An expression system based on the promoter region of the Aspergillus awamori 1,4-beta-endoxylanase A gene.
A new, highly inducible fungal promoter derived from the Aspergillus awamori 1,4-beta-endoxylanase A (exlA) gene is described. Induction analysis, carried out with the wild-type strain in shake flasks, showed that exlA expression in regulated at the transcriptional level. Using a beta-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.
Gouka
R J
RJ
TNO Nutrition and Food Research Institute, Department of Molecular Genetics and Gene Technology, Rijswijk, The Netherlands.
Hessing
J G
JG
Punt
P J
PJ
Stam
H
H
Musters
W
W
Van den Hondel
C A
CA
D001230:Aspergillus / Q000201:enzymology / Q000235:genetics*; D043364:Endo-1,4-beta Xylanases; D004790:Enzyme Induction / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic; D014994:Xylose / Q000494:pharmacology; D014995:Xylosidases / Q000235:genetics* / Q000378:metabolism
D012331:RNA, Fungal; D012333:RNA, Messenger; D014994:Xylose; D014995:Xylosidases; D005087:Glucan 1,4-alpha-Glucosidase; D005966:Glucuronidase; D043364:Endo-1,4-beta Xylanases