Additional file 1 of Production of transglutaminase in glutathione-producing recombinant Saccharomyces cerevisiae

Additional file 1: Figure S1. Cross-linking assay using BSA as a substrate after incubation for 120 min with several volumes of SmTG solution extracted from the BY4741-TG strain. Lane 1, molecular size marker; lane 2, BSA only; lanes 3, 4 and 5, 1×, 5× and 10×SmTG (+BSA), respectively; lanes 6, 7 and 8, 1×, 5× and 10×SmTG (−BSA), respectively. Figure S2. Effect of adding reduced glutathione to polymerization of caseinate by SmTG. SDS-PAGE profiles show the protein pattern of caseinate after incubation with SmTG. 1×SmTG extracted from the BY4741-TG strain was used. Lane 1, molecular size marker; lane 2, caseinate only; lanes 3, 4, 5, 6, 7 and 8, caseinate, SmTG and 0, 0.2, 0.4, 0.8, 1.6 and 3.2 mM glutathione, respectively; lane 9, SmTG only. The polymerized caseins (molecular weight > 100 kDa) presented in the top panel were recovered and measured as shown in Fig. 2. Figure S3. Growth and glutathione production of S. cerevisiae expressing SmTG. (a) Time course of cell growth. Solid and dotted lines represent the BY4741-TG and GCI-TG strains, respectively. (b) Glutathione concentrations in 1×SmTG solutions extracted from the BY4741-TG strain (black) and GCI-TG strain (white). (c) Expression level of SmTG after cultivation for 24 h. lane 1, BY4741-TG strain; lane 2, GCI-TG strain. Expression level of SmTG was calculated from the intensity of each SmTG band and represented as relative values (%). The values are means and the error bars show the SD (n = 3).