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Non-classical secretion of 1,4-alpha-glucan branching enzymes without signal peptides in Escherichia coli.

Abstract：

:1,4-α-Glucan branching enzyme (GBE, EC. 2.4.1.18), which plays a key role in the synthesis of starch and glycogen, has been overexpressed in E. coli as an intracellular enzyme by many researchers. In this study, it was found that the GBEs from Geobacillus thermoglucosidans and Rhodothermus obamensis were secreted into the culture medium when they were expressed separately, in E. coli. This occurred despite the absence of any signal peptide. In fact, although bioinformatics tools predicted that both of these proteins would localize to the cytoplasm, a high level of expression and non-classical secretion was found to achieve without addition of the inducer isopropyl β-d-thiogalactopyranoside. Further experiments revealed that secretion was a two-step process that occurred via the periplasmic space. Results excluded the involvement of the Sec pathway or the TAT pathway. Instead, the findings indicated a relationship between cell membrane integrity and the secretion of the two GBEs, and suggested that their N-termini play an essential role in their expression and secretion.

journal_name

Int J Biol Macromol

journal_title

International journal of biological macromolecules

authors

Xin C,Ban X,Gu Z,Li C,Cheng L,Hong Y,Li Z

doi

10.1016/j.ijbiomac.2019.04.002

subject

Has Abstract

pub_date

2019-07-01 00:00:00

pages

759-765

eissn

0141-8130

issn

1879-0003

pii

S0141-8130(18)35281-4

journal_volume

132

pub_type

杂志文章

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