NMMP

Minimal liquid medium (NMMP)

NMMP medium is used to obtain dispersed growth of Streptomyces sp. with specific carbon sources.^[2]

Preparation

Make up the following solutions:

Minor elements solution: (per litre)

• 1g ZnSO₄*7H₂O
• 1g FeSO₄*7H₂O
• 1g MnCl₂*4H₂O
• 1g CaCl₂, anhydrous

Phosphate buffer

• Mix 0.1M solutions of NaH₂PO₄ and K₂HPO₄ to give pH 6.8

Carbon source

• 20% solution, sterilized separately

NMMP

• 2 g (NH₄)₂SO₄
• 5 g Difco Casaminoacids
• 0.6g MgSO₄*7H₂O
• 50g PEG 6000
• 1 mL minor elements solution
• 800 mL ddH₂O

Instructions

1. Make up the NMMP solution described above
2. Dispense in 80 mL aliquots
3. Autoclave
4. Make up the phosphate buffer solution, carbon source, and any other required growth factors, and autoclave these separately

Just before using, add:

• 15 mL NaH₂PO₄/K₂HPO₄ buffer (0.1M, pH 6.8)
• 2.5 mL carbon source (20%)
• 2.5 mL any other required growth factors (add 2.5 mL sterile water if none)

Uses

• Routine growth of Streptomyces sp. in liquid culture

Notes

• Using PEG-6000 and adding stainless steel springs in the culture flasks allows for good dispersed growth of many Streptomyces strains.^[1]

• Agar can be added (20 g/L) to make solid NMMP.^[3]

• A modified version of NMMP suitable for ¹⁴N/¹⁵N labelling has been developed.^[4]

References

[1] Hodgson (1982). Glucose Repression of Carbon Source Uptake and Metabolism in Streptomyces coelicolor A3(2) and its Perturbation in Mutants Resistant to 2-Deoxyglucose. Journal of General Microbiology 128: 2417-2430.

[2] Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK

[3] Rico, S., Yepes, A., Rodríguez, H., Santamaría, J., Antoraz, S., Krause, E. M., … Santamaría, R. I. (2014). Regulation of the AbrA1/A2 two-component system in Streptomyces coelicolor and the potential of its deletion strain as a heterologous host for antibiotic production. PloS one, 9(10), e109844. doi:10.1371/journal.pone.0109844

[4] Świątek, M. A., Gubbens, J., Bucca, G., Song, E., Yang, Y. H., Laing, E., … van Wezel, G. P. (2013). The ROK family regulator Rok7B7 pleiotropically affects xylose utilization, carbon catabolite repression, and antibiotic production in Streptomyces coelicolor. Journal of bacteriology, 195(6), 1236–1248. doi:10.1128/JB.02191-12