Abstract
Objective
Fructooligosaccharides (FOS) are prebiotic substances that have been extensively incorporated in different products of food industry mostly for their bifidogenic properties and economic value. The main commercial FOS production comes from the biotransformation of sucrose and intracellular and extracellular microbial enzymes–fructosyltransferases (FTase). Aspergillus oryzae IPT-301 produces FTase. In order to increase its production, this study focuses on evaluating the effects of different agitation speed and aeration rates which affect yields in a stirred tank bioreactor.
Results
Agitation had more influence on cell growth than aeration. The maximum intracellular FTase activity and the volumetric productivity of total intracellular FTase were obtained at 800 rpm and 0.75 vvm, and reached values of 2100 U g−1 and 667 U dm−3 h−1, respectively. The agitation speed had a strong influence on the activity of extracellular FTase produced which reached the maximum amount of 53 U cm−3. The higher value of total activity obtained was 22,831 U dm−3 at 0.75 vvm and 800 rpm.
Conclusion
Aeration rates and agitation speed showed strong influence upon the growth and production of fructosyltransferase from Aspergillus oryzae IPT-301 in media containing sucrose as carbon source. The control of aeration rate and agitation speed can be a valuable fermentation strategy to improve enzyme production.
Similar content being viewed by others
References
Albaek MO, Gernaey KV, Hansen MS, Stocks SM (2011) Modeling enzyme production with Aspergillus oryzae in pilot scale vessels with different agitation, aeration, and agitator types. Biotechnol Bioeng 108:1828–1840. https://doi.org/10.1002/bit.23121
Antosŏvá M, Polakovič M, Slovinská M, Madlová A, Illeová V, Baleš V (2002) Effect of sucrose concentration and cultivation time on batch production of fructosyltransferase by Aureobasidium pullulans CCY 27-1-1194. Chem Pap 56(6):394–399
Antosová M, Polakovic M (2001) Fructosyltransferases: the enzymes catalyzing production of fructooligosaccharides. Chem Pap 55(6):350–358
Antosová M, Illeová V, Vandáková M, Druzkovská A, Polakovic M (2008) Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans. J Biotechnol 135:58–63. https://doi.org/10.1016/j.jbiotec.2008.02.016
Balasubramaniem AK, Nagarajan KV, Paramasamy G (2001) Optimization of media for β-fructofuranosidase production by Aspergillus niger in submerged and solid-state fermentation. Process Biochem 36(12):1241–1247. https://doi.org/10.1016/S0032-9592(01)00166-2
Bartholomew WH, Karow EO, Sfat MR (1950) Oxygen transfer and agitation in submerged fermentations: mass transfer of oxygen in submerged fermentation of Streptomycin griseus. Ind Eng Chem 42(9):1801–1809. https://doi.org/10.1021/ie50489a032
Chen WC, Liu CH (1996) Production of β-fructofuranosidase by Aspergillus japonicus. Enzyme Microb Technol 18:153–160. https://doi.org/10.1016/0141-0229(95)00099-2
Cruz R, Cruz VD, Belini MZ, Belote J, Vieira CR (1998) Production of fructooligosaccharides by the mycelia of Aspergillus japonicus immobilized in calcium alginate. Bioresour Technol 65:139–143. https://doi.org/10.1016/S0960-8524(98)00005-4
Cuervo R, Guilarte B, Juárez A, Martínez J (2004) Production of fructooligosaccharides by β-fructofuranosidase from Aspergillus sp. 27H. J Chem Technol Biotechnol 79:268–272. https://doi.org/10.1002/jctb.967
Cuervo-Fernández R, Ottoni CA, Silva ES, Matsubara RMS, Carter JM, Magossi LR, Wada MAA, Rodrigues MFA, Maresma BG, Maiorano AE (2007) Screening of β-fructofuranosidase producing microorganisms and effect of pH and temperature on enzymatic rate. Appl Microbiol Biotechnol 75:87–93. https://doi.org/10.1007/s00253-006-0803-x
Cunha JS, Ottoni CA, Morales SAV, Silva ES, Maiorano AE, Perna RF (2019) Synthesis and characterization of fructosyltransferase from Aspergillus oryzae IPT-301 for high fructooligosaccharides production. Braz J Chem Eng 36(2):657–668. https://doi.org/10.1590/0104-6632.20190362s20180572
Dhake AB, Patil MB (2007) Effect of substrate feeding on production of fructosyltransferase by Penicillium purpurogenum. Braz J Microbiol 38:194–199. https://doi.org/10.1590/S1517-83822007000200002
Dominguez A, Nobre C, Rodrigues LR, Peres AM, Torres D, Rocha I (2012) New improved method for fructooligosaccharides production by Aureobasidium pullulans. Carbohydr Polym 89(4):1174–1179. https://doi.org/10.1016/j.carbpol.2012.03.091
Dorta C, Cruz R, Neto PO, Moura DJC (2006) Sugarcane molasses and yeast powder used in the fructooligosaccharides production by Aspergillus japonicus-FCL 119T and Aspergillus niger ATCC 20611. J Ind Microbiol Biot 33:1003–1009. https://doi.org/10.1007/s10295-006-0152-x
Fenice M, Barghini P, Selbmann L, Federici F (2012) Combined effects of agitation and aeration on the chitinolytic enzymes production by the Antarctic fungus Lecanicillium muscarium CCFEE 5003. Microb Cell Fact 11:1–10. https://doi.org/10.1186/1475-2859-11-12
Flores-Maltos DA, Mussatto SI, Contreras-Esquivel JC, Rodríguez-Herrera R, Teixeira JA, Aguilar CN (2016) Biotechnological production and application of fructooligosaccharides. Crit Rev Biotechnol 36(2):259–267. https://doi.org/10.3109/07388551.2014.953443
Ganaie MA, Gupta US, Kango N (2013) Screening of biocatalysts for transformation of sucrose to fructooligosaccharides. J Mol Catal B 97:12–17. https://doi.org/10.1016/j.molcatb.2013.07.008
Ganaie MA, Lateef A, Gupta US (2014) Enzymatic trends of fructooligosaccharides production by microorganisms. Appl Biochem Biotechnol 172:2143–2159. https://doi.org/10.1007/s12010-013-0661-9
Garcia-Ochoa F, Gomez E (2009) Bioreactor scale-up and oxygen transfer rate in microbial processes: an overview. Biotecnol Adv 27:153–176. https://doi.org/10.1016/j.biotechadv.2008.10.006
Grimm LH, Kelly S, Krull R, Hempel DC (2005) Morphology and productivity of filamentous fungi. Appl Microbiol Biotechnol 69:375–384. https://doi.org/10.1007/s00253-005-0213-5
Gupta AK, Bhatia IS (1982) Glucofructosan biosynthesis in Fusarium oxysporum regulation and substrate specificity of fructosyltransferase and invertase. Phytochemistry 21:1249–1253
Hayashi S, Matsuzaki K, Takasaki Y, Ueno H, Imada K (1992) Production of β-fructofuranosidase by Aspergillus japonicus. World J Microb Biotechnol 8(2):155–159. https://doi.org/10.1007/BF01195837
Hidaka H, Hirayama M, Sumi N (1988) Fructooligosaccharide-producing enzyme from Aspergillus niger ATCC 20611. Agric Biol Chem 52(5):1181–1187. https://doi.org/10.1080/00021369.1988.10868810
Jitonnom J, Ketudat-Cairns JR, Hannongbua S (2018) QM/MM modeling of the hydrolysis and transfructosylation reactions of fructosyltransferase from Aspergillus japonicas, an enzyme that produces prebiotic fructooligosaccharide. J Mol Graph Model 79:175–184. https://doi.org/10.1016/j.jmgm.2017.11.010
L’Hocine L, Jiang ZWB, Xu S (2000) Purification and partial characterization of fructosyltransferase and invertase from Aspergillus niger AS0023. J Biotechnol 81:73–84. https://doi.org/10.1016/S0168-1656(00)00277-7
Lateef A, Oloke JK, Gueguim-Kana EB, Raimi OR (2012) Production of fructosyltransferase by a local isolate of Aspergillus niger in both submerged and solid substrate. Acta Aliment Hung 41:100–117. https://doi.org/10.1556/AAlim.41.2012.1.12
Lim JS, Park MC, Lee JH, Park SW, Kim SW (2005) Optimization of culture medium and conditions for Neo-fructooligosaccharides production by Penicillium citrinum. Eur Food Res Technol 221:639–644. https://doi.org/10.1007/s00217-005-0070-6
Lim JS, Lee JH, Kim JM, Park SW, Kim SW (2006) Effects of morphology and rheology on neo-fructosyltransferase production by Penicillium citrinum. Biotechnol Bioprocess Eng 11:100–104. https://doi.org/10.1007/BF02931891
Liu BL, Kao PM, Tzeng YM, Feng KC (2003) Production of chitinase from Verticillium lecanii F091 using submerged fermentation. Enzyme Microb Technol 33:410–415. https://doi.org/10.1016/S0141-0229(03)00138-8
Maiorano AE, Piccoli RM, Silva ES, Rodrigues MFA (2008) Microbial production of fructosyltransferases for synthesis of pre-biotics. Biotechnol Lett 30(11):1867–1877. https://doi.org/10.1007/s10529-008-9793-3
Muñiz-Márquez DB, Contreras JC, Rodríguez R, Mussatto SI, Teixeira JA, Aguilar CN (2016) Enhancement of fructosyltransferase and fructooligosaccharides production by A. oryzae DIA-MF in solid-state fermentation using aguamiel as culture medium. Bioresour Technol 213:276–282. https://doi.org/10.1016/j.biortech.2016.03.022
Mussatto SI, Ballesteros LF, Silvia Martins S, Maltos DAF, Aguilar CN, Teixeira JA (2012) Maximization of fructooligosaccharides and β-fructofuranosidase production by Aspergillus japonicus under solid-state fermentation conditions. Food Bioprocess Techol 6:2128–2134. https://doi.org/10.1007/s11947-012-0873-y
Nguyen QD, Szabó JMR, Bhat MK, Hoschke AP (2005) Purification and some properties of β-fructofuranosidase from Aspergillus niger IMI303386. Process Biochem 40:2461–2466. https://doi.org/10.1016/j.procbio.2004.09.012
Ottoni CA, Cuervo-Fernández R, Piccoli RM, Moreira R, Guilarte-Maresma B, Silva ES, Rodrigues MFA, Maiorano AE (2012) Media optimization for β-fructofuranosidase for production by Aspergillus oryzae. Braz J Chem Eng 29(01):49–59. https://doi.org/10.1590/S0104-66322012000100006
Palma MB, Milagres AMF, Prata AMR, Mancilha IM (1996) Influence of aeration and agitation rate on the xylanase activity from Penicillium janthinellum. Process Biochem 31:141–145. https://doi.org/10.1016/0032-9592(95)00042-9
Park JP, Oh TK, Yun JW (2001) Purification and characterization of a novel transfructosylating enzyme from Bacillus macerans EG-6. Process Biochem 37:471–476. https://doi.org/10.1016/S0032-9592(01)00237-0
Patel V, Saunders G, Bucke C (1997) N-Deglycosylation of fructosyltransferase and invertase from Fusarium oxysporum decreases stability but has little effect on kinetics and synthetic specificity. Biotechnol Lett 19:75–77. https://doi.org/10.1023/A:1018375222571
Perna RF, Cunha JS, Gonçalves MCP, Basso RC, Silva ES, Maiorano AE (2018) Microbial fructosyltransferase: production by submerged fermentation and evaluation of pH and temperature effects on transfructosylation and hydrolytic enzymatic activities. Int J Eng Res Sci 4(3):43–50. https://doi.org/10.5281/zenodo.1213538
Sangeetha PT, Ramesh MN, Prapulla SG (2004a) Production of fructooligosaccharides by fructosyltransferase from Aspergillus oryzae CFR 202 and Aureobasidium pullulans CFR 77. Process Biochem 39:753–758. https://doi.org/10.1016/S0032-9592(03)00186-9
Sangeetha PT, Ramesh MN, Prapulla SG (2004b) Production of fructosyltransferase by Aspergillus oryzae CFR 202 in solid-state fermentation using agricultural by-products. Appl Microbiol Biotechnol 65:530–537. https://doi.org/10.1007/s00253-004-1618-2
Sangeetha PT, Ramesh MN, Prapulla SG (2005) Fructooligosaccharide production using fructosyltransferase obtained from recycling culture of Aspergillus oryzae CFR 202. Process Biochem 40(34):1085–1088. https://doi.org/10.1016/j.procbio.2004.03.009
Schell DJ, Farmer J, Hamilton J, Lyons B, Mcmillan JD, Sáez JC, Tholudur A (2001) Influence of operating conditions and vessel size on oxygen transfer during cellulase production. Appl Biochem Biotechnol 91–93:627–642. https://doi.org/10.1385/ABAB:91-93:1-9:627
Šedová M, Illeová V, Antošová M, Annus J, Polakovič M (2014) Production of fructosyltransferase in mechanically stirred and air-lift bioreactors. Chem Pap 68(12):1639–1648. https://doi.org/10.2478/s11696-014-0563-5
Serrano-Carreon L, Corona RM, Sfinchez A, Galindo E (1998) Prediction of xanthan fermentation development by a model linking kinetics, power drawn and mixing. Process Biochem 33(2):133–146. https://doi.org/10.1016/S0032-9592(97)00039-3
Silva-Santisteban BOY, Maugeri F (2005) Agitation, aeration and shear stress as key factors in inulinase production by Kluyveromyces marxianus. Enzyme Microb Technol 36:717–724. https://doi.org/10.1016/j.enzmictec.2004.12.008
Techapun C, Poosaran N, Masanori W, Sasaki K (2003) Optimization of aeration and agitation rates to improve cellulose-free xylanase production by thermotolerant Streptomyces sp. Ab 106 and repeated fed-batch cultivation using agricultural waste. J Biosci Bioeng 95(3):298–301. https://doi.org/10.1016/S1389-1723(03)80033-6
Wang XD, Rakshit SK (1999) Improved extracellular transferase enzyme production by Aspergillus foetidus for synthesis of isooligosaccharides. Bioprocess Eng 20:429–434. https://doi.org/10.1007/PL00009053
Wang LM, Zhou HM (2006) Isolation and identification of a novel Aspergillus japonicus JN19 producing β-fructofuranosidase and characterization of the enzyme. J Food Biochem 30:641–658. https://doi.org/10.1111/j.1745-4514.2006.00085.x
Yun JW, Kim DH, Kim BW, Song SK (1997) Comparison of sugar compositions between inulo- and fructo-oligosaccharides produced by different enzyme forms. Biotechnol Lett 19:553–556. https://doi.org/10.1023/A:101839350
Zhang J, Liu C, Xie Y, Li N, Ning Z, Du N, Huang X, Zhong Y (2017) Enhancing fructooligosaccharides production by genetic improvement of the industrial fungus Aspergillus niger ATCC 2061. J Biotechnol 249:25–33. https://doi.org/10.1016/j.jbiotec.2017.03.021
Acknowledgements
The authors gratefully acknowledge the financial support of the São Paulo Research Foundation (FAPESP).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Maiorano, A.E., da Silva, E.S., Perna, R.F. et al. Effect of agitation speed and aeration rate on fructosyltransferase production of Aspergillus oryzae IPT-301 in stirred tank bioreactor. Biotechnol Lett 42, 2619–2629 (2020). https://doi.org/10.1007/s10529-020-03006-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-020-03006-9