Abstract
Product formation of mycelial organisms, like Aspergillus niger, is intimately connected with their morphology. Pellet morphology is often requested for product formation. Therefore, it is important to reveal the influence of the hydrodynamic conditions on the morphological development. In the present study, pellet morphology and glucoamylase formation were studied under different agitation intensities of A. niger AB1.13. For pellet formation inside the bioreactor, without the use of precultures, it is necessary to work at low energy dissipation rates. Biomass growth and glucoamylase activity were correlated with energy dissipation. Furthermore, product yield was analysed in dependence of pellet size and concentration. The present work shows that simple equations based on Monod-kinetics can describe growth and product formation, in general, also in mycelian organisms. All measured morphological data, like pellet concentration, as well as glucoamylase formation, strongly depend on the hydrodynamic conditions.
Similar content being viewed by others
Abbreviations
- α :
-
Growth-associated product formation coefficient (μKat g−1)
- β :
-
Non-growth-associated product formation coefficient (μKat g−1h−1)
- ε :
-
Energy dissipation (W kg−1)
- μ :
-
Specific growth rate (h−1)
- μ max :
-
Maximum specific growth rate (h−1)
- ρ P :
-
Density of wet pellet (kg m−3)
- EA GAM :
-
Glucoamylase enzyme activity in the bulk phase (μKat l−1)
- C Glu :
-
Glucose concentration (g l−1)
- \( C_{{{\text{O}}_{2} }} \) :
-
Oxygen concentration (bulk) (g l−1)
- \( C^{ * }_{{{\text{O}}_{2} }} \) :
-
Saturation concentration of oxygen (bulk) (g l−1)
- C P :
-
Pellet concentration (l−1)
- d P :
-
Pellet diameter (mm)
- K Glu :
-
Saturation coefficient glucose (g l−1)
- \( K_{{{\text{O}}_{2} }} \) :
-
Saturation coefficient oxygen (g l−1)
- k L a :
-
Volumetric gas liquid mass transfer coefficient (h−1)
- R GAM :
-
Glucoamylase production rate (μKat l−1h−1)
- R Glu :
-
Glucose consumption rate (g l−1h−1)
- \( R_{{{\text{O}}_{2} }} \) :
-
Oxygen consumption rate (g l−1h−1)
- R X :
-
Biomass growth rate (g l−1h−1)
- X :
-
Biomass concentration (g l−1)
- Y OX :
-
Yield coefficient of biomass on oxygen (g g−1)
- Y SX :
-
Yield coefficient of biomass on glucose (g g−1)
References
Amanullah A, Jüsten P, Davies A, Paul GC, Nienow AW, Thomas CR (2000) Agitation induced mycelial fragmentation of Aspergillus oryzae and Penicillium chrysogenum. Biochem Eng J 5:109–114
Amanullah A, Leonildi E, Nienow AW, Thomas CR (2001) Dynamics of mycelia aggregation in cultures of Aspergillus oryzae. Bioproc Biosyst Eng 24:101–107
Bhargava S, Wenger K, Marten M (2003) Pulsed addition of limiting-carbon during Aspergillus oryzae fermentation leads to improved productivity of a recombinant enzyme. Biotechnol Bioeng 85(1):111–117
Braun S, Vecht-Lifshitz SE (1991) Mycelial morphology and metabolite production. Trends Biotechnol 9:63–68
Cox PW, Thomas CR (1992) Classification and measurement of fungal pellets by automated image analysis. Biotechnol Bioeng 39:945–952
Cui YQ, van der Lans RGJM, Luyben KCAM (1997) Effect of agitation intensities on fungal morphology of submerged fermentation. Biotechnol Bioeng 55(5):715–726
Cui YQ, van der Lans RGJM, Giuseppin MLF, Luyben KCAM (1998) Influence of fermentation conditions and scale on the submerged fermentation of Aspergillus awamori. Enz Micro Tech 23:157–167
Cui YQ, van der Lans RGJM, Luyben KCAM (1998) Effects of dissolved oxygen tension and mechanical forces on fungal morphology in submerged fermentation. Biotechnol Bioeng 57(4):409–419
Cui YQ, Okkerse WJ, van der Lans RGJM, Luyben KCAM (1998) Modeling and measurements of fungal growth and morphology in submerged fermentations. Biotechnol Bioeng 60(2):216–229
Davies JL, Baganz F, Ison AP, Lye GJ (2000) Studies on the interaction of fermentation and microfiltration operations: erythromycin recovery from Saccharopolyspora erythraea fermentation broths. Bioproc Eng 69(4):429–439
Dynesen J, Nielsen J (2003) Surface hydrophobicity of Aspergillus nidulans conidiospores and its role in pellet formation. Biotechnol Prog 19(3):1049–1052
El-Enshasy HA (1998) Optimization of glucose oxidase production and excretion by recombinant Aspergillus niger. Technische Universität Carolo-Wilhelmina, Braunschweig
El-Enshasy HA, Hellmuth K, Rinas U (1999) Fungal morphology in submerged cultures and its relation to glucose oxidase excretion by recombinant Aspergillus niger. Appl Biochem Biotechnol 81(1):1–11
Fujita M, Iwahori KST, Yamakawa K (1994) Analysis of pellet formation of Aspergillus niger based on shear-stress. J Ferment Bioeng 78(5):368–373
Ganzlin M (2000) Untersuchungen der induzierten Proteinproduktion unter Kontrolle des Glucoamylasepromotors in Aspergillus niger. Technische Universität Carolo-Wilhelmina, Braunschweig
Grimm LH, Kelly S, Hengstler J, Göbel A, Krull R, Hempel DC (2003) Kinetic studies on the aggregation of Aspergillus niger conidia. Biotechnol Bioeng 87(2):213–218
Hellendoorn L, Mulder H, van den Heuvel JC, Ottengraf SPP (1998) Intrinsic kinetic parameters of the pellet forming fungus Aspergillus awamori. Biotechnol Bioeng 58(5):478–485
Johansen CL, Coolen L, Hunik JH (1998) Influence of morphology on product formation in Aspergillus awamori during submerged fermentations. Biotechnol Prog 14:233–240
Koutinas AA, Wang R, Webb C (2003) Estimation of fungal growth in complex, heterogeneous culture. Biochem Eng J 14:93–100
Li ZJ, Shukla V, Fordyce AP, Pedersen AG, Wenger KS, Marten MR (2000) Fungal morphology and fragmentation behavior in a fed-batch Aspergillus oryzae fermentation at the production scale. Biotechnol Bioeng 70(3):300–312
Liu F, Li W, Ridgway D, Gu T, Moo-Young M (1998) Inhibition of extracellular protease secretion by Aspergillus niger using cell immobilization. Biotechnol Lett 20(6):539–542
Liu J-Z, Weng L-P, Zhang Q-L, Xu H, Ji L-N (2003) A mathematical model for gluconic acid fermentation by Aspergillus niger. Biochem Eng J 14:137–141
Mahnke EU (2001) Fluiddynamisch induzierte Partikelbeanspruchung in pneumatisch gerührten Mehrphasenreaktoren. In: Hempel DC (ed) ibvt-Schriftenreihe, vol 12. FIT-Verlag: Paderborn as well as PhD thesis, Technische Universität Carolo-Wilhelmina zu Braunschweig
Mattern IE, van Noort JM, van den Berg P, Archer DB, Roberts IN, van den Hondel CAMJJ (1992) Isolation and characterization of mutants of Aspergillus niger deficient in extracellular proteases. Mol Gen Genomics 234:332–336
Metz B, Kossen NWF (1977) The growth of molds in the form of pellets—a literature review. Biotechnol Bioeng 14:781–799
Mitard A, Riba JP (1988) Morphology and growth of Aspergillus niger ATCC 26036 cultivated at several shear rates. Biotechnol Bioeng 32:835–840
Nielsen J, Johansen CL, Jacobsen M, Krabben P, Villadsen J (1995) Pellet formation and fragmentation in submerged cultures of Penicillium chrysogenum and its relation to penicillin production. Biotechnol Prog 11(1):93–98
Papagianni M, Mattey M, Kristiansen B (1998) Citric acid production and morphology of Aspergillus niger as functions of the mixing intensity in a stirred tank and a tubular loop bioreactor. Biochem Eng J 2:197–205
Paul GC, Priede MA, Thomas CR (1999) Relationship between morphology and citric acid production in submerged Aspergillus niger fermentations. Biochem Eng J 3:121–129
Punt PJ, van Biezen N, Conesa A, Albers AJM, van den Hondel CAMJJ (2002) Filamentous fungi as cell factories for heterologous protein production. Trends Biotechnol 20(5):200–206
Sauer T (1986) Einfluß inerter Partikel auf die Hydrodynamik und den volumetrischen Stoffübergangskoeffizienten in einer Suspensions-Blasensäule, Uni-GH Paderborn
Withers JM, Swift RJ, Wiebe MG, Robson GD, Punt PJ, van den Hondel CAMJJ, Trinci APJ (1998) Optimization and stability of glucoamylase production by recombinant strains of Aspergillus niger in chemostat culture. Biotechnol Bioeng 59(4):407–418
Wongwicharn A, Harvey LM, McNeil B (1999) Secretion of heterologous and native proteins, growth and morphology in batch cultures of Aspergillus niger B1-D at varying agitation rates. J Chem Technol Biotechnol 74:821–828
Zetelaki K, Vas K (1968) The role of aeration and agitation in the production of glucose oxidase in submerged culture. Biotechnol Bioeng 10:45–59
Acknowledgements
The authors acknowledge financial support provided by the German Research Foundation. This project is part of the SFB 578 “Development of biotechnological processes by integrating genetic and engineering methods—from gene to product”.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kelly, S., Grimm, L.H., Hengstler, J. et al. Agitation effects on submerged growth and product formation of Aspergillus niger. Bioprocess Biosyst Eng 26, 315–323 (2004). https://doi.org/10.1007/s00449-004-0368-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-004-0368-y