Repository hosted by TU Delft Library

Home · Contact · About · Disclaimer ·
 

An expression system based on the promoter region of the Aspergillus awamori 1,4-bèta-endoxylanase A gene

Publication files not online:

Author: Gouka, R.J. · Hessing, J.G.M. · Punt, P.J. · Stam, H. · Musters, W. · Hondel, C.A.M.J.J. van den
Type:article
Date:1996
Institution: TNO Voeding
Source:Appied Microbiology and Biotechnology, 46, 28-35
Identifier: 68889
doi: doi:10.1007/s002530050779
Keywords: Nutrition · Enzyme · Glycosidase · Microbial enzyme · Controlled study · Gene expression regulation · Molecular cloning · Nonhuman · Promoter region · Aspergillus · Endo-1,4-beta Xylanases · Enzyme Induction · Gene Expression Regulation, Fungal · Genes, Reporter · Genetic Vectors · Glucan 1,4-alpha-Glucosidase · Glucuronidase · Promoter Regions (Genetics) · RNA, Fungal · RNA, Messenger · Transcription, Genetic · Xylose · Xylosidases · Aspergillus awamori · Aspergillus niger · Trixis

Abstract

A new, highly inducible fungal promoter derived from the Aspergillus awamori 1,4-β-endoxylanase A (exlA) gene is described. Induction analysis, carried out with the wild-type strain in shake flasks, showed that exlA expression is regulated at the transcriptional level. Using a β-glucuronidase (uidA) reporter strategy, D-xylose was shown to be an efficient inducer of the exlA promoter, whereas sucrose or maltodextrin were not. Upon D-xylose induction, the exlA promoter was threefold more efficient than the frequently used A. niger glucoamylase (glaA) promoter under maltodextrin induction. Detailed induction analyses demonstrated that induction was-dependent on the presence of D-xylose in the medium. Carbon-source-limited chemostat cultures with the uidA reporter strain showed that D-xylose was also a very good inducer in a fermenter, even in the presence of sucrose.