Autolysis of Pichia pastoris induced by cold

AMB Express

Cipriano Chávez-Cabrera¹  María E. Hidalgo-Lara¹  Yaneth Bartolo-Aguilar¹  Luis B. Flores-Cotera¹  Luc Dendooven¹  Rodolfo Marsch¹  Lourdes Villa-Tanaca² 

[1] Department of Biotechnology and Bioengineering, Cinvestav-IPN;Department of Microbiology, Escuela Nacional de Ciencias Biológicas del IPN;

关键词: Autolysis;    Cold-shock promoter;    Glucanase;    Saccharomyces cerevisiae;    Pichia pastoris;    White biotechnology;   

DOI  :  10.1186/s13568-017-0397-y

来源: DOAJ

【 摘 要 】

Abstract The production of recombinant biopharmaceutical proteins is a multi-billion dollar market. Protein recovery represents a major part of the production costs. Pichia pastoris is one of the microbial systems most used for the production of heterologous proteins. The use of a cold-induced promoter to express lytic enzymes in the yeast after the growth stage could reduce protein recovery costs. This study shows that a cold-shock can be applied to induce lysis of the yeast cells. A strain of P. pastoris was constructed in which the endogenous eng gene encoding a putative endo-β-1,3-glucanase was overexpressed using the cold-shock induced promoter of the cctα gene from Saccharomyces cerevisiae. In the transgenic P. pastoris, the expression of eng increased 3.6-fold after chilling the cells from 30 to 4 °C (cold-shock stage) followed by incubation for 6 h (eng expression stage). The culture was heated to 30 °C for 6 h (ENG synthesis stage) and kept at 37 °C for 24 h (lysis stage). After this procedure the cell morphology changed, spheroplasts were obtained and cellular lysis was observed. Thus, a clone of P. pastoris was obtained, which undergoes autolysis after a cold-shock.

【 授权许可】

Unknown