3.5 Validation by supplemental cultivation
We performed validation cultivations to confirm the estimation of important components by DNN-MIE. Figure 5 demonstrates the results of these validation experiments. Glu, maltose, Alanine (Ala), phenylalanine (Phe), Ile, trehalose, Lys, Asp, phosphate, Gly, and sorbitol significantly increased GFP yields, and leucine (Leu), Serine (Ser), threonine (Thr), asparagine (Asn), valine (Val), glycerol, and tyrosine (Tyr) significantly decreased GFP yields. In particular, because Glu increased the GFP yields to 112.9 ± 4.0% and the cell yields to 104.8 ± 1.5%, this amino acid was predicted to be the most important variable. Maltose stimulated the GFP yields by 106.5 ± 1.2% and the cell yields by 104.9 ± 1.1%. Asp induced less than 2% of the cell and GFP yields. Sorbitol showed no influence on the cell and GFP yields. The final pH was between 6.44 and 6.54 in all cases.