Skip to main content
SpringerLink
Log in

High-throughput screening of high-yield colonies of Rhizopus oryzae for enhanced production of fumaric acid

  • Original Article
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

Fumaric acid is an important four-carbon dicarboxylic acid as a potential biorefinery target. A high-throughput screening method for fumaric acid overproduction strains was established. Nystatin (50 mg/L) was added into the production medium to restrict the spread of Rhizopus oryzae hyphae on agar plates. With bromocerol green as a pH indicator in the agar plates, the capability of fumaric acid biosynthesis by single colony was positively correlated with the diameter ratio of the colored ring and the colony. With this novel strategy, one high-yield mutant (Rhizopus oryzae ZJU11) was isolated from a large colony library of Rhizopus oryzae after UV irradiation. Starting with an optimized glucose concentration of 85 g/L, Rhizopus oryzae ZJU11 can produce 57.4 g/L fumaric acid in flask and 41.1 g/L in 5-L fermentor, which were 205% and 160% higher than that of the parent strain, respectively. Further studies showed that the production of fumaric acid by Rhizopus oryzae ZJU11 remained at the same level after three consecutive generations on the fermentation medium.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Cao NJ, Du JX, Chen CS, Gong CS, Tsao GT (1996a) Production of fumaric acid by immobilized Rhizopus using rotary biofilm contactor. Appl Biochem Biotechnol 63:387–394

    Article  Google Scholar 

  • Cao NJ, Du JX, Gong CS, Tsao GT (1996b) Simultaneous production and recovery of fumaric acid from immobilized Rhizopus oryzae with a rotary biofilm contactor and an adsorption column. Appl Environ Microbiol 62:2926–2931

    CAS  PubMed  Google Scholar 

  • Carta FS, Soccol CR, Ramos LP, Fontana JD (1999) Production of fumaric acid by fermentation of enzymatic hydrolysates derived from cassava bagasse. Bioresour Technol 68:23–28

    Article  CAS  Google Scholar 

  • Chen J, Chen HM, Zhu XC, Lu YH, Yang ST, Xu ZN, Cen PL (2009) Long-term production of soluble human Fas ligand through immobilization of Dictyostelium discoideum in a fibrous bed bioreactor. Appl Microbiol Biotechnol 82:241–248

    Article  CAS  PubMed  Google Scholar 

  • Engel CAR, Straathof AJJ, Zijlmans TW, van Gulik WM, van der Wielen LAM (2008) Fumaric acid production by fermentation. Appl Microbiol Biotechnol 78:379–389

    Article  Google Scholar 

  • Gangl IC, Weigand WA, Keller FA (1989) Economic comparison of calcium fumarate and sodium fumarate production by Rhizopus-Arrhizus. Appl Biochem Biotechnol 24:663–677

    Article  Google Scholar 

  • Goldberg I, Rokem JS, Pines O (2006) Organic acids: old metabolites, new themes. J Chem Technol Biotechnol 81:1601–1611

    Article  CAS  Google Scholar 

  • Jiang L, Wang JF, Liang SZ, Wang XN, Cen PL, Xu ZN (2009a) Production of butyric acid from glucose and xylose with immobilized cells of Clostridium tyrobutyricum in a fibrous-bed bioreactor. Appl Biochem Biotechnol 160:350–359

    Article  Google Scholar 

  • Jiang L, Wang JF, Liang SZ, Wang XN, Cen PL, Xu ZN (2009b) Butyric acid fermentation in a fibrous bed bioreactor with immobilized Clostridium tyrobutyricum from cane molasses. Bioresour Technol 100:3403–3409

    Article  CAS  PubMed  Google Scholar 

  • Kenealy W, Zaady E, Dupreez JC, Stieglitz B, Goldberg I (1986) Biochemical aspects of fumaric-acid accumulation by Rhizopus-Arrhizus. Appl Environ Microbiol 52:128–133

    CAS  PubMed  Google Scholar 

  • Liao W, Liu Y, Frear C, Chen SL (2007) A new approach of pellet formation of a filamentous fungus - Rhizopus oryzae. Bioresour Technol 98:3415–3423

    Article  CAS  PubMed  Google Scholar 

  • Moresi M, Parente E, Petruccioli M, Federici F (1991) Optimization of fumaric-acid production from potato flour by Rhizopus-Arrhizus. Appl Microbiol Biotechnol 36:35–39

    Article  CAS  Google Scholar 

  • Petruccioli M, Angiani E, Federici F (1996) Semi-continuous fumaric acid production by Rhizopus arrhizus immobilized in polyurethane sponge. Process Biochem 31:463–469

    Article  CAS  Google Scholar 

  • Rhodes RA, Moyer AJ, Smith ML, Kelley SE (1959) Production of fumaric acid by Rhizopus-Arrhizus. Appl Microbiol 7:74–80

    CAS  PubMed  Google Scholar 

  • Tao WY, Collier JR, Collier BJ, Negulescu II (1993) Fumaric-acid as an adhesion promoter in rayon nylon composite fibers. Tex Res J 63:162–170

    Article  CAS  Google Scholar 

  • Xu ZN, Shen WH, Chen XY, Lin JP, Cen PL (2005) A high-throughput method for screening of rapamycin-producing strains of Streptomyces hygroscopicus by cultivation in 96-well microtiter plates. Biotechnol Lett 27:1135–1140

    Article  CAS  PubMed  Google Scholar 

  • Xu ZN, Yang ST (2007) Production of mycophenolic acid by immobilized cells of Penicillium brevicompactum in a rotating fibrous bed bioreactor. Enzyme Microb Technol 40:623–628

    Article  CAS  Google Scholar 

  • Zhou Y, Du J, Tsao GT (2002) Comparison of fumaric acid production by Rhizopus oryzae using different neutralizing agents. Bioprocess Biosyst Eng 25:179–181

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Basic Research Program of China (2007CB707805), The Ministry of Science and Technology, China, and The National Science Foundation of China (20736008 and 20676115), China.

Author information

Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, 310027, People’s Republic of China

    Lei Huang, Ru Zang, Zhinan Xu & Peilin Cen

  2. School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, 310027, People’s Republic of China

    Peilian Wei

  3. Institute of Bioengineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, 310027, People’s Republic of China

    Zhinan Xu

Authors
  1. Lei Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peilian Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ru Zang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhinan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peilin Cen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhinan Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, L., Wei, P., Zang, R. et al. High-throughput screening of high-yield colonies of Rhizopus oryzae for enhanced production of fumaric acid. Ann Microbiol 60, 287–292 (2010). https://doi.org/10.1007/s13213-010-0039-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13213-010-0039-y

Keywords

Search

Navigation