Applied Microbiology and Biotechnology

, Volume 78, Issue 5, pp 827–832 | Cite as

Defined salt formulations for the growth of Salinispora tropica strain NPS21184 and the production of salinosporamide A (NPI-0052) and related analogs

  • Ginger Tsueng
  • Sy Teisan
  • Kin S. Lam
Applied Microbial and Cell Physiology


Salinosporamide A (NPI-0052) is currently produced by a marine actinomycete, Salinispora tropica, via a saline fermentation process using a non-defined, commercially available synthetic sea salt, Instant Ocean. In order to control the consistency of the production of NPI-0052 and related analogs, two chemically defined salt formulations were developed to replace Instant Ocean. A chemically defined sodium-chloride-based salt formulation with similar sodium and chloride contents as in Instant Ocean was found to support higher production of NPI-0052 and a better metabolite production profile for downstream processing than Instant Ocean. A chemically defined sodium-sulfate-based salt formulation with low chloride concentration at 17 mM was found to support a similar NPI-0052 and metabolite production profile as Instant Ocean. The sodium-sulfate-based formulation is a robust formulation for large-scale production process due to its reduced corrosiveness in fermentation as compared with the saline fermentation utilizing Instant Ocean or the sodium-chloride-based salt formulation. The production of NPI-0052 in both chemically defined salt formulations was successfully scaled-up to a 42-l fermentor, indicating that these salt formulations can be used for large-scale manufacturing process.


Salinosporamide A NPI-0052 Salinispora tropica Saline fermentation Salt formulation 


  1. Barclay WR (2002) Reducing corrosion in a fermentor by providing sodium with a non-chloride sodium salt. US Patent 6,410,281Google Scholar
  2. Bidwell JP, Spotte S (1985) Artificial seawaters, formulas and methods. Jones and Bartlett, BostonGoogle Scholar
  3. Boulyga SF, Klötzli U, Stingeder G, Prohaska T (2007) Optimization and application of ICPMS with dynamic reaction cell for precise determination of 44Ca/40Ca isotope ratios. Anal Chem 79:7753–7760CrossRefGoogle Scholar
  4. Chauhan D, Catley L, Li G, Podar K, Hideshima T, Velankar M, Mitsiades C, Mitsiades N, Yasui H, Letai A, Ovaa H, Berkers C, Nicholson B, Chao TH, Neuteboom STC, Richardson P, Palladino MA, Anderson KC (2005) A novel orally active proteasome inhibitor induces apoptosis in multiple myeloma cells with mechanisms distinct from Bortezomib. Cancer Cell 8:407–419CrossRefGoogle Scholar
  5. Chauhan D, Hideshima T, Anderson KC (2006) A novel proteasome inhibitor NPI-0052 as an anticancer therapy. Brit J Cancer 95:961–965CrossRefGoogle Scholar
  6. Cruz RPV, Nishikata A, Tsuru T (1998) Pitting corrosion mechanism of stainless steels under wet–dry exposure in chloride-containing environments. Corrosion Sci 40:125–139CrossRefGoogle Scholar
  7. Cubadda F, Raggi A, Coni E (2006) Element fingerprinting of marine organisms by dynamic reaction cell inductively coupled plasma mass spectrometry. Anal Bioanal Chem 384:887–896CrossRefGoogle Scholar
  8. Cusack JC, Liu R, Xia L, Chao T-H, Pien C, Niu W, Palomebella VJ, Neuteboom STC, Palladino MA (2006) NPI-0052 enhances tumoricidal response to conventional cancer therapy in a colon cancer model. Clin Cancer Res 12:6758–6764CrossRefGoogle Scholar
  9. Feling RH, Bauchanan GO, Mincer TJ, Kaufman CA, Jensen PR, Fenical W (2003) Salinosporamide A: a highly cytotoxic proteasome inhibitor from a novel microbial source, a marine bacterium of the new genus Salinispora. Chem Int Ed 42:355–357CrossRefGoogle Scholar
  10. Fenical W, Jensen PR (2006) Developing a new resource for drug discovery: marine actinomycete bacteria. Nat Chem Biol 2:666–673CrossRefGoogle Scholar
  11. Groll M, Huber R, Potts BCM (2006) Crystal structures of salinosporamide A (NPI-0052) and B (NPI-0047) in complex with the 20S proteasome reveal important consequences of beta-lactone opening and a mechanism for irreversible binding. J Am Chem Soc 128:5136–5141CrossRefGoogle Scholar
  12. Jensen PR, Dwight R, Fenical W (1991) Distribution of actinomycetes in near-shore tropical marine sediments. Appl Environ Microbiol 57:1102–1108Google Scholar
  13. Macherla VR, Mitchell SS, Manam RR, Reed KA, Chao T, Nicholson B, Deyanat-Yazdi G, Mai B, Jensen PR, Fenical W, Neuteboom STC, Lam KS, Palladino MA, Potts BCM (2005) Structure–activity relationship studies of NPI-0052, a novel marine derived 20S proteasome inhibitor. J Med Chem 48:3684–3687CrossRefGoogle Scholar
  14. Maldonado LA, Fenical W, Jensen PR, Kauffman CA, Mincer TJ, Ward AC, Bull AT, Goodfellow M (2005) Salinispora arenicola gen. nov., sp. nov. and Salinispora tropica sp. nov., obligate marine actinomycetes belonging to the family Micromonosporaceae. Int J Syst Evol Microbiol 55:1759–1766CrossRefGoogle Scholar
  15. Mincer TJ, Jensen PR, Fenical W (2002) Widespread and persistent populations of a major new marine actinomycete taxon in ocean sediments. Appl Environ Microbiol 61:3695–3700Google Scholar
  16. Pilson MEQ (1998) An introduction to the chemistry of the sea. Prentice Hall, New JerseyGoogle Scholar
  17. Reed KA, Manam RR, Mitchell SS, Xu J, Teisan S, Chao TH, Deyanat-Yazdi G, Neuteboom STC, Lam KS, Potts BCM (2007) New salinosporamides from the marine actinomycete Salinispora tropica and thioester derivatives, potent inhibitors of 20S proteasome. J Nat Prod 70:269–276CrossRefGoogle Scholar
  18. Schweitzer PA (1995) Corrosion resistance tables, 4th edn. Marcel Dekker, New YorkGoogle Scholar
  19. Sedriks JA (1996) Corrosion of stainless steels, 2nd edn. Wiley, New YorkGoogle Scholar
  20. Tsaur C-C, Rock JC, Wang C-J, Su T-H (2005) The hot corrosion of 310 stainless steel with pre-coated NaCl/Na2SO4 mixtures at 750°C. Mat Chem Phys 89:445–453CrossRefGoogle Scholar
  21. Tsueng G, Lam KS (2007) Stabilization effect of resin on the production of potent proteasome inhibitor NPI-0052 during submerged fermentation of Salinispora tropica. J Antibiot 60:469–472CrossRefGoogle Scholar
  22. Tsueng G, McArthur KA, Potts BCM, Lam KS (2007) Unique butyric acid incorporation patterns for salinosporamides A and B reveal distinct biosynthetic origins. Appl Microbiol Biotechnol 75:999–1005CrossRefGoogle Scholar
  23. Udwary DW, Zeigler L, Asolkar RN, Singan V, Lapidus A, Fenical W, Jensen PR, Moore BS (2007) Genome sequence reveals complex secondary metabolome in the marine actinomycete Salinispora tropica. Proc Natl Acad Sci U S A 104:10376–10381CrossRefGoogle Scholar
  24. Vandamme EJ (1985) Biotechnology of industrial antibiotics. Marcel Dekker, New YorkGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Nereus Pharmaceuticals, Inc.San DiegoUSA

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