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Toxic misfolding of Arabidopsis cellulases in the secretory pathway of Pichia pastoris.

Abstract：

:Plants produce a large number of cellulases that are either secreted or anchored in the plasma membrane where they likely function in various aspects of cellulose synthesis, modification and degradation during plant growth and development. Very few of these enzymes have been characterized in any detail, however. Here we attempted to produce two Arabidopsis modular cellulases, which contain a catalytic domain belonging to glycoside hydrolase family 9 (GH9) and a carbohydrate binding module (CBM), in the yeast Pichia pastoris. Neither of the intact modular enzymes was detectably produced, although the independently expressed GH9 catalytic domain of one enzyme was secreted when the protein was expressed at low temperature. Expression of intact and truncated cellulases at the standard temperature caused extensive cell lysis, with release of high concentrations of endogenous proteins into the culture medium. Cell lysis appeared to result from misfolding of cellulase proteins within the Pichia secretory pathway. The toxicity of these misfolded cellulases potentially could be exploited to derive host strains with enhanced capability to fold recombinant secretory proteins.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Tawde MD,Freimuth P

doi

10.1016/j.pep.2012.08.009

subject

Has Abstract

pub_date

2012-10-01 00:00:00

pages

211-7

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(12)00218-5

journal_volume

85

pub_type

杂志文章

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