Switching industrial production processes from complex to defined media: method development and case study using the example of Penicillium chrysogenum
Filamentous fungi are versatile cell factories and widely used for the production of antibiotics, organic acids, enzymes and other industrially relevant compounds at large scale. As a fact, industrial production processes employing filamentous fungi are commonly based on complex raw materials. However, considerable lot-to-lot variability of complex media ingredients not only demands for exhaustive incoming components inspection and quality control, but unavoidably affects process stability and performance. Thus, switching bioprocesses from complex to defined media is highly desirable.
This study presents a strategy for strain characterization of filamentous fungi on partly complex media using redundant mass balancing techniques. Applying the suggested method, interdependencies between specific biomass and side-product formation rates, production of fructooligosaccharides, specific complex media component uptake rates and fungal strains were revealed. A 2-fold increase of the overall penicillin space time yield and a 3-fold increase in the maximum specific penicillin formation rate were reached in defined media compared to complex media.
The newly developed methodology enabled fast characterization of two different industrial Penicillium chrysogenum candidate strains on complex media based on specific complex media component uptake kinetics and identification of the most promising strain for switching the process from complex to defined conditions. Characterization at different complex/defined media ratios using only a limited number of analytical methods allowed maximizing the overall industrial objectives of increasing both, method throughput and the generation of scientific process understanding.
- Meyer V: Genetic engineering of filamentous fungi - Progress, obstacles and future trends. Biotechnol Adv 2008,26(2):177–185. CrossRef
- Papagianni M: Fungal morphology and metabolite production in submerged mycelial processes. Biotechnol Adv 2004,22(3):189–259. CrossRef
- Junker BH, Hesse M, Burgess B, Masurekar P, Connors N, Seeley A: Early phase process scale-up challenges for fungal and filamentous bacterial cultures. Appl Biochem Biotechnol 2004,119(3):241–277. CrossRef
- Nielsen J: Physiological Engineering Aspects of Penicillium chrysogenum. World Scientific, Singapore; 1995.
- Jose GE, Folque F, Menezes JC, Werz S, Strauss U, Hakemeyer C: Predicting Mab product yields from cultivation media components, using near-infrared and 2D-fluorescence spectroscopies. Biotechnol Prog 2011,27(5):1339–1346. CrossRef
- ICH: Q8, Pharmaceutical Development. http://www.ich.org
- Zhang G, Block DE: Using highly efficient nonlinear experimental design methods for optimization of Lactococcus lactis fermentation in chemically defined media. Biotechnol Prog 2009,25(6):1587–1597.
- Zhang G, Mills DA, Block DE: Development of chemically defined media supporting high-cell-density growth of lactococci, enterococci, and streptococci. Appl Environ Microbiol 2009,75(4):1080–1087. CrossRef
- Zhang J, Zhou J, Liu J, Chen K, Liu L, Chen J: Development of chemically defined media supporting high cell density growth of Ketogulonicigenium vulgare and Bacillus megaterium. Bioresour Technol 2009,102(7):4807–4814. CrossRef
- Song H, Kim TY, Choi B-K, Choi SJ, Nielsen LK, Chang HN, Lee SY: Development of chemically defined medium for Mannheimia succiniciproducens based on its genome sequence. Appl Microbiol Biotechnol 2008,79(2):263–272. CrossRef
- Hull SR, Yang BY, Venzke D, Kulhavy K, Montgomery R: Composition of Corn Steep Water during Steeping. J Agric Food Chem 1996,44(7):1857–1863. CrossRef
- De Kok HE, Roels JA: Method for the statistical treatment of elemental and energy balances with application to steady-state continuous-culture growth of Saccharomyces cerevisiae CBS 426 in the respiratory region. Biotechnol Bioeng 1980,22(5):1097–1104. CrossRef
- Wang NS, Stephanopoulos G: Application of macroscopic balances to the identification of gross measurement errors. Biotechnol Bioeng 1983, 25:2177–2208. CrossRef
- Herwig C, Marison I, Von Stockar U: On-line stoichiometry and identification of metabolic state under dynamic process conditions. Biotechnol Bioeng 2001,75(3):345–354. CrossRef
- Jobe AM, Herwig C, Surzyn M, Walker B, Marison I, von Stockar U: Generally applicable fed-batch culture concept based on the detection of metabolic state by on-line balancing. Biotechnol Bioeng 2003,82(6):627–639. CrossRef
- Wechselberger P, Seifert A, Herwig C: PAT method to gather bioprocess parameters in real-time using simple input variables and first principle relationships. Chem Eng Sci 2010,65(21):5734–5746. CrossRef
- Roels JA: Application of macroscopic principles to microbial metabolism. Biotechnol Bioeng 1980,22(12):2457–2514. CrossRef
- van der Heijden RTJM, Romein B, Heijnen JJ, Hellinga C, Luyben KCAM: Linear constraint relations in biochemical reaction systems: I. Classification of the calculability and the balanceability of conversion rates. Biotechnol Bioeng 1994, 43:3–20. CrossRef
- van der Heijden RTJM, Romein B, Heijnen JJ, Hellinga C, Luyben KCAM: Linear constraint relations in biochemical reaction systems: II. Diagnosis and estimation of gross errors. Biotechnol Bioeng 1994, 43:11–20. CrossRef
- van der Heijden RTJM, Romein B, Heijnen JJ, Hellinga C, Luyben KCAM: Linear constraint relations in biochemical reaction systems: III. Sequential application of data reconciliation for sensitive detection of systematic errors. Biotechnol Bioeng 1994, 44:781–791. CrossRef
- Meyer V, Ram AFJ, Punt PJ: Genetics, genetic manipulation, and approaches to strain improvement of filamentous fungi. In Manual of Industrial Microbiology and Biotechnology. 3rd edition. Edited by: Baltz RH, Davies EJ, Demain AL. American Society for Microbiology, Washington DC; 2010:318–329.
- Ramachandran S, Fontanille P, Pandey A, Larroche C: Gluconic acid: properties, applications and microbial production. Food Technol Biotechnol 2006,44(2):185–195.
- Solomon PS, Waters ODC, Oliver RP: Decoding the mannitol enigma in filamentous fungi. Trends Microbiol 2007,15(6):257–262. CrossRef
- Sangeetha PT, Ramesh MN, Prapulla SG: Recent trends in the microbial production, analysis and application of Fructooligosaccharides. Trends Food Sci Technol 2005,16(10):442–457. CrossRef
- Posch AE, Spadiut O, Herwig C: A novel method for fast and statistically verified morphological characterization of filamentous fungi. Fungal Genet Biol
- Pirt SJ: A theory of the mode of growth of fungi in the form of pellets in submerged culture. Proc R Soc Lond B Biol Sci 1966,166(4):369–373. CrossRef
- Li X-B, Zhao G-R, Yuan Y-J: A strategy of phosphorus feeding for repeated fed-batch fermentation of penicillin G. Biochem Eng J 2005,27(1):53–58. CrossRef
- Nielsen J, Johansen CL, Villadsen J: Culture fluorescence measurements during batch and fed-batch cultivations with Penicillium chrysogenum. J Biotechnol 1994,38(1):51–62. CrossRef
- Ballio A, Russi S: The structure of a trisaccharide synthesized by action of Penicillium chrysogenum on sucrose. Gazz Chim Ital 1956, 86:476–483.
- De AFD, Russi S, Ballio A: Action of Penicillium chrysogenum on sucrose. Rend Ist Super Sanita (Ital Ed) 1955, 18:1065–1083.
- Justen P, Paul GC, Nienow AW, Thomas CR: Dependence of Penicillium chrysogenum growth, morphology, vacuolation, and productivity in fed-batch fermentations on impeller type and agitation intensity. Biotechnol Bioeng 1998,59(6):762–775. CrossRef
- Switching industrial production processes from complex to defined media: method development and case study using the example of Penicillium chrysogenum
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
Microbial Cell Factories
- Online Date
- June 2012
- Online ISSN
- BioMed Central
- Additional Links
- Filamentous fungi
- Complex media
- Defined media
- Stoichiometric mass balancing
- Fast strain characterization