Chlorella pyrenoidosa (C. pyrenoidosa) contains Chlorella Growth Factor (CGF), which consists of proteins and polysaccharides. CGF is located inside the nucleus of cells and is beneficial to humans as a food supplement, an immunity booster, and an antioxidant. CGF formation of C. pyrenoidosa is influenced by medium composition. C. pyrenoidosa INK was cultured in a modified basal medium (MBM) with various concentrations of Mg2+ (0.5, 1.0, and 1.5 g/L) and Fe2+ (3.5×10-4 and 5.0×10-4 g/L). The experiments were performed and analyzed under a completely randomized design using a 2-L bottle with three replications. The results showed that MBM with 1.0 g/L of Mg2+ and 3.5×10-4 g/L of Fe2+ yielded the optimal growth curve for C. pyrenoidosa. Analysis of protein content was carried out using the Lowry method with a spectrophotometer at λ=750 nm, and the obtained results were 0.0974 mg/mL (extract) and 6.4097 mg/ml (supernatant). Furthermore, analysis of glucose content was carried out using the phenol sulfate method (λ = 490 nm), and the obtained results were 49.331 ppm (extract) and 1566.911 ppm (supernatant). Analysis of amino acids in CGF using liquid chromatography-mass spectrometry (LC-MS) indicated the presence of tyrosine, proline, glutamate, alanine, valine, tryptopan, phenylalanine, methionine, and leucine-isoleucine.



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