Bioprocess and Biosystems Engineering

, Volume 40, Issue 2, pp 201–209 | Cite as

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

  • Chongchong Chen
  • Ming Hong
  • Ju Chu
  • Mingzhi Huang
  • Liming Ouyang
  • Xiwei Tian
  • Yingping Zhuang
Original Paper


A high erythromycin producing mutant strain Saccharopolyspora erythraea HL3168 E3-ΔmutB was constructed by deleting mutB (SACE_5639) gene encoding the beta subunit of methylmalonyl-CoA mutase of an industrial strain of S. erythraea HL3168 E3. Industrial media and process control strategies were adopted in a 5 L bioreactor for characterizing the physiological parameters. The total erythromycin titer and erythromycin A concentration in mutant were 46.9 (12740.5 μg/mL) and 64.9 % (8094.4 μg/mL) higher than those in original strain, respectively, which were comparable to industrial erythromycin production. The specific glucose and n-propanol consumption rates were increased by 52.4 and 39.8 %, respectively. During the rapid erythromycin synthesis phase, the yield of erythromycin on n-propanol also increased from 24.3 % in control group to 66.9 % in mutant group. Meanwhile, the specific formation rates of methylmalonyl-CoA and propionyl-CoA, two crucial precursors for erythromycin synthesis, were 1.89- and 2.02-folds higher in the mutant strain, respectively.


Saccharopolyspora erythraea Erythromycin N-propanol Methylmalonyl-CoA mutase mutB 



This work was financially supported by a Grant from the Major State Basic Research Development Program of China (973 Program, No. 2012CB721006) and National Natural Science Foundation of China (No. 21276081).

Compliance with ethical standards

Conflict of interest

The authors declare no financial or commercial conflict of interest.

Supplementary material

449_2016_1687_MOESM1_ESM.doc (203 kb)
Supplementary material 1 (DOC 203 kb)


  1. 1.
    Isaac OK, Jos H, Eugene R, Hubert V, Ludo V, Maurice D, Claude P, Pierre P, Georgette L (1985) Antibacterial activities of erythromycins A, B, C, and D and some of their derivatives. Antimicrob Agents Chemother 28:630–633CrossRefGoogle Scholar
  2. 2.
    Chen Y, Deng W, Wu J, Qian J, Chu J, Zhuang Y, Zhang S, 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 74:1820–1828CrossRefGoogle Scholar
  3. 3.
    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 m(3) scale. Bioresour Technol 100:1406–1412CrossRefGoogle Scholar
  4. 4.
    Zhang Q, Chen Y, Hong M, Gao Y, Chu J, Zhuang YP, Zhang SL (2014) The dynamic regulation of nitrogen and phosphorus in the early phase of fermentation improves the erythromycin production by recombinant Saccharopolyspora erythraea strain. Bioresour Bioprocess 1:1–6CrossRefGoogle Scholar
  5. 5.
    Wang Y, 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 75:837–842CrossRefGoogle Scholar
  6. 6.
    Peter B, Wolfgang M, Pauli TK, James EB (1998) Genetic engineering of an industrial strain of Saccharopolyspora erythraea for stable expression of the Vitreoscilla haemoglobin gene (vhb). Microbiology 144:2441–2448CrossRefGoogle Scholar
  7. 7.
    Jean P, Paul P (1993) Influence of n-propanol on growth and antibiotic production by an industrial strain of Streptomyces erythreus under different nutritional condition. Biotechnol Lett 15:455–460CrossRefGoogle Scholar
  8. 8.
    Konstancja RB, Zbigniew R, Danuta SK, Andrzej R (1973) Limiting reaction in activation of acyl units in biosynthesis of macrolide antibiotic. Am Soc Microbiol 2:162–167Google Scholar
  9. 9.
    Murli S, Kennedy J, Dayem LC, Carney JR, Kealey JT (2003) Metabolic engineering of Escherichia coli for improved 6-deoxyerythronolide B production. J Ind Microbiol Biotechnol 30:500–509CrossRefGoogle Scholar
  10. 10.
    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 36:1445–1453CrossRefGoogle Scholar
  11. 11.
    Reeves AR, Cernota WH, Brikun IA, Wesley RK, Weber JM (2004) Engineering precursor flow for increased erythromycin production in Aeromicrobium erythreum. Metab Eng 6:300–312CrossRefGoogle Scholar
  12. 12.
    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 33:600–609CrossRefGoogle Scholar
  13. 13.
    Weber JM, Cernota WH, Gonzalez MC, Leach BI, Reeves AR, Wesley RK (2012) An erythromycin process improvement using the diethyl methylmalonate responsive (Dmr) phenotype of the Saccharopolyspora erythraea mutB strain. Appl Microbiol Biotechnol 93:1575–1583CrossRefGoogle Scholar
  14. 14.
    Wu JQ, Zhang QL, Deng W, Qian JC, Zhang SL, Liu W (2011) Toward improvement of erythromycin A production in an industrial Saccharopolyspora erythraea strain via facilitation of genetic manipulation with an artificial attB site for specific recombination. Appl Environ Microbiol 77:7508–7516CrossRefGoogle Scholar
  15. 15.
    Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49CrossRefGoogle Scholar
  16. 16.
    Hopwood DA, Bibb MJ, Bruton CJ, Kieser T, Chater KF, Smith CP, Kieser HM, Lydiate DJ, Ward JM, Schempf H (2000) Genetic manipulation of streptomyces: a laboratory manual. The John Innes Foundation, NorwichGoogle Scholar
  17. 17.
    Matsushima P, Broughton MC, Turner JR, Baltz RH (1994) Conjugal transfer of cosmid DNA from Escherichia coli to Saccharopolyspora spinosa: effects of chromosomal insertions on macrolide A83543 production. Gene 146:39–45CrossRefGoogle Scholar
  18. 18.
    Linde T, Zoglowek M, Lubeck M, Frisvad JC, Lubeck PS (2016) The global regulator LaeA controls production of citric acid and endoglucanases in Aspergillus carbonarius. J Ind Microbiol Biotechnol. doi: 10.1007/s10295-016-1781-3 Google Scholar
  19. 19.
    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 100:3358–3365CrossRefGoogle Scholar
  20. 20.
    Chen Y, Wang ZJ, Chu J, Xi BL, Zhuang YP (2015) The glucose RQ-feedback control leading to improved erythromycin production by a recombinant strain Saccharopolyspora erythraea ZL1004 and its scale-up to 372-m(3) fermenter. Bioprocess Biosyst Eng 38:105–112CrossRefGoogle Scholar
  21. 21.
    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 134:173–179CrossRefGoogle Scholar
  22. 22.
    Liu TT, Wang T, Yang Y, Wang ZJ, Zhuang YP, Chu J, Guo MJ (2016) Low field nuclear magnetic resonance for rapid quantitation of microalgae lipid and its application in high throughput screening. Chin J Biotech. doi: 10.13345/j.cjb.150489 Google Scholar
  23. 23.
    Parekh S, Vinci VA, Strobel RJ (2000) Improvement of microbial strains and fermentation processes. Appl Microbiol Biotechnol 54:287–301CrossRefGoogle Scholar
  24. 24.
    Baltz RH (2006) Molecular engineering approaches to peptide, polyketide and other antibiotics. Nat Biotechnol 24:1533–1540CrossRefGoogle Scholar
  25. 25.
    Tsuji K, Goetz JF (1978) HPLC as a rapid means of monitoring erythromycin and tetracycline fermentation processes. J Antibiot 31:302–308CrossRefGoogle Scholar
  26. 26.
    Licona-Cassani C, Marcellin E, Quek LE, Jacob S, Nielsen LK (2012) Reconstruction of the Saccharopolyspora erythraea genome-scale model and its use for enhancing erythromycin production. Antonie Van Leeuwenhoek 102:493–502CrossRefGoogle Scholar
  27. 27.
    Zhang G, Yang G, Wang X, Guo Q, Li Y, Li J (2012) Influence of blocking of 2,3-butanediol pathway on glycerol metabolism for 1,3-propanediol production by Klebsiella oxytoca. Appl Biochem Biotechnol 168:116–128CrossRefGoogle Scholar
  28. 28.
    Seo MY, Seo JW, Heo SY, Baek JO, Rairakhwada D, Oh BR, Seo PS, Choi MH, Kim CH (2009) Elimination of by-product formation during production of 1,3-propanediol in Klebsiella pneumoniae by inactivation of glycerol oxidative pathway. Appl Microbiol Biotechnol 84:527–534CrossRefGoogle Scholar
  29. 29.
    Nocon J, Steiger M, Mairinger T, Hohlweg J, Russmayer H, Hann S, Gasser B, Mattanovich D (2016) Increasing pentose phosphate pathway flux enhances recombinant protein production in Pichia pastoris. Appl Microbiol Biotechnol. doi: 10.1007/s00253-016-7363-5 Google Scholar
  30. 30.
    Siedler S, Lindner SN, Bringer S, Wendisch VF, Bott M (2013) Reductive whole-cell biotransformation with Corynebacterium glutamicum: improvement of NADPH generation from glucose by a cyclized pentose phosphate pathway using pfkA and gapA deletion mutants. Appl Microbiol Biotechnol 97:143–152CrossRefGoogle Scholar
  31. 31.
    Tao YZ, Liu D, Yan X, Zhou ZH, Lee JK, Yang C (2012) Network identification and flux quantification of glucose metabolism in Rhodobacter sphaeroides under photoheterotrophic H-2-producing conditions. J Bacteriol 194:274–283CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiPeople’s Republic of China

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