The Arxula adeninivorans ATAL gene encoding transaldolase-gene characterization and biotechnological exploitation.
The yeast Arxula adeninivorans provides an attractive expression platform and can be exploited as gene source for biotechnologically interesting proteins. In the following study, a striking example for the combination of both aspects is presented. The transaldolase-encoding A. adeninivorans ATAL gene, including its promoter and terminator elements, was isolated and characterized. The gene includes a coding sequence of 963 bp encoding a putative 321 amino acid protein of 35.0 kDa. The enzyme characteristics analyzed from isolates of native strains and recombinant strains overexpressing the ATAL gene revealed a molecular mass of ca. 140 kDa corresponding to a tetrameric structure, a pH optimum of ca. 5.5, and a temperature optimum of 20 degrees C. The preferred substrates for the enzyme include D-erythrose-4-phosphate and D-fructose-6-phosphate, whereas D-glyceraldehyde is not converted. The ATAL expression level under salt-free conditions was observed to increase in media supplemented with 5% NaCl rendering the ATAL promoter attractive for moderate heterologous gene expression under high-salt conditions. Its suitability was assessed for the expression of a human serum albumin (HSA) reporter gene.
Fiki
Ayman El
AE
Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstr. 3, 06466 Gatersleben, Germany.
Metabteb
Gamal El
GE
Bellebna
Carmen
C
Wartmann
Thomas
T
Bode
Rüdiger
R
Gellissen
Gerd
G
Kunze
Gotthard
G
D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D004718:Saccharomycetales / Q000201:enzymology / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology; D014153:Transaldolase / Q000235:genetics* / Q000378:metabolism
D005656:Fungal Proteins; D011994:Recombinant Proteins; D012965:Sodium Chloride; D014153:Transaldolase