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Applied Microbiology and Biotechnology

, Volume 33, Issue 4, pp 395–400 | Cite as

Improved production of heterologous protein from Streptomyces lividans

  • Gregory F. Payne
  • Neslihan DelaCruz
  • Steven J. Coppella
Biotechnology

Summary

Protein-secreting procaryotic host organisms are currently being sought as alternatives to Escherichia coli for recombinant processing. In this study we examined how manipulation of the cultivation conditions can enhance heterologous protein production by Streptomyces lividans. The recombinant S. lividans used in this study expressed and excreted a Flavobacterium enzyme capable of hydrolyzing organophosphates. Initial shake-flask studies demonstrated that supplementing Luria-Bertani medium with moderate amounts of glucose (30 g/l), led to improved enzyme production. In fermentor studies with controlled pH, a further twofold increase in production was observed when glucose was fed continuously as compared to batch cultivation. This improved production in the glucose-fed culture may be related to a reduced accumulation of acids. Continuous feeding of both glucose and tryptone led to a further sixfold increase in production. In addition to enhancing production 25-fold, the efficiency of enzyme production and the specific activity of the excreted enzyme were also improved by glucose and tryptone feeding. These results demonstrate that in addition to genetic manipulations, optimization of cultivation conditions can lead to significant improvements in the production of heterologous proteins from Streptomyces.

Keywords

Glucose Streptomyces Enzyme Production Batch Cultivation Genetic Manipulation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Ahmed ZU, Shapiro S, Vining LC (1984) Excretion of α-keto acids by strains of Streptomyces venezuelae. Can J Microbiol 30:1014–1021Google Scholar
  2. Bertrand J-L, Morosili R, Shareck F, Kluepfel D (1989) Expression of the xylanase gene of Streptomyces lividans and production of the enzyme on natural substrates. Biotechnol Bioeng 33:791–794Google Scholar
  3. Brown KA (1980) Phosphotriesterases of Flavobacterium sp. Soil Biol Biochem 12:105–112Google Scholar
  4. Crawford DL (1988) Development of recombinant Streptomyces for biotechnological and environmental uses. Biotechnol Adv 6:183–206Google Scholar
  5. Dekleva ML, Strohl WR (1987) Glucose-stimulated acidogenesis in Streptomyces peucetius. Can J Microbiol 33:1129–1132Google Scholar
  6. Doskocil J, Hostalek Z, Kasparova J, Zajicek J, Herold M (1959) Development of Streptomyces aureofaciens in submerged culture. J Biochem Microbiol Technol Eng 1:261–271Google Scholar
  7. Ghangas GS, Wilson DB (1987) Expression of a Thermomonospora fusca cellulase gene in Streptomyces lividans and Bacillus subtilis. Appl Environ Microbiol 53:1470–1475Google Scholar
  8. Illingsworth C, Larson G, Hellekant G (1989) Secretion of sweet-tasting plant protein thaumatin by Streptomyces lividans. J Ind Microbiol 4:37–42Google Scholar
  9. Katz E, Thompson CJ, Hopwood DA (1983) Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol 129:2703–2714Google Scholar
  10. Mulbry WW, Karns JS (1989) Purification and characterization of three parathion hydrolasesfrom Gram-negative bacterial strains. Appl Environ Microbiol 55:289–293Google Scholar
  11. Munnecke DM, Fischer HF (1979) Production of parathion hydrolase activity. Eur J Appl Microbiol 8:103–112Google Scholar
  12. Serdar CM, Gibson DT (1985) Enzymatic hydrolysis of organophosphates: cloning and expression of a parathion hydrolase gene from Pseudomonas diminuta. Biotechnology 3:567–571Google Scholar
  13. Sethunathan N, Yoshida T (1973) A Flavobacterium sp. that degrades diazinon and parathion. Can J Microbiol 19:873–875Google Scholar
  14. Steiert JS, Pogell BM, Speedie MK, Laredo J (1989) A gene coding for a membrane-bound hydrolase is expressed as a secreted, soluble enzyme in Streptomyces lividans. Biotechnology 7:65–68Google Scholar
  15. Surowitz KG, Pfister RM (1985) Glucose metabolism and pyruvate excretion by Streptomyces alboniger. Can J Microbiol 31:702–706Google Scholar
  16. Wang DIC (1988) Biotechnology: status and perspectives. American Institute of Chemical Engineers Monograph Series, vol 84. American Institute of Chemical Engineers, New York, pp 4–10Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Gregory F. Payne
    • 1
    • 2
  • Neslihan DelaCruz
    • 1
  • Steven J. Coppella
    • 1
    • 3
  1. 1.Department of Chemical and Biochemical EngineeringUniversity of MarylandBaltimoreUSA
  2. 2.Center for Agricultural BiotechnologyUniversity of MarylandBaltimoreUSA
  3. 3.Medical Biotechnology CenterUniversity of MarylandBaltimoreUSA

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