Skip to main content
SpringerLink
Log in

Purification and Characterization of Pectin Lyase Produced by Aspergillus terricola and its Application in Retting of Natural Fibers

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

An indigenously isolated fungal strain identified as Aspergillus terricola with assigned fungal strain number MTCC 7588 has been used as source for pectin lyase production. The extracellular pectin lyase was purified to homogeneity from the culture filtrate of A. terricola by ion exchange and gel filtration chromatography. The determined molecular weight was 35 ± 01 kDa. The K m and k cat (turnover) values of the purified enzyme at 37 °C using citrus pectin as the substrate were found to be 1.0 mg/ml and 110.0 s−1, respectively. The pH and temperature optima of the enzyme were 8.0 and 50 °C, respectively. The retting ability of the purified pectin lyase for natural fibers viz. Cannabis sativa and Linum usitatissimum has been demonstrated for the first time.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Schols, H. A., Geraeds, C. C. J. M., Searle-van Leeuwen, M. F., Kormelink, F. J. M., & Voragen, A. G. J. (1990). Rhamnogalacturonase: A novel enzyme that degrades the hairy regions of pectins. Carbohydrate Research, 206, 106–115.

    Google Scholar 

  2. Suykerbuyk, M. E. G., Schaap, P. J., Musters, W., & Visser, J. (1995). Cloning, sequence and expression of the gene for rhamnogalacturonan hydrolase of Aspergillus aculeatus; a novel pectinolytic enzyme. Applied Microbiology and Biotechnology, 43, 861–870. doi:10.1007/BF02431920.

    Article  CAS  Google Scholar 

  3. Kofod, L. V., Kauppinen, S., Christgau, S., Andersen, L. N., Heldt-Hansen, H. P., Dorreich, K., et al. (1994). Cloning and characterization of two structurally and functionally divergent rhamnogalacturonases from Aspergillus aculeatus. The Journal of Biological Chemistry, 269, 29182–29189.

    CAS  Google Scholar 

  4. Mutter, M., Colquhoun, I. J., Schols, H. A., Beldman, G., & Voragen, A. G. J. (1996). Rhamnogalacturonase B from Aspergillus aculeatus is a rhamnogalacturonan α-L- rhamnopyranosyl-(1-4)-α-d-galactopyranosyluronide lyase. Plant Physiology, 110, 73–77. doi:10.1104/pp.110.1.73.

    Article  CAS  Google Scholar 

  5. de Vries, R. (1999). Accessory enzymes from Aspergillus involved in xylan and pectin degradation. Netherlands: The Wageningen Agricultural University; PhD. Thesis.

  6. Alkorta, I., Garbisu, C., Liama, M. J., & Serra, J. L. (1998). Industrial application of pectic enzymes: A review. Process Biochem, 33, 21–28. doi:10.1016/S0032-9592(97)00046-0.

    Article  CAS  Google Scholar 

  7. Blanco, P., Sieiro, C., & Villa, T. G. (1999). Production of pectic enzymes in yeast. FEMS Microbiology Letters, 175, 1–9. doi:10.1111/j.1574-6968.1999.tb13595.x.

    Article  CAS  Google Scholar 

  8. Hoondal, G. S., Tewari, R. P., Tewari, R., Dahiya, N., & Beg, Q. K. (2002). Microbial alkaline pectinases and their industrial applications: A review. Applied Microbiology and Biotechnology, 59, 409–418. doi:10.1007/s00253-002-1061-1.

    Article  CAS  Google Scholar 

  9. Delgado, L., Blanco, A. T., Huitron, C., & Aguilar, G. (1992). Pectin lyase from Aspergillus sp. CH-Y-1043. Applied Microbiology and Biotechnology, 39, 515–519. doi:10.1007/BF00205043.

    Article  Google Scholar 

  10. Targano, V. M., & Pelosof, A. M. R. (1994). Application of Doehlert design for water activity, pH and fermentation time, optimization for Aspergillus niger pectinolytic activity and production in solid state and submerged fermentations. Enzyme and Microbial Technology, 25, 411–419. doi:10.1016/S0141-0229(99)00065-4.

    Article  Google Scholar 

  11. Yadav, S., Yadav, P. K., Yadav, D., & Yadav, K. D. S. (2008). Pectin lyase: A review. Process Biochem (in press). doi:10.1016/j.procbio.2008.09.012.

  12. Mollet, J. C., Park, S. Y., Nothnagel, E. A., & Lord, E. M. (2000). A lily stylar pectin is necessary for pollen tube adhesion to an in vitro stylar matrix. The Plant Cell, 12, 1737–1749.

    Article  CAS  Google Scholar 

  13. Sharma, H. S. S. (1988). Chemical retting of flax using chelating compounds. Applied Biology, 113, 159–165. doi:10.1111/j.1744-7348.1988.tb03292.x.

    Article  CAS  Google Scholar 

  14. Van Sumere, C. F. (1992). Retting of flax with special reference to enzyme retting. In H. S. S. Sharma, & C. F. Van Sumere (Eds.), The biology and processing of Flax (pp. 157–198). Belfast Northern Ireland UK: M Publications.

    Google Scholar 

  15. Petrich-Murray, H., Fourtot-Brun, C., Chaereau, D., Decock, B., & Lefevre, J. (1995). Procede de rouissage enzymatique. French patent, 2713671.

  16. Akin, D. E., Henriksson, G., Slomezynski, D., & Eriksson, K. E. L. (1998). Rhizomucor pusillus: Pectinolytic fungus with industrial application for retting flax. Abstracts of the 98th General Meeting of the American Society for Microbiology, 5, 394.

  17. Henriksson, G., Akin, D. E., Slomezynski, D., & Eriksson, K. E. L. (1999). Production of highly efficient enzymes for flax retting by Rhizomucor pusillus. Journal of Biotechnology, 68, 115–123. doi:10.1016/S0168-1656(98)00192-8.

    Article  CAS  Google Scholar 

  18. Akin, D. E., Rigsby, L. L., Perkins, W., Henriksson, G., & Eriksson, K. E. L. (2000). Spray enzymatic retting: A new method for processing flax fibers. Textile Research, 70, 486–494. doi:10.1177/004051750007000604.

    Article  CAS  Google Scholar 

  19. Zhang, J., Henriksson, G., & Johansson, G. (2000). Polygalacturonase is the key enzyme component in enzyme retting of flax. Journal of Biotechnology, 81, 85–89. doi:10.1016/S0168-1656(00)00286-8.

    Article  CAS  Google Scholar 

  20. Akin, D. E., Dodd, R. B., Sohn, M., & Foulk, J. A. (2006). Retting of linseed straw with pectate lyase for flax fiber. Proceeding of 61st flax institution of the United States. pp. 129–135.

  21. Yadav, S., Yadav, P. K., Yadav, D., & Yadav, K. D. S. (2008). Purification and characterization of an alkaline pectin lyase from Aspergillus flavus. Process Biochemistry, 43, 547–552. doi:10.1016/j.procbio.2008.01.015.

    Article  CAS  Google Scholar 

  22. Cappuccino, J. G., & Sherman, N. Microbiology: A laboratory manual, 5th edn (pp. 14–19). California: Benjamin/Cumming.

  23. Molina, S. M. G., Pelissari, F. A., & Vitorello, C. B. M. (2001). Screening and genetic improvement of pectinolytic fungi for degumming of textile fibres. Brazilian Journal of Microbiology, 32, 320–326. doi:10.1590/S1517-83822001000400013.

    Article  Google Scholar 

  24. Albersheim, P. (1966). Pectin Lyase from Fungi. Methods in Enzymology, 8, 628–631. doi:10.1016/0076-6879(66)08113-8.

    Article  CAS  Google Scholar 

  25. Lowry, O. H., Rose Brough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with Folin phenol reagent. The Journal of Biological Chemistry, 193, 265–275.

    CAS  Google Scholar 

  26. Werber, K., & Osborn, M. (1969). The relative mobility or molecular weight determination by dodecyl sulfate-polyacrylamide gel electrophoresis. The Journal of Biological Chemistry, 244, 4406–4412.

    Google Scholar 

  27. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685. doi:10.1038/227680a0.

    Article  CAS  Google Scholar 

  28. Ortega, N., de Diego, S., Rodriguez-Nogales, J. M., Perez-Mateos, M., & Busto, M. D. (2004). Kinetic behaviour and thermal inactivation of pectin lyase used in food processing. International Journal of Food Science & Technology, 39, 631–639. doi:10.1111/j.1365-2621.2004.00822.x.

    Article  CAS  Google Scholar 

  29. Zhang, J., Henriksson, H., Szabo, I. J., Henriksson, G., & Johansson, G. (2005). The active component in the flax retting system of the zygomycetes Rhizopus oryzae sb. is a family 28 polygalacturaonase. Journal of Industrial Microbiology & Biotechnology, 32, 431–443. doi:10.1007/s10295-005-0014-y.

    Article  CAS  Google Scholar 

  30. Zhang, J. (2006). Biochemical study and technical applications of fungal pectinase, Acta Universitatis Upsaliensis. Digital comprehensive summaries of Uppsala Dissertations From Faculty of Science and Technology, 14.

  31. Alana, A., Maria, J. L., & Serra, J. L. (1991). Purification and some properties of pectin lyase from Penicillium italicum. FEBS, 280, 335–340. doi:10.1016/0014-5793(91)80325-W.

    Article  CAS  Google Scholar 

  32. Silva, D. O., Attwod, M. M., & Tempest, D. W. (1993). Partial purification and properties of pectin lyase from Penicillium expansum. World Journal of Microbiology & Biotechnology, 9, 574–578. doi:10.1007/BF00386297.

    Article  CAS  Google Scholar 

  33. Chen, W. C., Hsieh, H. J., & Tseng, T. C. (1998). Purification and characterization of a pectin lyase from Pythium splendens infected cucumber fruits. Botanical Bulletin of Academia Sinica, 38, 181–186.

    Google Scholar 

  34. Adejuwon, A. O., & Olutiola, P. O. (2007). Pectin lyase activity in culture medium of Lasidioplodia theobromae. Journal of Plant Sciences, 2, 334–340.

    Article  CAS  Google Scholar 

  35. Yadav, S., Yadav, P. K., Yadav, D., & Yadav, K. D. S. (2008). Purification and characterization of an acidic pectin lyase produced by Aspergillus ficuum strain MTCC 7591 suitable for clarification of fruit juices. Annals of Microbiology, 58, 61–65.

    Article  CAS  Google Scholar 

  36. Spagana, G., Pifferi, P. G., & Gilioli, E. (1995). Immobilization of a pectin lyase from Aspergillus niger for application in food technology. Enzyme and Microbial Technology, 17, 729–738. doi:10.1016/0141-0229(94)01134-I.

    Article  Google Scholar 

  37. Hamdy, H. S. (2005). Purification and characterization of the pectin lyase produced by Rhizopus oryzae grown on orange peels. Annals of Microbiology, 55, 205–211.

    CAS  Google Scholar 

  38. Sinitsyna, O. A., Fedorova, E. A., Semenova, M. V., Gusakov, A. V., Sokolova, L. M., & Bubnova, T. M. (2007). Isolation and characterization of extracellular pectin lyase from Penicillium canescens. Biochemist (Moscow), 72, 565–567. doi:10.1134/S0006297907050148.

    Article  CAS  Google Scholar 

  39. Dixon, M., & Webb, E. C. (1979). Enzyme kinetics. In Enzyme, 3rd ed. (pp. 47). New York, San Francesco: Academic Press.

  40. Szajer, I., & Szajer, C. (1982). Pectin lyase of Penicillium paxilli. Biotechnology Letters, 4, 549–552. doi:10.1007/BF00127782.

    Article  CAS  Google Scholar 

  41. Ishii, S., & Yokotsuka, T. (1972). Purification and properties of pectin trans-eliminase from Aspergillus sojae. Agricultural and Biological Chemistry, 36, 146–153.

    CAS  Google Scholar 

  42. Ishii, S., & Yokotsuka, T. (1975). Purification and properties of pectin lyase from Aspergillus japonicus. Agricultural and Biological Chemistry, 39, 313–321.

    CAS  Google Scholar 

  43. Bugbee, W. M. (1990). Purification and characteristics of pectin lyase from Rhizoctonia solani. Physiological and Molecular Plant Pathology, 36, 15–25. doi:10.1016/0885-5765(90)90088-F.

    Article  CAS  Google Scholar 

  44. Yadav, S., & Shastri, N. V. (2007). Purification and properties of an extracellular pectin lyase produced by the strain of Penicillium oxalicum in solid-state fermentation. Indian Journal of Biochemistry & Biophysics, 44, 247–251.

    CAS  Google Scholar 

  45. Guevara, M. A., Gonzalez-Jaen, M. T., & Estevez, P. (1996). Pectin lyase from Fusarium oxysporum f. sp. radicis lycopersici: purification and characterization. Progress in Biotechnology, 14, 747–760. doi:10.1016/S0921-0423(96)80312-1.

    Article  CAS  Google Scholar 

  46. Lim, J. Y., Fujio, Y., & Ueda, S. (1983). Purification and characterization of pectinesterase and pectin lyase from Aspergillus oryzae A-3. Journal of Applied Biochemistry, 5, 91–98.

    CAS  Google Scholar 

  47. Mayans, O., Scott, M., Connerton, I., Gravesen, T., Benen, J., & Visser, J. (1997). Two crystal structures of pectin lyase A from Aspergillus reveal a pH driven conformational change and striking divergence in the substrate-binding clefts of pectin and pectate lyases. Structure (London, England), 5, 677–689. doi:10.1016/S0969-2126(97)00222-0.

    CAS  Google Scholar 

  48. Linen, H. I. T. (1986). Textiles, 15, 30–34.

    Google Scholar 

  49. Schunke, H., Sanio, C., Pape, H., Schunke, U., & Matz, C. (1995). Reduction of time required for dew retting of flax: influence of agricultural, mechanical and microbiological techniques on fiber processing. Melliand, 76, E101–E104.

    Google Scholar 

  50. Adamsen, A. P. S. (2002). Chelating agents and enzyme retting of flax. Textile Research Journal, 72, 296–302. doi:10.1177/004051750207200404.

    Article  CAS  Google Scholar 

  51. Henriksson, G., Akin, D. E., Rigsby, L. L., Patel, N., & Eriksson, K. E. L. (1997). Influence of chelating agents and mechanical pretreatment on enzymatic retting of flax. Textile Research Journal, 67, 829–836.

    CAS  Google Scholar 

  52. Jacob, N., & Prema, P. (2008). Novel process for the simultaneous extraction and degumming of banana fibers under solid-state cultivation. Brazilian Journal of Microbiology, 39, 115–121. doi:10.1590/S1517-83822008000100025.

    Article  Google Scholar 

Download references

Acknowledgements

Sangeeta Yadav is thankful to the Department of Science and Technology, Government of India, New Delhi for the financial support in the form of a Women Scientist Fellowship SR/WOS-A/LS-34/2004. The laboratory facilities provided by the Heads of the Chemistry Department and Biotechnology Department, D.D.U. Gorakhpur University, Gorakhpur are also thankfully acknowledged.

Author information

Authors and Affiliations

  1. Department of Chemistry, Deen Dayal Upadhyaya Gorakhpur University, Gorakhpur, Uttar Pradesh, 273 009, India

    Sangeeta Yadav & Kapil Deo Singh Yadav

  2. Department of Biotechnology, D.D.U. Gorakhpur University, Gorakhpur, Uttar Pradesh, 273009, India

    Pramod Kumar Yadav & Dinesh Yadav

  3. Department of Molecular Biology and Genetic Engineering, CBSH, G.B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, 263 145, India

    Dinesh Yadav

Authors
  1. Sangeeta Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pramod Kumar Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dinesh Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kapil Deo Singh Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sangeeta Yadav.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yadav, S., Yadav, P.K., Yadav, D. et al. Purification and Characterization of Pectin Lyase Produced by Aspergillus terricola and its Application in Retting of Natural Fibers. Appl Biochem Biotechnol 159, 270–283 (2009). https://doi.org/10.1007/s12010-008-8471-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-008-8471-1

Keywords

Search

Navigation