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High-level expression of Humicola insolens cutinase in Pichia pastoris without carbon starvation and its use in cotton fabric bioscouring
- Hong, Ruoyu, Sun, Yirong, Su, Lingqia, Gu, Lengtao, Wang, Fuqing, Wu, Jing
- Journal of biotechnology 2019 v.304 pp. 10-15
- Humicola insolens, Komagataella pastoris, Thermobifida fusca, bioreactors, bioscouring, butyrates, carbon, cell growth, cotton fabric, cutinase, enzyme activity, fermentation, glycerol, methanol, monitoring, sorbitol, starvation, wettability
- Huimcola insolens cutinase (HiC) was heterologously expressed in Pichia pastoris. To avoid a carbon starvation step, fermentation was conducted using combinations of sorbitol with glycerol and methanol in the cell growth and induction phases, respectively. The cutinase productivity (27.71 U mL−1 h−1) was 9.93 U mL−1 h−1 greater than that achieved using traditional two-phase methods, and a cutinase activity of 2660 U mL−1, using p-nitrophenyl butyrate as substrate, was achieved after only 96 h in a 3-L bioreactor. Subsequently, the combination of HiC with Thermobifida fusca cutinase (TfC) in cotton fabric bioscouring was evaluated by monitoring the wettability and dyeability of the fabric. Treatment with 20 U mL−1 of HiC at 80 °C for 5 min followed by 30 U mL−1 of TfC at 50 °C for 1 h gave the best results. The total treatment time was shorter and performance was better than those seen with the alkali method.