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A Study on Downstream Processing for the production of Pullulan by Aureobasidium pullulans-SB-01 from the Fermentation broth

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Author Affiliations

  • 1Instrumental and Food Analysis Laboratory, School of Bio Sciences and Technology, VIT University, Vellore, Tamil Nadu, INDIA

Res. J. Recent Sci., Volume 2, Issue (ISC-2012), Pages 16-19, February,2 (2013)

Abstract

Pullulan, which is made up of linear α-D-glucan maltotriose and maltotetrose repeating units interconnected by α (1→6) and α (1→4) linkages, is a water-soluble homopolysaccharide produced extracellularly by Aureobasidium pullulans. Although the production of this bio-polymer is commercially going on still the establishment of the cost effective downstream processing has not attained up to the mark. It is necessary to harvest cells, remove the melanin pigments co-produced during its fermentation followed by its precipitation, concentration and drying. The present work reports on some of these aspects. Centrifugation of the fermentation broth at 8,000 rpm for 20 min gave cell pellets that were discarded and a greenish black supernatant containing melanin pigment which was subjected to the heat treatment at 80°C for 30 min in order to remove the protein (mainly Pullulanase) in the fermentation broth. The supernatant was demelanized by with hydrogen peroxide and activated charcoal, solvent-solvent blends, or by solvent-salt combinations in which hydrogen peroxide treatment shows better result for the removal of melanin pigments. For the precipitation of the exopolysachride the cold Isopropanol was used followed by its drying process at 60°C for 40 min. This methodology produced high purity pullulan that was comparable in colour and texture to the commercial samples which was characterized by the HPLC and FT-IR analysis.

References

  1. Suneetha V., Sindhuja K.V., Sanjeev K., Screening characterization and optimization of Pullulan producing microorganisms from chitoor district, Asian J Microbiol Biotech Env. Sci., 12(2), 149-155 (2010)
  2. Cheng K.C., Demirci A., Catchmark J.M., Evaluation of medium composition and fermentation parameters on pullulan production by Aureobasidium pullulans, Food Sci Tech Int, 17(3), 99-109 (2011)
  3. Cheng K.C., Demirci A., Catchmark J.M. Pullulan: biosynthesis, production, and applications, App Microbiol Biotech, 92(1), 29-44 (2011)
  4. Chi Z.M. and Zhao S.Z. Optimization of medium and cultivation conditions for pullulan production by a new pullulan-producing yeast strain, Enz Microbiol Tech, 33(1), 206–211 (2003)
  5. Li B.X., Zhang N., Peng Q., Yin T., Guan F.F., Wang G.L., Li Y. Production of pigment-free pullulan by swollen cell in Aureobasidium pullulans NG which cell differentiation was affected by pH and nutrition, Appl Microbiol Biotech, 84(2), 293-300 (2009)
  6. Madi N.S., Harvey L.M., Mehlert A., McNeil B. Synthesis of two distinct exopolysaccharide fractions by cultures of the polymorphic fungus Aureobasidium pullulans, Car Polymers 32(3), 307–314 (1997)
  7. Bishwambhar M., Suneetha V. and Kalyani R. The role of microbial pullulan, a biopolymer in pharmaceutical approaches: A review, J App Pharma Sci, 01(06), 45-50 (2011)
  8. Bishwambhar M. and Suneetha V. Characterization of exopolysaccharide a pullulan produced by a novel strain of Aureobasidium pullulans-SB-1 isolated from the phylloplane of Brassica oleracea cultivated in Orissa State, Asian J Microbiol Biotechnol Env Sci , 14 (3),369-374 (2012)
  9. Milanka D.R., Olga G.C., Snezana D.N., Dragana S.D. et al. Simultaneous production of pullulan and biosorption of metals by Aureobasidium pullulans strain CH-1 on peat hydrolysate, Bioresource Tech, 99(4), 6673–6677 (2008)
  10. Pollock J.T., Thorne L. and Armentront R.W., Isolation of new Aureobasidium strains that produce high molecular weight pullulan with reduced pigmentation, Appl Env Microbiol, 5(2), 877–883 (1992)
  11. Singh R.S. and Saini G.K., Pullulan-hyperproducing color variant strain of Aureobasidium pullulans FB-1 newly isolated from phylloplane of Ficus sp. Bio res. Tech, 99(3), 3896–3899 (2008)
  12. Punnapayak H., Sudhadham M., Prasongsuk S., Pichayangkura S. Characterization of Aureobasidium pullulans isolated from airborne spores in Thailand, J. Industrial Microbiol Biotechnol, 30(4), 89–94 (2003)
  13. Hyung-Pil Seo, Chang-Woo Son , Chung-Han Chung , Dae-Il Jung, Sung-Koo Kim, Richard A. Gross, David L Kaplan, Jin-Woo Lee Production of high molecular weight pullulan by Aureobasidium pullulans HP-2001 with soybean pomace as a nitrogen source, Bio res Tech, 95(3), 293–299 (2004)
  14. Roukas T. Pretreatment of beet molasses to increase pullulan production, Process Biochem, 33(3), 805–810 (1998)
  15. Weifa Z., Bradley S.C., Barbara M.M.D., Robert J.S. Effects of melanin on the accumulation of exopolysaccharides by Aureobasidium pullulans grown on nitrate, Bio res Tech, 99 (3), 7480–7486 (2008)
  16. Yarrow D. Methods for the isolation, maintenance and identification of yeasts. In C.P. Kurtzman & J.W.Fell (Eds.), the yeasts, a taxonomic study, 6, 77-100 (1998)
  17. Yurlova N.A., De Hoog G.S. A new variety of Aureobasidium pullulans characterized by exopolysaccharides structure, nutritional physiology and molecular features, Ant van Leeuwenhoek, 72(3),141–147 (1997)
  18. Zhao S., Chi Z., A New Pullulan-Producing Yeast and Medium Optimization for Its Exopolysaccharide Production, J Ocean Univer Qingdao, 2(3), 53-57 (2003)