Laundry Detergent Compatibility of Papain Like Protease Purified From Piper Betel Leaves

  • Abstract
  • Keywords
  • References
  • PDF
  • Abstract

    Enzymes have wide applications in detergent industry from early 1900’s. Mostly, clothes are soiled by protein based grime. Most of the detergents have either amylase / protease. Various sources were scrutinized for potent protease activity and Betel leaves were selected, the enzyme was then isolated, purified to homogeneity by ammonium sulphate precipitation, DEAE-Cellulose and gel permeation chromatographic techniques. The enzyme was monomeric in nature with a molecular mass of 38kDa as determined by native PAGE and SDS-PAGE. The enzyme shows maximum activity at 60oC and pH 4.0. The Km and Vmax of the enzyme were 4x10-3M and 54µmol/min/mg respectively. The enzyme was categorically inhibited by PCMB and iodo-acetamide suggesting it to have papain like nature. The stability of the enzyme is assessed over the stretch of alkaline pH and temperature. This evaluation validates the stability of the enzyme and its use in detergent formulations. It was evident that after adding the enzyme preparation the stains (tea, chocolate, blood) were removed much better than that of the controls, which affirms that papain like enzyme from betel leaves, enhances detergent activity.



  • Keywords

    Betel Leaves; Detergent Activity ; Enzyme Kinetics; Purification.

  • References

      [1] Siezen, R. J., & Leunissen, J. A., “Subtilases: the superfamily of subtilisin‐like serine proteases”, Protein Science, 6(3), (1997), 501-523.

      [2] Duffy, J. I., & Gutcho, S., “Chemicals by enzymatic and microbial processes: recent advances [USA]”, Noyes Data Corp, (1980)

      [3] Cohen, L. W., Coghlan, V. M., & Dihel, L. C., “Cloning and sequencing of papain-encoding cDNA”, Gene, 48(2), (1986), 219-227.

      [4] Drenth, J., J. N. Jansonius, R. Koekoek, H. M. Swen, and B. G. Wolthers, “Structure of papain” Nature, 218, (1968), 929-932.

      [5] Drenth, J., Kalk, K. H., & Swen, H. M., “Binding of chloromethyl ketone substrate analogs to crystalline papain”, Biochemistry, 15(17), (1976), 3731-3738.

      [6] Varughese, K. I., Ahmed, F. R., Carey, P. R., Hasnain, S., Huber, C. P., & Storer, A. C., “Crystal structure of a papain-E-64 complex”, Biochemistry, 28(3), (1989), 1330-1332.

      [7] Schechter, I., & Berger, A., “On the size of the active site in proteases. I. Papain”, Biochem. Biophys. Res. Commun, 27, (1967), 157-162.

      [8] Lopes, M. C., Mascarini, R. C., da Silva, B. M. C. G., Flório, F. M., & Basting, R. T., “Effect of a papain-based gel for chemo-mechanical caries removal on dentin shear bond strength”, Journal of Dentistry for Children, 74(2), (2007), 93-97.

      [9] Burrows, D. L., Nicolaides, A., Rice, P. J., Dufforc, M., Johnson, D. A., & Ferslew, K. E., “Papain: a novel urine adulterant”, Journal of analytical toxicology, 29(5), (2005), 275-295.

      [10] Anson, M. L., “The estimation of pepsin, trypsin, papain, and cathepsin with hemoglobin”, The Journal of General Physiology, 22(1) (1938)79-89.

      [11] Liener, I. E., & Kakade, M. L., “Protease inhibitors in" Toxic constituents in plant foodstuffs”, Ed. Liener, IE, (1969), 7.

      [12] Arnon, R., “Papain”, Methods in enzymology, 19, (1970), 226-244.

      [13] Laemmli, U. K., “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”, Nature, 227(5259), (1970), 680-685.

      [14] Dubey, V. K., Pande, M., Singh, B. K., & Jagannadham, M. V., “Papain-like proteases: Applications of their inhibitors”, African Journal of Biotechnology, 6(9), (2007), 1077-1086.

      [15] Adinarayana, K., Raju, K. B., & Ellaiah, P., “Investigations on alkaline protease production with B. subtilis PE-11 immobilized in calcium alginate gel beads”, Process Biochemistry, 39(11), (2004), 1331-1339.

      [16] Wahyuntari, B., Mubarik, N. R., & Anggaran, M., “Isolation and selection of alkaline proteolytic bacteria from leather processing waste and enzyme characterization”, Biotropia-The Southeast Asian Journal of Tropical Biology, 22, (2004).

      [17] Phadatare, S. U., Deshpande, V. V., & Srinivasan, M. C., “High activity alkaline protease from Conidiobolus coronatus (NCL 86.8. 20): enzyme production and compatibility with commercial detergents”, Enzyme and microbial technology, 15(1), (1993), 72-76.




Article ID: 14506
DOI: 10.14419/ijet.v7i3.3.14506

Copyright © 2012-2015 Science Publishing Corporation Inc. All rights reserved.