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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.