Recombinant B. choshinensis grew efficiently in Trichormus extracts using the shaking method, similar to growth in conventional medium (CM) (A: macroscopic observation; B: turbidity analysis). B. choshinensis consumed small amounts of glucose (C) and pyruvate (D) and actively consumed proteinogenic amino acids (E and F) in the Trichormus extract. It also secreted bovine beta-casein, human alpha-lactalbumin, human insulin-like growth factor-1 (IGF-1), and human albumin when cultivated in the Trichormus extract, similar to the CM (G). Alpha-lactoglobulin is represented by the concentration of the His-tag fusion protein. Data are presented as mean ± standard deviation (n= 3). For the statistical analysis of glucose, pyruvate, and total amino acid concentrations, comparisons were made between (1) the amounts with and without microorganism culture and (2) each amount without microorganism culture. See also Table S10. TE (S), Trichormus extracts by the shaking method, *p< 0.05, **p< 0.01, ***p< 0.001, ****p< 0.0001.
