Fig. 2.

Generation of SEAP-producing P. pastoris and SEAP production in hydrogel system. (A) Comparison of SEAP production between 48 h and 72 h fermentation at 30 °C. P. pastoris Pp01 and Pp02 were used as the control and SEAP-producing strain, respectively. 0.5% methanol was added every 24 h to maintain induction (n = 3 for each strain). (B) SDS-PAGE analysis of recombinant SEAP production. The protein marker was loaded into M lane of SDS-PAGE and supernatant of pelleted yeast containing SEAP was loaded into S lane. The red arrow indicates the size of SEAP (around 60 kDa) secreted to the media. (C) SEAP production assessed both pre- and post-lyophilization (data from round 2 and round 5 of reuse respectively) from hydrogels. (D) SEAP production in hydrogels (blue bars) with repeated use compared to liquid culture (orange bars) performance. All reuse batches were carried out with 30 °C incubation for 48 h. All the samples were treated with lyophilization after round 3 of reuse. Data are mean ± s.d.; n = 6 biological replicates for hydrogels n = 3 biological replicates for liquid culture. *P < .05; **P < .01; ***P < .001 via two-sample t-test (hydrogel vs corresponding liquid culture for each round of reuse).