Expression of a cDNA encoding human 5-lipoxygenase under control of the STA1 promoter in Saccharomyces cerevisiae

Gene
M NakamuraM Noma

Abstract

A yeast-expression vector utilizing the STA1 promoter was constructed, and shown to be useful for the expression of a heterologous gene. A cDNA, encoding human 5-lipoxygenase (5LO), was inserted into the vector and expressed in Saccharomyces cerevisiae. The enzyme (yh5LO) produced in the transformant was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A4 synthase activities, which were stimulated by Ca2+ and ATP. The N-terminal end of yh5LO contained five extra amino acids not present in 5LO purified from human leukocytes. A human 5LO-secretion vector containing the STA1 signal sequence was also constructed. When this hybrid gene was expressed in S. cerevisiae, its product was glycosylated and accumulated in the fractions related to the secretory pathway.

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Citations

Feb 1, 1994·Applied Biochemistry and Biotechnology·G BajszárJ R Mattoon
Oct 30, 1992·Biochimica Et Biophysica Acta·S Yamamoto
Oct 1, 1995·Journal of Lipid Mediators and Cell Signalling·O Rådmark
Nov 16, 2002·Prostaglandins & Other Lipid Mediators·Olof Rådmark
Jan 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·Y Y ZhangB Samuelsson
Dec 1, 1992·The Journal of Cell Biology·S KargmanJ F Evans
Feb 3, 2015·Critical Reviews in Biotechnology·Ruud HeshofJohanna Buchert
Dec 11, 1999·Proceedings of the Association of American Physicians·E S Silverman, J M Drazen
Oct 9, 2019·Journal of Agricultural and Food Chemistry·Holly StolterfohtHarald Pichler

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