Browse by author
Lookup NU author(s): Professor Jeremy Lakey,
Professor Colin Harwood
Full text for this publication is not currently held within this repository. Alternative links are provided below where available.
Following their secretion across the cytoplasmic membrane, processed secretory proteins of Bacillus subtilis must fold into their native conformation prior to translocation through the cell wall and release into the culture medium. The rate and efficiency of folding are critical in determining the yields of intact secretory proteins. The B. subtilis membrane is surrounded by a thick cell wall comprising a heteropolymeric matrix of peptidoglycan and anionic polymers. The latter confer a high density of negative charge on the wall, endowing it with ion-exchange properties, and secretory proteins destined for the culture medium must traverse the wall as the last stage in the export process. To determine the influence of charge on late stages in the secretion of proteins from this bacterium, we have used sequence data from two related α-amylases, to engineer the net charge of AmyL, an α-amylase from Bacillus licheniformis that is normally secreted efficiently from B. subtilis. While AmyL has a pI of 7.0, chimaeric enzymes with pI values of 5.0 and 10.0 were produced and characterized. Despite the engineered changes to their physico-chemical properties, the chimaeric enzymes retained many of the enzymic characteristics of AmyL. We show that the positively charged protein interacts with the cell wall in a manner that influences its secretion.
Author(s): Harwood CR; Lakey JH; Stephenson K; Jensen CL; Jorgensen ST
Publication type: Article
Publication status: Published
Journal: Biochemical Journal
ISSN (print): 0264-6021
ISSN (electronic): 1470-8728
Publisher: Portland Press
PubMed id: 10926823
Altmetrics provided by Altmetric