Skip to main content
SpringerLink
Log in

Statistical Optimizing of Medium for Clavulanic Acid Production by Streptomyces clavuligerus Using Response Surface Methodology

  • Original Article
  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Clavulanic acid (CA) is a naturally occurring antibiotic produced by Streptomyces clavuligerus. Statistical optimization of the fermentation medium for CA production by Streptomyces clavuligerus was carried out. Multiple carbon sources, glycerol, dextrin, and triolein, were considered simultaneously. A two-level fractional factorial design experiment was conducted to identify the significant components of medium on CA production. Statistical analysis of the results showed that soybean meal, dextrin, and triolein were the most significant medium ingredients on CA production. The optimal level of these screened components was obtained by RSM based on the result of a Box-Behnken design, in which the values of dextrin, soybean meal, and triolein in CA fermentation medium were 12.37 g/L, 39.75 g/L, and 26.98 ml/L, respectively. Using the proposed optimized medium, the model predicted 938 mg/L of CA level and via experimental rechecking the model, 946 mg/L of CA level was attained in shake flask fermentation, significantly high than 630 mg/L of original medium. The optimized medium was further verified in 50-L stirred fermenter, and compared with performance of original medium in parallel, CA titer was increased from 889 to 1310 mg/L; a 47% increase was achieved through medium optimization by statistical approaches.

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

Data availability

Not applicable.

References

  1. Paradkar, A. (2013). Clavulanic acid production by Streptomyces clavuligerus: biogenesis, regulation and strain improvement. The Journal of Antibiotics, 66, 411–420.

    Article  CAS  Google Scholar 

  2. Huttner, A., Bielicki, J., Clements, M. N., Frimodt-Møller, N., Muller, A. E., Paccaud, J.-P., & Mouton, J. W. (2020). Oral amoxicillin and amoxicillin–clavulanic acid: Properties, indications and usage. Clinical Microbiology & Infection, 26(7), 871–879.

    Article  CAS  Google Scholar 

  3. Valentine, B. P., Bailey, C. R., Doherty, A., Morris, J., Elson, S. W., Bagley, K. H., & Nicholson, N. H. (1993). Evidence that arginine is later metabolic intermediate than ornithine in the biosynthesis of clavulanic acid by Streptomyces clavuligerus. Journal of the Chemical Society, Chemical Communications, 15, 1210–1211.

    Article  Google Scholar 

  4. Peng, Q., Gao, G., Lü, J., Long, Q., Chen, X., Zhang, F., Xu, M., Liu, K., Wang, Y., & Deng, Z. (2018). Engineered streptomyces lividans strains for optimal identification and expression of cryptic biosynthetic gene clusters. Frontiers in Microbiology, 9, 1–15.

    Article  Google Scholar 

  5. Song, J. Y., Jensen, S. E., & Lee, K. J. (2010). Clavulanic acid biosynthesis and genetic manipulation for its overproduction. Applied Microbiology and Biotechnology, 88, 659–669.

    Article  CAS  Google Scholar 

  6. Jnawali, H. N., Yoo, J. C., & Sohng, J. K. (2011). Improvement of clavulanic acid production in Streptomyces clavuligerus by genetic manipulation of structural biosynthesis genes. Biotechnology Letters, 33, 1221–1226.

    Article  CAS  Google Scholar 

  7. Kurt Kizildogan, A., VanliJaccard, G., Mutlu, A., Sertdemir, I., & Özcengiz, G. (2017). Genetic engineering of an industrial strain of Streptomyces clavuligerus for further enhancement of clavulanic acid production. Turkish Journal of Biology, 41, 342–353.

    Article  Google Scholar 

  8. Song, J. Y., Kim, E. S., Kim, D. W., Jensen, S. E., & Lee, K. J. (2008). Functional effects of increased copy number of the gene encoding proclavaminate amidino hydrolase on clavulanic acid production in Streptomyces clavuligerus ATCC 27064. Journal of Microbiology and Biotechnology, 18, 417–426.

    PubMed  Google Scholar 

  9. Saudagar, P. S., & Singhal, R. S. (2007). Optimization of nutritional requirements and feeding strategies for clavulanic acid production by Streptomyces clavuligerus. Bioresource Technology, 98, 2010–2017.

    Article  CAS  Google Scholar 

  10. Domingues, L. C. G., Teodoro, J. C., Hokka, C. O., Badino, A. C., & Araujo, M. L. G. C. (2010). Optimization of the glycerol-to-ornithine molar ratio in the feed medium for the continuous production of clavulanic acid by Streptomyces clavuligerus. Biochemical Engineering Journal, 53, 7–11.

    Article  CAS  Google Scholar 

  11. Chen, K. C., Lin, Y. H., Tsai, C. M., Hsieh, C. H., & Houng, J. Y. (2002). Optimization of glycerol feeding for clavulanic acid production by Streptomyces clavuligerus with glycerol feeding. Biotechnology Letters, 24, 455–458.

    Article  CAS  Google Scholar 

  12. Chen, K. C., Lin, Y. H., Wu, J. Y., & Hwang, S. C. (2003). Enhancement of clavulanic acid production in Streptomyces clavuligerus with ornithine feeding. Enzyme and Microbial Technology, 32, 152–156.

    Article  Google Scholar 

  13. Wang, Y. H., Yang, B., Ren, J., Dong, M. L., Liang, D., & Xu, A. L. (2005). Optimization of medium composition for the production of clavulanic acid by Streptomyces clavuligerus. Process Biochemistry, 40, 1161–1166.

    Article  CAS  Google Scholar 

  14. Ortiz, S. C., Hokka, C. O., & Badino, A. C. (2007). Utilization of soybean derivatives on clavulanic acid production by Streptomyces clavuligerus. Enzyme and Microbial Technology, 40, 1071–1077.

    Article  CAS  Google Scholar 

  15. Silva Rodrigues, K. C., de Souza, A. T., Badino, A. C., Pedrolli, D. B., & Cerri, M. O. (2018). Microbiology Screening of medium constituents for clavulanic acid production by Streptomyces clavuligerus. Brazilian Journal of Microbiology, 49, 832–839.

    Article  Google Scholar 

  16. Teodoro, J. C., Baptista-Neto, A., Cruz-Hernandez, I. L., Hokka, C. O., & Badino, A. C. (2006). Influence of feeding conditions on clavulanic acid production in fed-batch cultivation with medium containing glycerol. Applied Microbiology and Biotechnology, 72, 450–455.

    Article  CAS  Google Scholar 

  17. Large, K. P., Mirjalili, N., Osborne, M., Peacock, L. M., Zormpaidis, V., Walsh, M., Cavanagh, M. E., Leadlay, P. F., & Ison, A. P. (1999). Lipase activity in Streptomycetes. Enzyme and Microbial Technology, 25, 569–575.

  18. Young, T., Li, Y., & Efthimiou, G. (2019). Olive pomace oil can be used as an alternative carbon source for clavulanic acid production by Streptomyces clavuligerus. Waste and Biomass Valorization. https://doi.org/10.1007/s12649-019-00719-5.

  19. Maranesi, G. L., Baptista-Neto, A., Hokka, C. O., & Badino, A. C. (2005). Utilization of vegetable oil in the production of clavulanic acid by Streptomyces clavuligerus ATCC 27064. World Journal of Microbiology and Biotechnology, 21, 509–514.

    Article  CAS  Google Scholar 

  20. Salem-Bekhit, M. M., Alanazi, F. K., & Alsarra, I. A. (2010). Improvement and enhancement of clavulanic acid production in Streptomyces clavuligerus using vegetable oils. African Journal of Biotechnology, 9, 6806–6812.

    CAS  Google Scholar 

  21. Kim, S.-J., Kim, J.-O., Shin, C.-H., Park, H. W., & Kim, C. W. (2009). An approach to strain improvement and enhance production of clavulanic acid in Streptomyces clavuligerus. Bioscience, Biotechnology, and Biochemistry, 73(1), 160–164.

    Article  CAS  Google Scholar 

  22. Lee, S. D., Park, S. W., Oh, K. K., Hong, S. I., & Kim, S. W. (2002). Improvement for the production of clavulanic acid by mutant Streptomyces clavuligerus. Letters in Applied Microbiology, 34, 370–375.

    Article  Google Scholar 

  23. Teodoro, J. C., Araujo, M. L. G. C., Hokka, C. O., & Badino, A. C. (2010). Influence of glycerol and ornithine feeding on clavulanic acid production by Streptomyces clavuligerus. Brazilian Journal of Chemical Engineering, 27, 499–506.

  24. Ser, H.-L., Law, J.W.-F., Chaiyakunapruk, N., Jacob, S. A., Palanisamy, U. D., Chan, K.-G., Goh, B.-H., & Lee, L.-H. (2016). Fermentation conditions that affect clavulanic acid production in Streptomyces clavuligerus: A systematic review. Frontiers in Microbiology, 7, 522.

  25. Bellão, C., Antonio, T., Araujo, M. L. G. C., & Badino, A. C. (2013). Production of clavulanic acid and cephamycin c by streptomyces clavuligerus under different fed-batch conditions. Brazilian Journal of Chemical Engineering, 30, 257–266.

    Article  Google Scholar 

  26. Thakur, R., Roy, M. K., Dutta, N. N., & Bezbaruah, R. L. (1999). Coordinate production of cephamycin c and clavulanic acid by Streptomyces clavuligerus. Indian Journal of Experimental Biology, 37, 1031–1033.

Download references

Funding

This study was supported by the National Hi-Tech Program (863 Program 2006AA020302).

Author information

Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People’s Republic of China

    Tao Feng, Jie Zhao, Ju Chu, Yong-hong Wang & Ying-ping Zhuang

  2. Sinopharm Weiqida Pharmaceutical Co., Ltd, No.1 Medical Park, Datong Economic Development Zone, Datong, Shanxi Province, 037300, People’s Republic of China

    Tao Feng

Authors
  1. Tao Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jie Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ju Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yong-hong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ying-ping Zhuang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Tao Feng: experiment design, execution, and paper writing; Jie Zhao: experiment execution for fermentation by 50 L fermentor. Ju Chu is the corresponding author and guides the research direction of the project. Yong-hong Wang supports the data statistical analysis. Ying-Ping Zhuang supports experiment design and data analysis.

Corresponding author

Correspondence to Ju Chu.

Ethics declarations

Ethics Approval

Not applicable.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Competing Interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Feng, T., Zhao, J., Chu, J. et al. Statistical Optimizing of Medium for Clavulanic Acid Production by Streptomyces clavuligerus Using Response Surface Methodology. Appl Biochem Biotechnol 193, 3936–3948 (2021). https://doi.org/10.1007/s12010-021-03627-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-021-03627-4

Keywords

Search

Navigation