Purification to homogeneity of Candida albicans glucosamine-6-phosphate synthase overexpressed in Escherichia coli.
The Candida albicans GFA1 gene encoding glucosamine-6-phosphate synthase, an enzyme of cell wall biosynthesis pathway in fungi and bacteria, recently an object of interest as a target for the chemotherapy of systemic mycoses, was PCR amplified and cloned to an Escherichia coli expression vector pET23b. The activity of the enzyme in the lysates from the overproducing E. coli strain was approximately 50-100 times higher than in the lysates from the control E. coli strain. This abundant overproduction allows to purify milligram amounts of the enzyme to homogeneity.
Department of Microbiology, Department of Pharmaceutical Technology and Biochemistry, Technical University of Gdańsk, ul. G. Narutowicza 11/12, Gdańsk, 80-952, Poland. psach@altis.chem.pg.gda.pl
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Sachadyn
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Jedrzejczak
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Milewski
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Kur
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Borowski
D002176:Candida albicans / Q000201:enzymology / Q000235:genetics*; D011232:Chemical Precipitation; D002845:Chromatography; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005800:Genes, Fungal; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing) / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D004271:DNA, Fungal; D011994:Recombinant Proteins; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)