The use of culture media is necessary in growing microorganisms for the diagnosis of diseases, in research, and in many microbiological laboratory work. The study investigated the potential of Latundan banana Musa sapientum as sole nutrient source for microbial culture media. Banana powder was prepared from ripe and unripe banana fruit, and subjected to biochemical and chemical tests. Results revealed high amounts of carbohydrates, minerals (potassium, nitrogen, phosphorus, magnesium, calcium, and iron), and crude protein – all required for microbial growth. Solid media with different amounts of the banana powder were then formulated, and their capacity to sustain microbial growth for 24, 48 and 72 hours was tested using nutrient agar as positive control, and pure cultures of the bacteria Staphylococcus aureus and Pseudomonas aeruginosa and the fungus Aspergillus niger as test organisms. All banana media consistently sustained microbial growth, with the unripe fruit (16g/L) exhibiting the best growth in 72-hr period, which was comparable (no significant difference, p>0.01) to the growth of the same microorganisms on nutrient agar as positive control. Hence, unripe Latundan banana fruit may potentially be formulated as powder and commercialised as an alternative to nutrient agar in preparing microbial culture media.