The cloning and expression of a gene encoding old yellow enzyme from Saccharomyces carlsbergensis

Kazuyo Saito, Dennis J. Thiele, Maryann Davio, Oksana Lockridge, Vincent Massey

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

We have identified a gene that encodes Old Yellow Enzyme in brewer's bottom yeast. The open reading frame encodes a polypeptide of 400 amino acids with Mr = 45,021. Using the T7 RNA polymerase system, recombinant enzyme was expressed in Escherichia coli. 17 mg of Old Yellow Enzyme was obtained from a 3-liter cell culture, and the recombinant enzyme had NADPH oxidase activity. On fast protein liquid chromatography separation, the recombinant enzyme showed a single large peak, while native enzyme from brewer's bottom yeast separated into five fractions on fast protein liquid chromatography. Southern blot analysis showed that there are at least two Old Yellow Enzyme genes in brewer's bottom yeast genomic DNA. These results suggest that the heterogeneity of Old Yellow Enzyme in Saccharomyces carlsbergensis is due to the presence of multiple genes.

Original languageEnglish (US)
Pages (from-to)20720-20724
Number of pages5
JournalJournal of Biological Chemistry
Volume266
Issue number31
StatePublished - Nov 5 1991

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NADPH Dehydrogenase
Saccharomyces
Gene encoding
Cloning
Organism Cloning
Yeast
Saccharomyces cerevisiae
Gene Expression
Genes
Liquid chromatography
Enzymes
Liquid Chromatography
NADPH Oxidase
Southern Blotting
Cell culture
Escherichia coli
Open Reading Frames
Proteins
Cell Culture Techniques
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

The cloning and expression of a gene encoding old yellow enzyme from Saccharomyces carlsbergensis. / Saito, Kazuyo; Thiele, Dennis J.; Davio, Maryann; Lockridge, Oksana; Massey, Vincent.

In: Journal of Biological Chemistry, Vol. 266, No. 31, 05.11.1991, p. 20720-20724.

Research output: Contribution to journalArticle

Saito, Kazuyo ; Thiele, Dennis J. ; Davio, Maryann ; Lockridge, Oksana ; Massey, Vincent. / The cloning and expression of a gene encoding old yellow enzyme from Saccharomyces carlsbergensis. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 31. pp. 20720-20724.
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