Skip to main content
SpringerLink
Log in

Metabolic control analysis of l-lactate synthesis pathway in Rhizopus oryzae As 3.2686

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

The relationship between the metabolic flux and the activities of the pyruvate branching enzymes of Rhizopus oryzae As 3.2686 during l-lactate fermentation was investigated using the perturbation method of aeration. The control coefficients for five enzymes, pyruvate dehydrogenase (PDH), pyruvate carboxylase (PC), pyruvate decarboxylase (PDC), lactate dehydrogenase (LDH), and alcohol dehydrogenase (ADH), were calculated. Our results indicated significant correlations between PDH and PC, PDC and LDH, PDC and ADH, LDH and ADH, and PDC and PC. It also appeared that PDH, PC, and LDH strongly controlled the l-lactate flux; PDH and ADH strongly controlled the ethanol flux; while PDH and PC strongly controlled the acetyl coenzyme A flux and the oxaloacetate flux. Further, the flux control coefficient curves indicated that the control of the system gradually transferred from PDC to PC during the steady state. Therefore, PC was the key rate-limiting enzyme that controls the flux distribution.

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

Similar content being viewed by others

References

  1. Huang LP, Jin B, Lant P (2005) Direct fermentation of potato starch wastewater to lactic acid by Rhizopus oryzae and Rhizopus arrhizus. Bioprocess Biosyst Eng 27:229–238

    Article  Google Scholar 

  2. Thongchul N, Navankasattusas S, Yang ST (2010) Production of lactic acid and ethanol by Rhizopus oryzae integrated with cassava pulp hydrolysis. Bioprocess Biosyst Eng 33:407–416

    Article  CAS  Google Scholar 

  3. Chotisubha-anandha N, Thitiprasert S, Tolieng V, Thongchul N (2011) Improved oxygen transfer and increased L-lactic acid production by morphology control of Rhizopus oryzae in a static bed bioreactor. Bioprocess Biosyst Eng 34:163–172

    Article  CAS  Google Scholar 

  4. Phrueksawan P, Kulpreecha S, Sooksai S, Thongchul N (2012) Direct fermentation of L (+)-lactic acid from cassava pulp by solid state culture of Rhizopus oryzae. Bioprocess Biosyst Eng 35:1429–1436

    Article  CAS  Google Scholar 

  5. Stephanopoulos GN, Aristos AA, Nielsen J (1998) Metabolic engineering: principles and methodologies. Academic Press, San Diego

    Google Scholar 

  6. Nagamori E, Shimizu K, Fujita H, Tokuhiro K, Ishida N, Takahashi H (2013) Metabolic flux analysis of genetically engineered Saccharomyces cerevisiae that produces lactate under micro-aerobic conditions. Bioprocess Biosyst Eng 36:1261–1265

    Article  CAS  Google Scholar 

  7. Pitkänen JP, Aristidou A, Salusjärvi L, Ruohonen L, Penttilä M (2003) Metabolic flux analysis of xylose metabolism in recombinant Saccharomyces cerevisiae using continuous culture. Metab Eng 5:16–31

    Article  Google Scholar 

  8. Bai DM, Zhao XM, Li XG, Xu SM (2004) Strain improvement of Rhizopus oryzae for over-production of L(+)-lactic acid and metabolic flux analysis of mutants. Biochem Eng J 18:41–48

    Article  CAS  Google Scholar 

  9. Heijnen JJ, van Gulik WM, Shimizu H, Stephanopoulos G (2004) Metabolic flux control analysis of branch points: an improved approach to obtain flux control coefficients from large perturbation data. Metab Eng 6:391–400

    Article  CAS  Google Scholar 

  10. Conradie R, Westerhoff HV, Rohwer JM, Hofmeyr J-HS, Snoep JL (2006) Summation theorems for flux and concentration control coefficients of dynamic systems. IEE Proc Syst Biol 153:314–317

    Article  CAS  Google Scholar 

  11. Kurata H, Zhao QY, Okuda R, Shimizu K (2007) Integration of enzyme activities into metabolic flux distributions by elementary mode analysis. BMC Syst Biol 1:31

    Article  Google Scholar 

  12. Ouattara DA, Prot JM, Bunescu A, Dumas ME, Elena-Herrmann B, Leclerc E, Brochot C (2012) Metabolomics-on-a-chip and metabolic flux analysis for label-free modeling of the internal metabolism of HepG2/C3A cells. Mol BioSyst 8:1908–1920

    Article  CAS  Google Scholar 

  13. Nielsen J, Jørgensen HS (1995) Metabolic control analysis of the penicillin biosynthetic pathway in a high-yielding strain of Penicillium chrysogenum. Biotechnol Prog 11:299–305

    Article  CAS  Google Scholar 

  14. Shiraishi F, Sriyudthsak K, Suzuki Y (2010) Calculation errors of time-varying flux control coefficients obtained from elasticity coefficients by means of summation and connectivity theorems in metabolic control analysis. Math Biosci 223:105–114

    Article  Google Scholar 

  15. Preller A, Quiroga D, Wilson CAM, Ureta T (2007) In vivo determination of the flux control coefficients of the enzymes involved in glycogen synthesis. FEBS J 274:243

    Google Scholar 

  16. Ureta T, Preller A, Wilson C (2008) In vivo flux control coefficient of glycogen synthase in frog oocytes. FEBS J 275:435

    Article  Google Scholar 

  17. Sarıyar B, Perk S, Akman U, Hortaçsu A (2006) Monte Carlo sampling and principal component analysis of flux distributions yield topological and modular information on metabolic networks. J Theoret Biol 242:389–400

    Article  Google Scholar 

  18. Quarato G, Piccoli C, Scrima R, Capitanio N (2011) Variation of flux control coefficient of cytochrome c oxidase and of the other respiratory chain complexes at different values of protonmotive force occurs by a threshold mechanism. BBA Bioenerg 1807:1114–1124

    Article  CAS  Google Scholar 

  19. Salusjärvi L, Poutanen M, Pitkänen JP, Koivistoinen H, Aristidou A, Kalkkinen N, Ruohonen L, Penttilä M (2003) Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. Yeast 20:295–314

    Article  Google Scholar 

  20. Li XF, Qu RN (2012) Parameter identification and terminal steady-state optimization for S system in microbial continuous fermentation. Int J Biomath 5:73–83

    Google Scholar 

  21. Hashemi M, Mousavi SM, Razavi SH, Shojaosadati SA (2011) Mathematical modeling of biomass and α-amylase production kinetics by Bacillus sp. in solid-state fermentation based on solid dry weight variation. Biochem Eng J 53:159–164

    Article  CAS  Google Scholar 

  22. Zhong S, Qiu YF, Han BB, Zhao JY, Zhu XJ, Chen XX (2011) Detection of serum desmoglein antibody level using enzyme-linked immunosorbent assay (ELISA) for monitoring disease activity in patients with pemphigus vulgaris. Beijing Da Xue Xue Bao 43:14–415

    Google Scholar 

  23. Duan XZ, Wang M, Li HW, Zhuang H, Xu DP, Wang FS (2004) Decreased frequency and function of circulating plasmocytoid dendritic cells (pDC) in hepatitis B virus infected humans. J Clin Immunol 24:637–646

    Article  CAS  Google Scholar 

  24. Harb J, Lara I, Saleh O, Streif J, Khraiwesh B (2011) Treatments that suppress ethylene production or ethylene action modify ADH and AAT gene expression and aroma-related enzyme activities in ‘Delbarde Estivale’ apple: consequences for the aroma profiles of fruit. J Hortic Sci Biotech 86:182–188

    CAS  Google Scholar 

  25. Hu RF, Lin L, Liu TJ, Ouyang PK, He BH, Liu SJ (2008) Reducing sugar content in hemicellulose hydrolysate by DNS method: a revisit. J Biobased Mater Bioenerg 2:156–161

    Article  Google Scholar 

  26. Ke W, Zheng Z, Jiang ST, Luo SZ, Wu XF, Yang PZ (2014) The analysis of metabolic flux and enzyme activities of pyruvate branch point of Rhizopus oryzae As 3.2686. J Chem Eng Chin Univ 28:1071–1079 (in Chinese)

    CAS  Google Scholar 

  27. Liu RX, Kuang J, Gong Q, Hou XL (2003) Principal component regression analysis with SPSS. Comput Methods Programs Biomed 71:141–147

    Article  CAS  Google Scholar 

  28. Bai DM, Fu WM, Zhao XM, Dai HX, Li XG, Xu SM (2002) Optimization of L(+)-Lactic acid fermentation by Rhizopus oryzae R1021 by metabolic flux analysis. J Wuxi Univ Light Ind 21:554–558 (in Chinese)

    CAS  Google Scholar 

  29. Skory CD (2004) Lactic acid production by Rhizopus oryzae transformants with modified lactate dehydrogenase activity. Appl Microbiol Biot 64:237–242

    Article  CAS  Google Scholar 

  30. Lin H, Vadali RV, Bennett GN, San KY (2004) Increasing the acetyl-CoA pool in the presence of overexpressed phosphoenolpyruvate carboxylase or pyruvate carboxylase enhances succinate production in Escherichia coli. Biotechnol Prog 20:1599–1604

    Article  CAS  Google Scholar 

  31. Wu SG, Zhang T, Deng L (2011) Effect of over-expressing of pyruvate carboxylase gene on production of fumarate. Biotechnol Bull 12:175–180 (in Chinese)

    Google Scholar 

Download references

Acknowledgments

This work was funded by the National Natural Science Foundation of China (No. 31071636/31171741) and the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (No. 2010JYLH0837).

Author information

Author notes

    Authors and Affiliations

    1. School of Biotechnology and Food Engineering, Hefei University of Technology, 193 Tunxi Road, Hefei, 230009, People’s Republic of China

      Wei Ke, Shu Chang, Xiaoju Chen, Shuizhong Luo, Shaotong Jiang, Peizhou Yang, Xuefeng Wu & Zhi Zheng

    2. Key Laboratory for Agricultural Products Processing of Anhui Province, Hefei University of Technology, 193 Tunxi Road, Hefei, 230009, People’s Republic of China

      Shuizhong Luo, Shaotong Jiang, Peizhou Yang, Xuefeng Wu & Zhi Zheng

    Authors
    1. Wei Ke
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Shu Chang
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Xiaoju Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Shuizhong Luo
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Shaotong Jiang
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Peizhou Yang
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Xuefeng Wu
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Zhi Zheng
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Zhi Zheng.

    Additional information

    W. Ke and S. Chang contributed equally to this work.

    Appendix

    Appendix

    See Fig. 3 and Table 4.

    Fig. 3
    figure 3

    Metabolic network of Rhizopus oryzae As 3.2686 during growth on glucose

    Full size image
    Table 4 Metabolic nodes and flux equations of Rhizopus oryzae As 3.2686 during growth on glucose
    Full size table

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Ke, W., Chang, S., Chen, X. et al. Metabolic control analysis of l-lactate synthesis pathway in Rhizopus oryzae As 3.2686. Bioprocess Biosyst Eng 38, 2189–2199 (2015). https://doi.org/10.1007/s00449-015-1458-8

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00449-015-1458-8

    Keywords

    Search

    Navigation