Bioprocess and Biosystems Engineering

, Volume 37, Issue 6, pp 1133–1140 | Cite as

Production of alginate by Azotobacter vinelandii grown at two bioreactor scales under oxygen-limited conditions

  • Alvaro Díaz-BarreraEmail author
  • Javiera Gutierrez
  • Fabiola Martínez
  • Claudia Altamirano
Original Paper


The oxygen transfer rate (OTR) was evaluated as a scale-up criterion for alginate production in 3- and 14-L stirred fermentors. Batch cultures were performed at different agitation rates (200, 300, and 600 rpm) and airflow rates (0.25, 0.5, and 1 vvm), resulting in different maximum OTR levels (OTRmax). Although the two reactors had a similar OTRmax (19 mmol L−1 h−1) and produced the same alginate concentration (3.8 g L−1), during the cell growth period the maximum molecular weight of the alginate was 1,250 kDa in the 3-L stirred fermentor and 590 kDa in 14-L stirred fermentor. The results showed for the first time the evolution of the molecular weight of alginate and OTR profiles for two different scales of stirred fermentors. There was a different maximum specific oxygen uptake rate between the two fermenters, reaching 8.3 mmol g−1 h−1 in 3-L bioreactor and 10.6 mmol g−1 h−1 in 14-L bioreactor, which could explain the different molecular weights observed. These findings open the possibility of using \( q_{{{\text{O}}_{ 2} }} \) instead of OTRmax as a scaling criterion to produce polymers with similar molecular weights during fermentation.


Scale-up Alginate Molecular weight Oxygen transfer rate Stirred fermentor 



The authors are grateful to Fondecyt Grant 11110311 from CONICYT (Chile) for financial support.


  1. 1.
    Díaz-Barrera A, Soto E (2010) Biotechnological uses of Azotobacter vinelandii: current state, limits, and prospects. Af J Biotechnol 9:5240–5250Google Scholar
  2. 2.
    Hay I, Rehman Z, Ghafoor A, Rehm B (2010) Bacterial biosynthesis of alginates. J Chem Technol Biotechnol 85:752–759CrossRefGoogle Scholar
  3. 3.
    Peña C, Trujillo-Roldán M, Galindo E (2000) Influence of dissolved oxygen tension and agitation speed on alginate production and its molecular weight in cultures of Azotobacter vinelandii. Enzyme Microb Technol 27:390–398CrossRefGoogle Scholar
  4. 4.
    Peña C, Galindo E, Büchs J (2011) The viscosifying power, degree of acetylation and molecular mass of the alginate produced by Azotobacter vinelandii in shake flasks are determined by the oxygen transfer rate. Process Biochem 46:290–297CrossRefGoogle Scholar
  5. 5.
    Díaz-Barrera A, Peña C, Galindo E (2007) The oxygen transfer rate influences the molecular weight of the alginate produced by Azotobacter vinelandii. Appl Microbiol Biotechnol 76:903–910CrossRefGoogle Scholar
  6. 6.
    Lozano E, Galindo E, Peña C (2011) Oxygen transfer rate during the production of alginate by Azotobacter vinelandii under oxygen-limited and non oxygen-limited conditions. Microb Cell Fact 10:13CrossRefGoogle Scholar
  7. 7.
    Reyes C, Peña C, Galindo E (2003) Reproducing shake flasks performance in stirred fermentors: production of alginates by Azotobacter vinelandii. J Biotechnol 105:189–198CrossRefGoogle Scholar
  8. 8.
    Peña C, Millán M, Galindo E (2008) Production of alginate by Azotobacter vinelandii in a stirred fermentor simulating the evolution of power input observed in shake flasks. Process Biochem 43:775–778CrossRefGoogle Scholar
  9. 9.
    Puliga SL, Handa S, Gummadi SN, Doble M (2010) Enhancement and scale-up of β-(1,3) glucan production by Agrobacterium sp. Int J Food Eng 6:1CrossRefGoogle Scholar
  10. 10.
    Miller G (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal Chem 31:426–428CrossRefGoogle Scholar
  11. 11.
    Kayser A, Weber J, Hecht V, Rinas U (2005) Metabolic flux analysis of Escherichia coli in glucose-limited continuous culture I. Growth-rate dependent metabolic efficiency at steady state. Microbiology 151:693–706CrossRefGoogle Scholar
  12. 12.
    Maier U, Büchs J (2001) Characterisation of the gas–liquid mass transfer in shaking bioreactors. Biochem Eng J 7:99–106CrossRefGoogle Scholar
  13. 13.
    Oelze J (2000) Respiratory protection of nitrogenase in Azotobacter species: is a widely held hypothesis unequivocally supported by experimental evidence? FEMS Microbiol Rev 24:321–333CrossRefGoogle Scholar
  14. 14.
    Anderlei T, Zang W, Papaspyrou M, Büchs J (2004) Online respiration activity measurement (OTR, CTR, RQ) in shake flasks. Biochem Eng J 17:187–194CrossRefGoogle Scholar
  15. 15.
    Peña C, Peter C, Büchs J, Galindo E (2007) Evolution of the specific power consumption and oxygen transfer rate in alginate-producing cultures of Azotobacter vinelandii conducted in shake flasks. Biochem Eng J 36:73–80CrossRefGoogle Scholar
  16. 16.
    Kuhla J, Oelze J (1988) Dependency of growth yield, maintenance and K s-values on the dissolved oxygen concentration in continuous cultures of Azotobacter vinelandii. Arch Microbiol 149:509–514CrossRefGoogle Scholar
  17. 17.
    Garcia-Ochoa F, Gomez E (2009) Bioreactor scale-up and oxygen transfer rate in microbial processes: an overview. Biotechnol Adv 27:153–176CrossRefGoogle Scholar
  18. 18.
    Trujillo-Roldán M, Moreno S, Espín G, Galindo E (2004) The roles of oxygen and alginate-lyase in determining the molecular weight of alginate produced by Azotobacter vinelandii. Appl Microbiol Biotechnol 63:742–747CrossRefGoogle Scholar
  19. 19.
    Díaz-Barrera A, Aguirre A, Berrios J, Acevedo F (2011) Continuous cultures for alginate production by Azotobacter vinelandii growing at different oxygen uptake rates. Process Biochem 46:1879–1883CrossRefGoogle Scholar
  20. 20.
    Remminghorst U, Rehm B (2006) Bacterial alginates: from biosynthesis to applications. Biotechnol Lett 28:1701–1712CrossRefGoogle Scholar
  21. 21.
    Castillo T, Heinzle E, Peifer S, Schneider K, Peña C (2013) Oxygen supply strongly influences metabolic fluxes, the production of poly(3-hydroxybutyrate) and alginate, and the degree of acetylation of alginate in Azotobacter vinelandii. Process Biochem 48:995–1003CrossRefGoogle Scholar
  22. 22.
    Díaz-Barrera A, Silva P, Berrios J, Acevedo F (2010) Manipulating the molecular weight of the alginate produced by Azotobacter vinelandii in continuous cultures. Bioresour Technol 101:9405–9408CrossRefGoogle Scholar
  23. 23.
    Díaz-Barrera A, Soto E, Altamirano C (2012) Alginate production and alg8 gene expression by Azotobacter vinelandii in continuous cultures. J Ind Microbiol Biotechnol 39:613–621CrossRefGoogle Scholar
  24. 24.
    Wang ZJ, Wang HY, Li YL, Chu J, Huang MZ, Zhuang YP, Zhang Si-L (2010) Improved vitamin B12 production by step-wise reduction of oxygen uptake rate under dissolved oxygen limiting level during fermentation process. Bioresour Technol 101:2845–2852CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Alvaro Díaz-Barrera
    • 1
    Email author
  • Javiera Gutierrez
    • 1
  • Fabiola Martínez
    • 1
  • Claudia Altamirano
    • 1
    • 2
  1. 1.Escuela de Ingeniería BioquímicaPontificia Universidad Católica de ValparaísoCasillaChile
  2. 2.CREAS CONICYT-REGIONALValparaísoChile

Personalised recommendations