Advertisement

Bioprocess and Biosystems Engineering

, Volume 41, Issue 8, pp 1143–1151 | Cite as

Metabolic analyses of the improved ε-poly-l-lysine productivity using a glucose–glycerol mixed carbon source in chemostat cultures

  • Jian-Hua Zhang
  • Xin Zeng
  • Xu-Sheng Chen
  • Zhong-Gui Mao
Research Paper
  • 44 Downloads

Abstract

The glucose–glycerol mixed carbon source remarkably reduced the batch fermentation time of ε-poly-l-lysine (ε-PL) production, leading to higher productivity of both biomass and ε-PL, which was of great significance in industrial microbial fermentation. Our previous study confirmed the positive influence of fast cell growth on the ε-PL biosynthesis, while the direct influence of mixed carbon source on ε-PL production was still unknown. In this work, chemostat culture was employed to study the capacity of ε-PL biosynthesis in different carbon sources at a same dilution rate of 0.05 h−1. The results indicated that the mixed carbon source could enhance the ε-PL productivity besides the rapid cell growth. Analysis of key enzymes demonstrated that the activities of phosphoenolpyruvate carboxylase, citrate synthase, aspartokinase and ε-PL synthetase were all increased in chemostat culture with the mixed carbon source. In addition, the carbon fluxes were also improved in the mixed carbon source in terms of tricarboxylic acid cycle, anaplerotic and diaminopimelate pathway. Moreover, the mixed carbon source also accelerated the energy metabolism, leading to higher levels of energy charge and NADH/NAD+ ratio. The overall improvements of primary metabolism in chemostat culture with glucose–glycerol combination provided sufficient carbon skeletons and ATP for ε-PL biosynthesis. Therefore, the significantly higher ε-PL productivity in the mixed carbon source was a combined effect of both superior substrate group and rapid cell growth.

Keywords

ε-Poly-l-lysine Mixed carbon source Chemostat culture Metabolic analyses 

Notes

Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (31671846, 31301556), the Science and Technology Department of Jiangsu Province (BY2016022-25), the Fundamental Research Funds for the Central Universities (JUSRP51504), the Open Project Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (KLIBKF201302), and the Jiangsu Province Collaborative Innovation Center for Advanced Industrial Fermentation Industry Development Program.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

449_2018_1943_MOESM1_ESM.doc (1.2 mb)
Supplementary material 1 (DOC 1220 KB)

References

  1. 1.
    Hiraki J, Ichikawa T, Ninomiya S, Seki H, Uohama K, Seki H, Kimura S, Yanagimoto Y, Barnett JJ (2003) Use of ADME studies to confirm the safety of ε-polylysine as a preservative in food. Regul Toxicol Pharm 37:328–340CrossRefGoogle Scholar
  2. 2.
    Shih IL, Shen MH, Van YT (2006) Microbial synthesis of poly(epsilon–lysine) and its various applications. Bioresour Technol 97(9):1148–1159CrossRefGoogle Scholar
  3. 3.
    Bankar SB, Singhal RS (2013) Panorama of poly-ε-lysine. RSC Adv 3(23):8586–8603CrossRefGoogle Scholar
  4. 4.
    Li S, Tang L, Chen X, Liao L, Li F, Mao Z (2011) Isolation and characterization of a novel epsilon-poly-l-lysine producing strain: Streptomyces griseofuscus. J Ind Microbiol Biot 38(4):557–563CrossRefGoogle Scholar
  5. 5.
    Jia S, Wang G, Sun Y, Tan Z (2009) Improvement of ε-poly-l-lysine production by Streptomyces albulus TUST2 employing a feeding strategy. Paper presented at the Tianjin University of Science and Technology, Tianjin, ChinaGoogle Scholar
  6. 6.
    Bankar SB, Singhal RS (2010) Optimization of poly-epsilon-lysine production by Streptomyces noursei NRRL 5126. Bioresour Technol 101(21):8370–8375CrossRefGoogle Scholar
  7. 7.
    Chen XS, Tang L, Li S, Liao LJ, Zhang JH, Mao ZG (2011) Optimization of medium for enhancement of epsilon-poly-l-lysine production by Streptomyces sp. M-Z18 with glycerol as carbon source. Bioresour Technol 102(2):1727–1732CrossRefGoogle Scholar
  8. 8.
    Kahar P, Iwata T, Hiraki J, Park EY, Okabe M (2001) Enhancement of ε-polylysine production by Streptomyces albulus strain 410 using pH control. J Biosci Bioeng 91(2):190–194CrossRefGoogle Scholar
  9. 9.
    Chen XS, Ren XD, Dong N, Li S, Li F, Zhao FL, Tang L, Zhang JH, Mao ZG (2012) Culture medium containing glucose and glycerol as a mixed carbon source improves ε-poly-l-lysine production by Streptomyces sp. M-Z18. Bioproc Biosyst Eng 35(3):469–475CrossRefGoogle Scholar
  10. 10.
    Liu Y, Zhang YG, Zhang RB, Zhang F, Zhu J (2011) Glycerol/glucose co-fermentation: one more proficient process to produce propionic acid by Propionibacterium acidipropionici. Curr Microbiol 62(1):152–158CrossRefGoogle Scholar
  11. 11.
    Kim YS, Lee JH, Kim NH, Yeom SJ, Kim SW, Oh DK (2011) Increase of lycopene production by supplementing auxiliary carbon sources in metabolically engineered Escherichia coli. Appl Microbiol Biot 90(2):489–497CrossRefGoogle Scholar
  12. 12.
    Peacock L, Ward J, Ratledge C, Dickinson F, Ison A (2003) How Streptomyces lividans uses oils and sugars as mixed substrates? Enzyme Microb Tech 1:157–166CrossRefGoogle Scholar
  13. 13.
    Zeng X, Chen XS, Ren XD, Liu QR, Wang L, Sun QX, Tang L, Mao ZG (2014) Insights into the role of glucose and glycerol as a mixed carbon source in the improvement of ε-poly-l-lysine productivity. Appl Biochem Biotech 173(8):2211–2224CrossRefGoogle Scholar
  14. 14.
    Zeng X, Chen X, Ren X, Wang L, Gao Y, Mao Z (2016) Improved ε-poly-l-lysine productivity partly resulting from rapid cell growth in cultures using a glucose–glycerol mixed carbon source. Eng Life Sci 16:443–452CrossRefGoogle Scholar
  15. 15.
    Vázquez-Lima F, Silva P, Barreiro A, Martínez-Moreno R, Morales P, Quirós M, González R, Albiol J, Ferrer P (2014) Use of chemostat cultures mimicking different phases of wine fermentations as a tool for quantitative physiological analysis. Microb Cell Fact 13:85CrossRefGoogle Scholar
  16. 16.
    Gallmetzer M, Burgstaller W (2001) Citrate efflux in glucose-limited and glucose-sufficient chemostat culture of Penicillium simplicissium. Antonie Van Leeuwenhoek 79(1):81–87CrossRefGoogle Scholar
  17. 17.
    Pim VH, Johannes PVD, Jack TP (1998) Effect of specific growth rate on fermentative capacity of baker’s yeast. Appl Environ Microb 64:4226–4233Google Scholar
  18. 18.
    Nishikawa MOK (2002) Distribution of microbes producing antimicrobial ε-poly-l-lysine polymers in soil microflora determined by a novel method. Appl Environ Microb 68(7):3575–3581CrossRefGoogle Scholar
  19. 19.
    Fountoulakis M, Lahm HW (1998) Hydrolysis and amino acid composition analysis of proteins. J Chromatogr A 826(2):109–134CrossRefGoogle Scholar
  20. 20.
    Zhu Y, Rinzema A, Bonarius H, Tramper J, Bol J (1998) Microbial transglutaminase production by Streptoverticillium mobaraense: analysis of amino acid metabolism using mass balances. Enzyme Microb Tech 23(3):216–226CrossRefGoogle Scholar
  21. 21.
    Chen XS, Mao ZG (2013) Comparison of glucose and glycerol as carbon sources for epsilon-poly-l-lysine production by Streptomyces sp. M-Z18. Appl Biochem Biotech 170(1):185–197CrossRefGoogle Scholar
  22. 22.
    Yamanaka K, Maruyama C, Takagi H, Hamano Y (2008) Epsilon-poly-l-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase. Nat Chem Biol 4(12):766–772CrossRefGoogle Scholar
  23. 23.
    Yamanaka K, Kito N, Imokawa Y, Maruyama C, Utagawa T, Hamano Y (2010) Mechanism of epsilon-poly-l-lysine production and accumulation revealed by identification and analysis of an epsilon-poly-l-lysine-degrading enzyme. Appl Environ Microb 76(17):5669–5675CrossRefGoogle Scholar
  24. 24.
    VanBriesen JM (2002) Evaluation of methods to predict bacterial yield using thermodynamics. Biodegradation 13:171–190CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jian-Hua Zhang
    • 1
  • Xin Zeng
    • 2
  • Xu-Sheng Chen
    • 1
  • Zhong-Gui Mao
    • 1
  1. 1.The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of BiotechnologyJiangnan UniversityWuxiChina
  2. 2.College of Life SciencesHuaibei Normal UniversityHuaibeiChina

Personalised recommendations