]
  13. Farias, G. M., Gorbea, C., Elkins, J. R. & Griffin, G. J. ( 1994; ). Purification, characterization and substrate relationships of the tannase from Chryphonectria parasitica. Physiol Mol Plant Pathol 44, 51–63.[CrossRef]
    [Google Scholar]
  14. Garcia-Peña, I., Favela-Torres, E. & Augur, C. ( 1999; ). Purificación parcial de tanasa producida por Aspergillus niger en fermentación en medio sólido. In Topicos in Biotecnología, Avances en Purificación y Aplicación de Enzimas en Biotecnología, pp. 247–261. Edited by A. Prado-Barragan, S. Huerta-Ochoa, G. Rodriguez-Serrano & G. Saucedo Castañeda. Mexico City: UAM-I.
  15. Hatamoto, O., Watarai, T., Kikuchi, M., Misusawa, K. & Sekine, H. ( 1996; ). Cloning and sequencing of the gene encoding tannase and structural study of the tannase subunit from Aspergillus oryzae. Gene 175, 215–221.[CrossRef]
    [Google Scholar]
  16. Hegyi, H. & Gernstein, M. ( 1999; ). The relationship between protein structure and function: a comprehensive survey with application to the yeast genome. J Mol Biol 288, 147–164.[CrossRef]
    [Google Scholar]
  17. Iwashita, K., Todoroki, K., Kimura, H., Shimoi, H. & Ito, K. ( 1998; ). Purification and characterization of extracellular and cell wall bound β-glucosidases from Aspergillus kawachii. Biosci Biotechnol Biochem 62, 1938–1946.[CrossRef]
    [Google Scholar]
  18. Iwashita, K., Nagahara, T., Kimura, H., Takano, M., Shimoi, H. & Ito, K. ( 1999; ). The bglA gene of Aspergillus kawachii encodes both extracellular and cell wall-bound beta-glucosidases. Appl Environ Microbiol 65, 5546–5553.
    [Google Scholar]
  19. Kilkowski, W. J. & Gross, G. G. ( 1999; ). Color reaction of hydrolyzable tannins with Bradford reagent Coomassie brilliant blue. Phytochemistry 51, 363–366.[CrossRef]
    [Google Scholar]
  20. Kumar, R. A., Gunasekaran, P. & Lakshmanan, M. ( 1999; ). Biodegradation of tannic acid by Citrobacter freundii isolated from a tannery effluent. J Basic Microbiol 39, 161–168.[CrossRef]
    [Google Scholar]
  21. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  22. Lekha, P. K. & Lonsane, B. K. ( 1994; ). Comparative titers, location and properties of tannin acyl hydrolase produced by Aspergillus niger PKL 104 in solid-state, liquid surface and submerged fermentations. Process Biochem 29, 497–503.[CrossRef]
    [Google Scholar]
  23. Mondal, K. C. & Pati, B. R. ( 2000; ). Studies on the extracellular tannase from newly isolated Bacillus licheniformis KBR 6. J Basic Microbiol 40, 223–232.[CrossRef]
    [Google Scholar]
  24. Niehaus, J. U. & Gross, G. G. ( 1997; ). A gallotannin degrading esterase from leaves of pedunculate oak. Phytochemistry 45, 1555–1560.[CrossRef]
    [Google Scholar]
  25. Ramírez-Coronel, M. A. ( 2000; ). Purificación y caracterización de una tanasa de A. niger AA20 producida por fermentacion en medio sólido. Master's thesis, Universidad Autónoma Metropólitana Iztapalapa, Mexico.
  26. Romero-Gómez, S., Augur, C. & Viniegra-González, G. ( 2000; ). Invertase production by Aspergillus niger in submerged and solid state fermentation. Biotechnol Lett 22, 1255–1258.[CrossRef]
    [Google Scholar]
  27. Sang, J. H., Yong, J. Y. & Hyen Sam, K. ( 1995; ). Characterization of a bifunctional cellulase and its structural gene. The cel gene of Bacillus sp. D04 has exo and endoglucanase activity. J Biol Chem 270, 26012–26019.[CrossRef]
    [Google Scholar]
  28. Seth, M. & Chand, S. ( 2000; ). Biosynthesis of tannase and hydrolysis of tannins to gallic acid by Aspergillus awamori, optimization of process parameters. Process Biochem 36, 39–44.[CrossRef]
    [Google Scholar]
  29. Suseela, R. G. & Nandy, S. C. ( 1983; ). Isolation, purification and some properties of Penicillium Chrysogenum tannase. Appl Environ Microbiol 46, 525–527.
    [Google Scholar]
  30. Yamada, D. & Tanaka, T. ( 1972; ). Wine making using tannase in fermentation process. Patent. Ger. Offen. 2.224 100 (cl. 12g). Japanese Patent Application no. 71 33, 151.
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26346-0
Loading
/content/journal/micro/10.1099/mic.0.26346-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error