Title
Kinetics of mRNA and protein synthesis of genes controlled by the 1,4- β-endoxylanase A promoter in controlled fermentations of Aspergillus awamori
Author
Gouka, R.J.
Stam, H.
Fellinger, A.J.
Muijsenberg, R.J.G.T.
van de Wijngaard, A.J.
Punt, P.J.
Musters, W.
van den Hondel, C.A.M.J.J.
TNO Voeding
Publication year
1996
Abstract
In this study, induction and repression kinetics of the expression of the Aspergillus awamori 1,4-β-endoxylanase A (exlA) gene under defined physiological conditions was analyzed at the mRNA and the protein levels. Induction was analyzed by pulsing D-xylose to a sucrose-limited continuous culture of an A. awamori 1,4-β-endoxylanase A (EXLA)-overproducing strain. Directly after the D-xylose pulse, exlA mRNA was synthesized, and it reached a constant maximal level after 45 to 60 min. This level was maintained as long as D-xylose was present. The kinetics of mRNA synthesis of the genes encoding Thermomyces lanuginosa lipase (lplA) and Escherichia coli β- glucuronidase (uidA), which were also under the control of the exlA promoter, were similar to those observed for exlA mRNA. The repression of exlA expression was analyzed by pulsing D-glucose to a D-xylose-limited continuous culture. Immediately after the glucose pulse, the exlA mRNA level declined rapidly, with a half-life of approximately 20 to 30 min, and it reached a minimal level after 60 to 90 min. The time span between mRNA synthesis and the secretion of proteins was determined for EXLA and lipase. In both cases, mRNA became visible after approximately 7.5 min. After 1 h, both proteins became detectable in the medium but the rate of secretion of EXLA was faster than that of lipase.
Subject
Nutrition
Triacylglycerol lipase
Cell culture
Enzyme activity
Enzyme analysis
Gene expression
Gene expression regulation
Glucose metabolism
Promoter region
Protein synthesis
Rna analysis
Aspergillus
Endo-1,4-beta Xylanases
Fermentation
Gene Expression Regulation, Fungal
Glucose
Glucuronidase
Kinetics
Lipase
Promoter Regions (Genetics)
Protein Biosynthesis
Recombinant Fusion Proteins
RNA, Messenger
Xylose
Xylosidases
Aspergillus awamori
Escherichia coli
Thermomyces lanuginosus
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http://resolver.tudelft.nl/uuid:81bff37b-bf70-4453-8790-60618ce3182e
TNO identifier
233485
ISSN
0099-2240
Source
Applied and Environmental Microbiology, 62 (10), 3646-3649
Document type
article