Kinetic and Thermodynamic Properties of Partially Purified Dextranase from Paecilomyces lilacinus and Its Application in Dextran Removal from Cane Juice

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• 作者：Surekha Bhatia ; Gaurav Bhakri ; Maninder Arora ; S. K. Batta ; S. K. Uppal
• 关键词：Dextran ; Dextranase ; Paecilomyces lilacinus ; Purification ; Sugar industry ; Sugarcane juice ; Thermodynamic properties
• 刊名：Sugar Tech
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：18
• 期：2
• 页码：204-213
• 全文大小：877 KB
• 参考文献：Abdel-Aziz, M.S., Talkhan N. Fatma, and J.C. Janson. 2007. Purification and characterization of dextranase from a new strain of Penicillium funiculosum. Journal of Applied Sciences Research 3: 1509–1516.
  Arnold, W.N., T.B.P. Nguyen, and L.C. Mann. 1998. Purification and characterization of a dextranase from Sporothrix schenckii. Archives of Microbiology 170: 91–98.CrossRef PubMed
  Bhatia, S., Jyoti, S.K. Uppal, K.S. Thind, and S.K. Batta. 2009. Post harvest quality deterioration in sugarcane under different environmental conditions. Sugar Tech 11: 154–160.CrossRef
  Bhatia, S., Jyoti, S.K. Uppal, K.S. Thind, and S.K. Batta. 2012. Partial purification and characterization of acid invertase from the fresh and stale sugarcane juice. Sugar Tech 14: 148–155. doi:10.​1007/​s12355-012-0137-1 .
  Chen, J.C.P. 1993. Cane sugar hand book. New York: Wiley.
  Chen, L., X. Zhou, W. Fan, and Y. Zhang. 2008. Expression, purification and characterization of a recombinant Lipomyces starkeyi dextranase in Pischia pastoris. Protein Expression and Purification 58: 87–93.CrossRef PubMed
  Clarke, M.A., E.J. Roberts, M.A. Goldshell, M.A. Brannan, F.G. Carpenter, and E.E. Coll. 1980. Sucrose loss in manufacture of cane sugar. Sugar Y Azucar 75: 64–68.
  Dixon, M. 1953. The effect of pH on the affinity of enzymes for substrates and inhibitors. Biochemical Journal 55: 161–171.CrossRef PubMed PubMedCentral
  Dixon, M., and E.C. Webb. 1964. Enzymes, 2nd ed, 84–87. London: Longman Green.
  Egan, B.T. 1967. Gum content and pH as measures of the losses due to sour storage rot. Proceedings of Queensland Society of Sugarcane Technology 35: 31–37.
  Eggleston, G., A. Monge, B. Montes, and D. Steward. 2009. Application of dextranases in sugarcane factory: Overcoming practical problems. Sugar Tech 11: 135–141.CrossRef
  Galvez-Mariscal, A., and A. Lopez-Mungia. 1991. Production and characterization of a dextranase from an isolated Paecilomyces lilacinus strain. Applied Microbiology and Biotechnology 36: 327–331.CrossRef
  Hattori, A., K. Ishibashi, and S. Minato. 1981. The purification and characterization of dextranase of Chaetomium gracile. Agricultural and Biological Chemistry 45: 2409–2416.CrossRef
  Igarashi, T., and A. Yamamoto. 1988. Purification and characterization of intracellular dextranase of Bacteroides oralis Ig4a. Showa Shigakkai Zasshi 8: 405–412.PubMed
  Igarashi, T., H. Morisaki, and N. Goto. 2004. Molecular characterization of dextranase from Sptreptococcus rattus. Microbiology and Immunology 48: 155–162.CrossRef PubMed
  Igarashi, T., A. Yamamoto, and N. Goto. 1992. Characterization of the dextranase purified from Streptococcus mutans Ingbritt. Microbiology and Immunology 36: 969–976.CrossRef PubMed
  Imrie, F.K.E., and R.H. Tilbury. 1972. Polysaccharides in sugarcane and its products. Sugar Tech 1: 291–361.
  Jimenez, E.R. 2005. The dextranase along sugar-making industry. Biotechnologia Aplicada 22: 20–27.
  Jimenez, E.R. 2009. Dextranase in sugar industry: A review. Sugar Tech 11: 124–134.CrossRef
  Khalikova, E., P. Susi, N. Usanov, and T. Korpela. 2003. Purification and properties of extracellular dextranase from a Bacillus sp. Journal of Chromatography B 796: 315–326.CrossRef
  Kobayashi, M., S. Takagi, M. Shiota, Y. Mitsuishi, and K. Matsuda. 1983. An isomaltotriose-producing dextranase from Flavobacterium sp M-73: Purification and properties. Agricultural and Biological Chemistry 47: 2585–2593.CrossRef
  Kobayashi, M., I. Yokoyama, and K. Matsuda. 1984. Substrate binding sites of Leuconostoc dextransucrase evaluated by inhibition kinetics. Agricultural and Biological Chemistry 50: 2585–2590.CrossRef
  Koenig, D., and D. Day. 1989. The purification and characterization of a dextranase from Lipomyces starkeyi. European Journal of Biochemistry 183: 161–167.CrossRef PubMed
  Lee, J.M., and P.F. Fox. 1985. Purification and characterization of Paecilomyces lilacinus dextranase. Enyzme and Microbial Technology 7: 573–577.CrossRef
  Lineweaver, H., and D. Burk. 1934. Determination of enzyme dissociation constants. Journal of the American Chemical Society 56: 658–666.CrossRef
  Lowry, O.H., N.J. Rosebrough, A.L. Farr, and R.J. Randall. 1951. Protein measurement with Folin phenol reagent. Journal of Biological Chemistry 193: 265–275.PubMed
  Madhu, Shukla G.L., and K.A. Prabhu. 1984. Application of dextranase in the removal of dextran from cane juice. International Sugar Journal 86: 136–138.
  Mandel, M., R. Anreotti, and C. Roche. 1976. Measurement of saccharifying cellulose. Biotechnology and Bioengineering Symposium 6: 21–23.
  Morel du Boil, P.G., and S. Wienese. 2000. Enzymatic reduction of dextran in process. Laboratory evaluation of dextranases. Proceedings of South African Sugar Tech Association 76: 435–443.
  Nelson, N. 1944. A photometric adaptation of the Somogyi method for the determination of glucose. Journal of Biological Chemistry 153: 375–380.
  Okada, G., T. Takayanagi, and T. Sawai. 1988a. Improved purification and further characterization of an Isomalto dextranase from Arthrobacter globiformis T 6 . Agricultural and Biological Chemistry 52: 495–501.CrossRef
  Okada, G., T. Unno, and T. Sawai. 1988b. A simple purification procedure for and further characterization of a glucodextranase from Arthrobacter globiformis 142. Agricultural and Biological Chemistry 52: 2169–2176.CrossRef
  Pleszczynska, M., J. Rogalski, J. Szczodrak, and J. Fiedurek. 1996. Purification and some properties of an extracellular dextranse from Penicillium notatum. Mycological Research 100: 681–686.CrossRef
  Pleszczynska, M., J. Szczodrak, J. Rogalski, and J. Fiedurek. 1997. Hydrolysis of dextran by Penicllium notatum dextranase and identification of final digeston products. Mycological Research 101: 69–72.CrossRef
  Rauh, J.S., J.A. Cuddihy, and R.N. Falgout. 2001. Analyzing dextran in sugar industry. A review of dextran in factory and a new analytical technique. Proceedings of the West Indies Sugar Tech Conference 27: 1–12.
  Roberts, E.J., M.A. Clarke, and A.N. Goldshall. 1983. The analysis of dextran in sugar production. Proceedings of the Inernational Society of Sugar Cane Technol 18th Congress 3: 1374–1383.
  Sharma, K.P., B.K. Rattan, and R.S. Kanwar. 1989. Improvement in sugar recovery by preventing bacterial polysaccharide production in cane juice. Bharatiya Sugar 14: 15–18.
  Shimizu, E., T. Unno, M. Ohba, and G. Okoda. 1998. Purification and characterization of an Isomaltotriose-producing endo-dextranase from Fusarium sp. Bioscience, Biotechnology, and Biochemistry 62: 117–122.CrossRef PubMed
  Simonson, L.G., A.E. Liberta, and A. Richardson. 1975. Characterization of an extracellular dextranase from Fusarium monoliforme. Applied Microbiology 30: 855–861.PubMed PubMedCentral
  Solomon, S. 2009. Post harvest deterioration of sugarcane. Sugar Tech 11: 109–123.CrossRef
  Solomon, S., P. Singh, A.K. Shrivastava, P. Singh, A. Chandra, R. Jani, and C.P. Prajapati. 2011. Physico-chemical method of preserving sucrose in harvested sugarcane at high ambient temperature in a Sub tropical climate. Sugar Tech 13: 60–67.CrossRef
  Sugiura, M., A. Ito, T. Ogiso, K. Kato, and H. Asano. 1973. Purification of dextranase from Penicilluim funiculosum and its enzymatic properties. Biochimica et Biophysica Acta (BBA)—Enzymol 309: 357–362.CrossRef
  Sun, J., X. Cheng, Y. Zhang, Z. Yan, and S. Zhang. 1988. A strain of Paecilomyces lilacinus producing high quality dextranases. Purification and characterization. Annals of the New York Academy of Sciences 542: 192–194.CrossRef
  Webb, E., and I. Spencer-Martins. 1983. Extracellular endodextranase from the yeast Lipomyces starkeyi. Canadian Journal of Microbiology 29: 1092–1095.CrossRef
  Wynter, C., B.K. Patel, P. Bain, J. De Jersey, S. Hamilton, and P.A. Inkerman. 1996. A novel thermostable dextranase from a Thermoanaerobacter species cultured from the geothermal waters of the Great Artesian Basin of Australia. Microbiology Letters 140: 271–276.CrossRef
  
• 作者单位：Surekha Bhatia (1)
  Gaurav Bhakri (2)
  Maninder Arora (3)
  S. K. Batta (2)
  S. K. Uppal (4)
  
  1. Department of Processing and Food Engineering, Punjab Agricultural University, Ludhiana, 141004, India
  2. Department of Biochemistry, Punjab Agricultural University, Ludhiana, 141004, India
  3. Department of Microbiology, Punjab Agricultural University, Ludhiana, 141004, India
  4. Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana, 141004, India
  
• 刊物主题：Agriculture;
• 出版者：Springer India
• ISSN：0974-0740

文摘

Dextran is a high molecular weight polysaccharide formed in sugarcane during post harvest staling resulting in loss of sucrose as well as quality of sugar. Information regarding the removal of polysaccharides from the sugarcane milled juice is limited. In the present study, enzyme dextranase was produced from Paecilomyces lilacinus by submerged fermentation using Mandel medium and substrate dextran. The crude enzyme was partially purified by 80 % ammonium sulphate saturation followed by DEAE-cellulose column chromatography. Two isoforms of dextranase D-I and D-II each with optimum pH 5.0 and temperature 50 °C were identified. Both the isoforms were most efficient kinetically at pH 5.0 and temperature 50 °C and were found to be thermostable in the temperature range 0–50 °C. The pKa values of ionizing groups in free enzyme and enzyme-substrate complex were found to be between 4.1 and 5.8 indicating the possible participation of carboxylate groups of aspartate/ glutamate and imidazolium group of histidine in dextranase catalysed hydrolysis of dextran by both the isoforms. Activation energy (Ea) values for D-I and D-II were 22.07 and 31.68 kJ/mol and corresponding enthalpy change (ΔH) values were 17.01 and 8.70 kJ/mol, respectively. Cu2+ activated whereas Mg2+, Ca2+, Mn2+, Fe2+ and Pb2+ inhibited the activity of dextranase. With the application of 5, 10 and 15 units of partially purified dextranase per 100 mL of juice, dextran content decreased by 56.39, 73.88 and 80.27 %, respectively as compared to the control (with no dextranase added) after 24 h of storage of cane juice.