Combined available nitrogen resources enhanced erythromycin production and preliminary exploration of metabolic flux analysis under nitrogen perturbations

  • Qi Zhang
  • Haifeng Hang
  • Xiwei Tian
  • Wei Zeng
  • Zhenhua Yu
  • Xiaojian Wang
  • Yin Tang
  • Yingping ZhuangEmail author
  • Ju ChuEmail author
Research Paper


In the current study, the effect of different available nitrogen sources on erythromycin fermentation by Saccharopolyspora erythraea No. 8 is evaluated. Three different combinations of corn steep liquor and yeast powder were developed to investigate their impacts on erythromycin production. The results indicate that the optimal combination of available nitrogen sources was 10.0 g/L corn steep liquor and 4.0 g/L yeast power, generating a maximum yield of erythromycin of 13672 U/mL. To explore the effects of nitrogen perturbations on cell metabolism, metabolic flux analyses were performed and compared under different conditions. A high flux pentose phosphate pathway provided more NADPH for erythromycin synthesis via nitrogen optimization. Moreover, high n-propanol specific consumption rate enhanced erythromycin synthesis and n-propanol flowed into the central carbon metabolism by methylmalonyl-CoA node. These results indicate that the selection of an appropriate organic nitrogen source is essential for cell metabolism and erythromycin synthesis, and this is the first report of the successful application of available nitrogen source combinations in industrial erythromycin production.


Erythromycin Available nitrogen sources Corn steep liquor Yeast powder Metabolic flux 



This work was financially supported by the National Key Research and Development Program (2017YFB0309302), the National Basic Research Program of China (973 Program) (No. 2012CB721006), National Natural Science Foundation of China (No. 21276081) and the Fundamental Research Funds for the Central Universities (22221818014).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

449_2019_2171_MOESM1_ESM.doc (52 kb)
Biochemical reactions in the metabolic model (DOC 51 kb)


  1. 1.
    Wiley PF, Gerzon K, Flynn EH, Weaver O, Quarck UC, Chauvette RR, Monahan R (1957) Erythromycin. X1. Structure of erythromycin. J Am Chem Soc 79(22):6062–6070Google Scholar
  2. 2.
    Mironov VA, Sergienko OV, Nastasyak IN, Danilenko VN (2004) Biogenesis and Regulation of Biosynthesis of Erythromycins in Saccharopolyspora erythraea. Appl Biochem Microbiol 40(6):531–541Google Scholar
  3. 3.
    Taubman SB, Young FE, Corcoran JW (1963) Antibiotic glycosides, IV. studies on the mechanism of erythromycin resistance in Bacillus subtilis. Proc Natl Acad Sci USA 50(5):955–962Google Scholar
  4. 4.
    Taubman SB, Jones NR, Young FE, Corcoran JW (1966) Sensitivity and resistance to erythromycin in Bacillus subtilis 168: the ribosomal binding of erythromycin and chloramphenicol. Biochim Biophys Acta 123(2):438–440Google Scholar
  5. 5.
    Baltz RH (2006) Molecular engineering approaches to peptide, polyketide and other antibiotics. Nat Biotechnol 24(12):1533–1540Google Scholar
  6. 6.
    Chen CC, Hong M, Chu J, Huang MZ, Ouyang LM, Tian XW, Zhuang YP (2017) Blocking the flow of propionate into TCA cycle through a mutB knockout leads to a significant increase of erythromycin production by an industrial strain of Saccharopolyspora erythraea. Bioprocess Biosyst Eng 40(2):201–209Google Scholar
  7. 7.
    Wang Y, Wang YG, Chu J, Zhuang YP, Zhang LX, Zhang SL (2007) Improved production of erythromycin A by expression of a heterologous gene encoding S-adenosylmethionine synthetase. Appl Microbiol Biotechnol 75(4):837–842Google Scholar
  8. 8.
    Minas W, Brunker P, Kallio PT, Bailey JE (1998) Improved erythromycin production in a genetically engineered industrial strain of Saccharopolyspora erythraea. Biotechnol Prog 14(4):561–566Google Scholar
  9. 9.
    El-Enshasy HA, Mohamed NA, Farid MA, El-Diwany AI (2008) Improvement of erythromycin production by Saccharopolyspora erythraea in molasses based medium through cultivation medium optimization. Bioresour Technol 99(10):4263–4268Google Scholar
  10. 10.
    Devi CS, Saini A, Rastogi S, Naine SJ, Mohanasrinivasan V (2015) Strain improvement and optimization studies for enhanced production of erythromycin in bagasse based medium using Saccharopolyspora erythraea MTCC 1103. 3 Biotech 5(1):23–31Google Scholar
  11. 11.
    Zou X, Hang HF, Chen CF, Chu J, Zhuang YP, Zhang SL (2008) Application of oxygen uptake rate and response surface methodology for erythromycin production by Saccharopolyspora erythraea. J Ind Microbiol Biotechnol 35(12):1637–1642Google Scholar
  12. 12.
    Potvin J, Péringer P (1994) Ammonium regulation in Saccharopolyspora erythraea. Part I: Growth and antibiotic production. Biotechnol Lett 16(1):63–68Google Scholar
  13. 13.
    Martin JF, Demain AL (1980) Control of antibiotic biosynthesis. Microbiol Rev 44(2):230–251Google Scholar
  14. 14.
    Rafieenia R (2013) Effect of nutrients and culture conditions on antibiotic synthesis in Streptomycetes. Asian J Pharm Hea Sci 3(3):810–815Google Scholar
  15. 15.
    Zou X, Hang HF, Chu J, Zhuang YP, Zhang SL (2009) Enhancement of erythromycin A production with feeding available nitrogen sources in erythromycin biosynthesis phase. Bioresour Technol 100(13):3358–3365Google Scholar
  16. 16.
    Lara AR, Galindo E, Ramírez OT, Palomares LA (2006) Living with heterogeneities in bioreactors. Mol Biotechnol 34(3):355–381Google Scholar
  17. 17.
    Chen CF, Qi XC, Qian JC, Zhuang YP, Chu J, Zhang SL (2009) Effects of the dissolved oxygen on the erythromycin components of recombinant strain Saccharopolyspora erythraea ZLl004 fermentation. Chin J Antibio 34(11):659–663Google Scholar
  18. 18.
    Chen Y, Wang ZJ, Chu J, Zhuang YP, Zhang SL, Yu XG (2013) Significant decrease of broth viscosity and glucose consumption in erythromycin fermentation by dynamic regulation of ammonium sulfate and phosphate. Bioresour Technol 134(2):173–179Google Scholar
  19. 19.
    Chen Y, Huang MZ, Wang ZJ, Chu J, Zhuang YP, Zhang SL (2013) Controlling the feed rate of glucose and propanol for the enhancement of erythromycin production and exploration of propanol metabolism fate by quantitative metabolic flux analysis. Bioprocess Biosyst Eng 36(10):1445–1453Google Scholar
  20. 20.
    Zou X, Hang HF, Chu J, Zhuang YP, Zhang SL (2009) Oxygen uptake rate optimization with nitrogen regulation for erythromycin production and scale-up from 50 L to 372 m3 scale. Bioresour Technol 100(3):1406–1412Google Scholar
  21. 21.
    Zou X, Zeng W, Chen CF, Qi XC, Qian JC, Chu J, Zhuang YP, Zhang SL, Li WJ (2010) Fermentation optimization and industrialization of recombinant Saccharopolyspora erythraea strains for improved erythromycin a production. Biotechnol Bioprocess Eng 15(6):959–968Google Scholar
  22. 22.
    Zhao HT, Pang KY, Lin WL, Wang ZJ, Gao DQ, Guo MJ, Zhuang YP (2016) Optimization of the n-propanol concentration and feedback control strategy with electronic nose in erythromycin fermentation processes. Process Biochem 51(2):195–203Google Scholar
  23. 23.
    Maeda RN, Silva MMPD, Anna LMMS Jr (2010) Nitrogen source optimization for cellulase production by Penicillium funiculosum, using a sequential experimental design methodology and the desirability function. Appl Biochem Biotechnol 161(1–8):411–422Google Scholar
  24. 24.
    Corcoran JW (1981) Biochemical mechanisms in the biosynthesis of the erythromycins. Springer, BerlinGoogle Scholar
  25. 25.
    Reeves AR, Brikun IA, Cernota WH, Leach BI, Gonzalez MC, Weber JM (2007) Engineering of the methylmalonyl-CoA metabolite node for increased erythromycin production in oil-based fermentations of Saccharopolyspora erythraea. Metab Eng 9(3):293–303Google Scholar
  26. 26.
    Jung WS, Yoo YJ, Park JW, Park SR, Han AR, Ban YH, Kim EJ, Kim E, Yoon YJ (2011) A combined approach of classical mutagenesis and rational metabolic engineering improves rapamycin biosynthesis and provides insights into methylmalonyl-CoA precursor supply pathway in Streptomyces hygroscopicus ATCC 29253. Appl Microbiol Biotechnol 91(5):1389–1397Google Scholar
  27. 27.
    Li YY, Chang X, Yu WB, Li H, Ye ZQ, Yu H, Liu BH, Zhang Y, Zhang SL, Ye BC (2013) Systems perspectives on erythromycin biosynthesis by comparative genomic and transcriptomic analyses of S. erythraea E3 and NRRL23338 strains. BMC Genom 14(1):523Google Scholar
  28. 28.
    Reeves AR, Brikun IA, Cernota WH, Leach BI, Gonzalez MC, Weber JM (2006) Effects of methylmalonyl-CoA mutase gene knockouts on erythromycin production in carbohydrate-based and oil-based fermentations of Saccharopolyspora erythraea. J Ind Microbiol Biotechnol 33(7):600–609Google Scholar
  29. 29.
    Kumpfmüller J, Methling K, Fang L, Pfeifer BA, Lalk M, Schweder T (2016) Production of the polyketide 6-deoxyerythronolide B in the heterologous host Bacillus subtilis. Appl Microbiol Biotechnol 100(3):1209–1220Google Scholar
  30. 30.
    Karničar K, Drobnak I, Petek M, Magdevska V, Horvat J, Vidmar R, Baebler Š, Rotter A, Jamnik P, Fujs Š (2016) Integrated omics approaches provide strategies for rapid erythromycin yield increase in Saccharopolyspora erythraea. Microb Cell Fact 15(1):93Google Scholar
  31. 31.
    Lee WH, Chin YW, Han NS, Kim MD, Seo JH (2011) Enhanced production of GDP-L-fucose by overexpression of NADPH regenerator in recombinant Escherichia coli. Appl Microbiol Biotechnol 91(4):967–976Google Scholar
  32. 32.
    Ahmad I, Shim WY, Jeon WY, Yoon BH, Kim JH (2012) Enhancement of xylitol production in Candida tropicalis by co-expression of two genes involved in pentose phosphate pathway. Bioprocess Biosyst Eng 35(1–2):199–204Google Scholar
  33. 33.
    Potvin J, Péringer P (1993) Influence of N-propanol on growth and antibiotic production by an industrial strain of Streptomyces erythreus under different nutritional conditions. Biotechnol Lett 15(5):455–460Google Scholar
  34. 34.
    Guo Q, Chu J, Zhuang YP, Gao Y (2016) Controlling the feed rate of propanol to optimize erythromycin fermentation by on-line capacitance and oxygen uptake rate measurement. Bioprocess Biosyst Eng 39(2):255–265Google Scholar
  35. 35.
    Chen Y, Wang Z, Chu J, Xi B, Zhuang Y (2015) The glucose RQ-feedback control leading to improved erythromycin production by a recombinant strain Saccharopolyspora erythraea ZL1004 and its scale-up to 372-m3 fermenter. Bioprocess Biosyst Eng 38(1):105–112Google Scholar
  36. 36.
    Chen Y, Deng W, Wu JQ, Qian JC, Chu J, Zhuang YP, Zhang SL, Liu W (2008) Genetic modulation of the overexpression of tailoring genes eryK and eryG leading to the improvement of erythromycin a purity and production in Saccharopolyspora erythraea fermentation. Appl Environ Microbiol 74(6):1820–1828Google Scholar
  37. 37.
    Yao LL, Liao CH, Huang G, Zhou Y, Rigali S, Zhang BC, Ye BC (2014) GlnR-mediated regulation of nitrogen metabolism in the actinomycete Saccharopolyspora erythraea. Appl Microbiol Biotechnol 98(18):7935–7948Google Scholar
  38. 38.
    Liao CH, Yao L, Xu Y, Liu WB, Zhou Y, Ye BC (2015) Nitrogen regulator GlnR controls uptake and utilization of non-phosphotransferase-system carbon sources in actinomycetes. Proc Natl Acad Sci USA 112(51):15630–15635Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Qi Zhang
    • 1
  • Haifeng Hang
    • 1
  • Xiwei Tian
    • 1
  • Wei Zeng
    • 2
  • Zhenhua Yu
    • 2
  • Xiaojian Wang
    • 2
  • Yin Tang
    • 1
  • Yingping Zhuang
    • 1
    Email author
  • Ju Chu
    • 1
    Email author
  1. 1.State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiPeople’s Republic of China
  2. 2.Yidu HEC Biochem. Co. Ltd.YiduPeople’s Republic of China

Personalised recommendations