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Autolysis of Pichia pastoris induced by cold.

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.

journal_name

AMB Express

journal_title

AMB Express

authors

Bartolo-Aguilar Y,Dendooven L,Chávez-Cabrera C,Flores-Cotera LB,Hidalgo-Lara ME,Villa-Tanaca L,Marsch R

doi

10.1186/s13568-017-0397-y

subject

Has Abstract

pub_date

2017-12-01 00:00:00

pages

95

issue

1

issn

2191-0855

pii

10.1186/s13568-017-0397-y

journal_volume

7

pub_type

杂志文章

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