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Production of Pullulan by Microbial Fermentation

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Polysaccharides

Abstract

Pullulan is a microbial exopolysaccharide produced by polymorphic fungus Aureobasidium pullulans. Owing to its structure, consisting of α(1→4) and α(1→6) linkages, it has unique properties and hence is suitable for a variety of commercial applications in various field including food, pharmaceuticals, and biomedical. In this chapter, biosynthesis, production through microbial fermentation, and influence of fermentation conditions like pH, temperature, aeration, agitation on microbial growth, pullulan yield, pullulan purity, and molecular weight are discussed. Later, options of using various agricultural wastes as carbon source for pullulan production are considered.

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Abbreviations

ATP:

Adenosine triphosphate

DE:

Dextrose equivalent. It is a measure of reducing sugars present in starch hydrolysate. Carbohydrate molecules like starch are polymers of monosaccharide. The extent of polymerization is indirectly measured by DE. In starch hydrolysate DE represents the amount of reducing sugar as dextrose. Dextrose equivalent of starch is 100 and that of glucose/dextrose is 100. Starch hydrolysates with DE ranging between 1 and 13 are termed as dextrins and those with DE in the range of 13–20 are called maltodextrins. DE for glucose syrup is typically greater than 20, and commercial glucose syrup has a DE of 40–42.

UDPG:

Uridine diphosphate glucose

References

  • Barnett C, Smith A, Scanlon B, Israilides CJ (1999) Pullulan production by Aureobasidium pullulans growing on hydrolysed potato starch waste. Carbohydr Polym 38:203–209

    Article  CAS  Google Scholar 

  • Bernier (1958) The production of polysaccharides by fungi active in the decomposition of wood and forest litter. Can J Microbiol 4:195–204. doi:10.1139/m58-020

    Google Scholar 

  • Campbell BS, Mcdougall BM, Seviour RJ (2003) Why do exopolysaccharide yields from the fungus Aureobasidium pullulans fall during batch culture fermentation? Enzym Microb Technol 33:104–112. doi:10.1016/S0141-0229(03)00089-9

    Article  CAS  Google Scholar 

  • Campbell BS, Siddique AM, Mcdougall BM, Seviour RJ (2004) Which morphological forms of the fungus Aureobasidium pullulans are responsible for pullulan production? FEMS Microbiol Lett 232:225–228. doi:10.1016/S0378-1097(04)00076-X

    Article  CAS  Google Scholar 

  • Cheng K, Demirci A, Catchmark JM (2010a) Effects of plastic composite support and pH profiles on pullulan production in a biofilm reactor. Appl Microbiol Biotechnol 86:853–861. doi:10.1007/s00253-009-2332-x

    Article  CAS  Google Scholar 

  • Cheng K, Demirci A, Catchmark JM (2010b) Enhanced pullulan production in a biofilm reactor by using response surface methodology. J Ind Microbiol Biotechnol 37:587–594. doi:10.1007/s10295-010-0705-x

    Article  CAS  Google Scholar 

  • Cheng K, Demirci A, Catchmark JM, Puri VM (2010c) Modeling of pullulan fermentation by using a color variant strain of Aureobasidium pullulans. J Food Eng 98:353–359. doi:10.1016/j.jfoodeng.2010.01.01

    Google Scholar 

  • Cheng K, Demirci A, Catchmark JM (2011) Pullulan: biosynthesis, production, and applications. Appl Microbiol Biotechnol 92:29–44. doi:10.1007/s00253-011-3477-y

    Article  CAS  Google Scholar 

  • Choudhury AR, Sharma N, Prasad GS (2012) Deoiled jatropha seed cake is a useful nutrient for pullulan production. Microb Cell Factories 11:39. doi:10.1186/1475-2859-11-39

    Article  CAS  Google Scholar 

  • Farris S, Unalan IU, Introzzi L, Fuentes-Alventosa JM, Cozzolino CA (2014) Pullulan-based films and coatings for food packaging: Present applications, emerging opportunities, and future challenges. J App Poly Sci 131: art. no. 40539. doi:10.1002/app.40539

    Google Scholar 

  • Gaidhani HK, Mcneil B, Ni X (2003) Production of pullulan using an oscillatory baffled bioreactor. J Chem Technol Biotechnol 78:260–264. doi:10.1002/jctb.766

    Article  CAS  Google Scholar 

  • Gaidhani HK, McNeil B, Ni X (2005) Fermentation of pullulan using an oscillatory baffled fermenter. Chem Eng Res Des 83:640–645. doi:10.1025/cherd.04355

    Article  CAS  Google Scholar 

  • Gniewosz M, Duszkiewicz-Reinhard W (2008) Comparative studies on pullulan synthesis, melanin synthesis and morphology of white mutant Aureobasidium pullulans B-1 and parent strain A. p. -3. Carbohydr Polym 72:431–438. doi:10.1016/j.carbpol.2007.09.009

    Article  CAS  Google Scholar 

  • Goksungur Y, Uzunogullari P, Dagbagli S (2011) Optimization of pullulan production from hydrolysed potato starch waste by response surface methodology. Carbohydr Polym 83:1330–1337. doi:10.1016/j.carbpol.2010.09.047

    Article  CAS  Google Scholar 

  • Heald PJ, Kristiansen B (1985) Synthesis of polysaccharide by yeast-like forms of Aureobasidium pullulans. Biotechnol Bioeng 27:1516–1519

    Article  CAS  Google Scholar 

  • Israilides CJ, Smith A, Harthill JE, Barnett C, Bambalov G, Scanlon B (1998) Pullulan content of the ethanol precipitate from fermented agro-industrial wastes. Appl Microbiol Biotechnol 49:613–617

    Article  CAS  Google Scholar 

  • Israilides C, Smith A, Scanlon B, Barnett C (1999) Pullulan from agro-industrial wastes. Biotechnol Genet Eng Rev 16:309–315

    Article  CAS  Google Scholar 

  • Ji-Hyun L, Kim J, Kim M, Lim S, Nam S, Lee J, Kim S (2002) Effect of dissolved oxygen concentration and pH on the mass production of high molecular weight pullulan by Aureobasidium pullulans. J Microbiol Biotechnol 12:1–7

    Google Scholar 

  • Kumar D, Saini N, Pandit V, Ali S (2012) An insight to pullulan: a biopolymer in pharmaceutical approaches. Int J Basic Appl Sci 1:202–219

    Google Scholar 

  • Lazaridou A, Roukas T, Biliaderis CG, Vaikousi H (2002) Characterization of pullulan produced from beet molasses by Aureobasidium pullulans in a stirred tank reactor under varying agitation. Enzym Microb Technol 31:122–132

    Article  CAS  Google Scholar 

  • Leathers TD (2003) Biotechnological production and applications of pullulan. Appl Microbiol Biotechnol 62:468–473

    Article  CAS  Google Scholar 

  • Leduy A, Boa JM (1977) Pullulan production from peat hydrolysate. Can J Microbiol 29:143–146

    Article  Google Scholar 

  • Li B, Zhang N, Peng Q, Yin T, Guan F, Wang G, Li Y (2009) Production of pigment-free pullulan by swollen cell in Aureobasidium pullulans NG which cell differentiation was affected by pH and nutrition. Appl Microbiol Biotechnol 84:293–300. doi:10.1007/s00253-009-1955-2

    Article  CAS  Google Scholar 

  • Ma Z, Fu W, Liu G, Wang Z-P, Chi Z-M (2014) High-level pullulan production by Aureobasidium pullulans var. melanogenium P16 isolated from mangrove system. Appl Microbiol Biotechnol 98:4865–4873. doi:10.1007/s00253-014-5554-5

    Article  CAS  Google Scholar 

  • Mehta A, Prasad GS, Choudhury AR (2014) Cost effective production of pullulan from agri-industrial residues using response surface methodology. Int J Biol Macromol 64:252–256. doi:10.1016/j.ijbiomac.2013.12.011

    Article  CAS  Google Scholar 

  • Moubasher H, Wahsh SS, Abo NE-K (2013) Isolation of Aureobasidium pullulans and the effect of different conditions for pullulanase and pullulan production. Microbiology 82:155–161. doi:10.1134/S0026261713020197

    Article  CAS  Google Scholar 

  • Oguzhan P, Yangilar F (2013) Pullulan: production and usage in food industry. Afr J Food Sci Technol 4:57–63

    Google Scholar 

  • Ray RC, Moorthy SN (2007) Exopolysaccharide (pullulan) production from cassava starch residue by Aureobasidium pullulans strain MTTC 1991. J Sci Ind Res 66:252–255

    CAS  Google Scholar 

  • Roukas T (1998) Pretreatment of beet molasses to increase pullulan production. Process Biochem 33:805–810

    Article  CAS  Google Scholar 

  • Roukas T (1999) Pullulan production from deproteinized whey by Aureobasidium pullulans. J Ind Microbiol Biotechnol 22:617–621

    Article  CAS  Google Scholar 

  • Roukas T, Biliaderis CG (1995) Evaluation of carob pod as a substrate for pullulan production by Aureobasidium pullulans. Appl Biochem Biotechnol 55:27–44

    Article  CAS  Google Scholar 

  • Sena RF, Costelli MC, Gibson LH, Coughlin RW (2006) Enhanced production of pullulan by two strains of A. pullulans with different concentrations of soybean oil in sucrose solution IN batch fermentations. Braz J Chem Eng 23:507–515

    Article  CAS  Google Scholar 

  • Sevious RJ, Stasinopoulos SJ, Auer DPF, Gibbs PA (1992) Production of pullulan and other exopolysaccharides by filamentous fungi. Crit Rev Biotechnol 12:279–298

    Article  Google Scholar 

  • Sheng L, Zhu G, Tong Q (2014) Effect of uracil on pullulan production by Aureobasidium pullulans. Carbohydr Polym 101:435–437. doi:10.1016/j.carbpol.2013.09.063

    Article  CAS  Google Scholar 

  • Shingel KI (2004) Current knowledge on biosynthesis, biological activity, and chemical modification of the exopolysaccharide, pullulan. Carbohydr Res 339:447–460. doi:10.1016/j.carres.2003.10.034

    Article  CAS  Google Scholar 

  • Simon L, Bouchonneau M (1993) Relation between pullulan production, morphological state and growth conditions in Aureobasidium pullulans: new observations. J Gen Microbiol 139:979–985

    Article  CAS  Google Scholar 

  • Singh RS, Saini GK (2008) Pullulan-hyperproducing color variant strain of Aureobasidium pullulans FB-1 newly isolated from phylloplane of Ficus sp. Bioresour Technol 99:3896–3899. doi:10.1016/j.biortech.2007.08.003

    Article  CAS  Google Scholar 

  • Singh RS, Saini GK, Kennedy JF (2008) Pullulan: microbial sources, production and applications. Carbohydr Polym 73:515–531. doi:10.1016/j.carbpol.2008.01.003

    Article  CAS  Google Scholar 

  • Singh RS, Saini GK, Kennedy JF (2009) Downstream processing and characterization of pullulan from a novel colour variant strain of Aureobasidium pullulans FB-1. Carbohydr Polym 78:89–94. doi:10.1016/j.carbpol.2009.03.040

    Article  CAS  Google Scholar 

  • Singh R, Gaur R, Tiwari S, Gaur MK (2012) Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process. Braz J Microbiol 43:1042–1050

    Article  CAS  Google Scholar 

  • Sugumaran KR, Gowthami E, Swathi B, Elakkiya S, Srivastava SN, Ravikumar R, Gowdhaman D, Ponnusami V (2013a) Production of pullulan by Aureobasidium pullulans from Asian palm kernel: a novel substrate. Carbohydr Polym 92:697–703. doi:10.1016/j.carbpol.2012.09.062

    Article  CAS  Google Scholar 

  • Sugumaran KR, Sindhu RV, Sukanya S, Aiswarya N, Ponnusami V (2013b) Statistical studies on high molecular weight pullulan production in solid state fermentation using jack fruit seed. Carbohydr Polym 98:854–860. doi:10.1016/j.carbpol.2013.06.071

    Article  CAS  Google Scholar 

  • Survase SA, Saudagar PS, Bajaj IB, Singhal RS (2007) Scleroglucan: fermentative production, downstream processing and applications. Food Technol Biotechnol 45:107–118

    CAS  Google Scholar 

  • Thirumavalavan K, Manikkadan TR, Dhanasekar R (2009) Pullulan production from coconut by-products by Aureobasidium pullulans. Afr J Biotechnol 8:254–258

    CAS  Google Scholar 

  • Wang D, Yu X, Gongyuan W (2013) Pullulan production and physiological characteristics of Aureobasidium pullulans under acid stress. Biotechnol Prod Process Eng 97:8069–8077. doi:10.1007/s00253-013-5094-4

    CAS  Google Scholar 

  • Wecker A, Onken U (1991) Influence of dissolved oxygen concentration and shear rate on the production of pullulan by Aureobasidium pullulans. Biotechnol Lett 13:155–160

    Article  CAS  Google Scholar 

  • Wu S, Jin Z, Tong Q, Chen H (2009) Sweet potato: a novel substrate for pullulan production by Aureobasidium pullulans. Carbohydr Polym 76:645–649. doi:10.1016/j.carbpol.2008.11.034

    Article  CAS  Google Scholar 

  • Wu S, Chen H, Jin Z (2010) Effect of two-stage temperature on pullulan production by Aureobasidium pullulans. World J Microbiol Biotechnol 1234:737–741. doi:10.1007/s11274-009-0231-z

    Article  Google Scholar 

  • Xia Z, Wu S, Pan S (2011) Effect of two-stage controlled pH and temperature on pullulan production by Aureobasidium pullulans. Carbohydr Polym 86:1814–1816. doi:10.1016/j.carbpol.2011.06.087

    Article  CAS  Google Scholar 

  • Youssef F, Biliaderis CG, Roukas T (1998) Enhancement of pullulan production by Aureobasidium pullulans in batch culture using olive oil and sucrose as carbon sources. Appl Biochem Biotechnol 74:13–30

    Article  CAS  Google Scholar 

  • Youssef F, Roukas T, Biliaderis CG (1999) Pullulan production by a nonpigmented strain of Aureobasidium pullulans using batch and fed-batch culture. Process Biochem 34:355–366

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. School of Chemical and Biotechnology, SASTRA University, Thanjavur, TN, 613401, India

    V. Ponnusami & V. Gunasekar

Authors
  1. V. Ponnusami
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  2. V. Gunasekar
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Correspondence to V. Ponnusami .

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Editors and Affiliations

  1. Botany Department, M.L.Sukhadia University, Badgaon, India

    Kishan Gopal Ramawat

  2. Institut des Sciences de la Vigne et du Vin, Université de Bordeaux, Villenave d'Ornon, France

    Jean-Michel Mérillon

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Ponnusami, V., Gunasekar, V. (2014). Production of Pullulan by Microbial Fermentation. In: Ramawat, K., Mérillon, JM. (eds) Polysaccharides. Springer, Cham. https://doi.org/10.1007/978-3-319-03751-6_58-1

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  • DOI: https://doi.org/10.1007/978-3-319-03751-6_58-1

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  • Publisher Name: Springer, Cham

  • Online ISBN: 978-3-319-03751-6

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