Efficient expression of bovine alpha-lactalbumin in Saccharomyces cerevisiae.
A synthetic gene encoding the mature bovine alpha-lactalbumin fused to the preproregion of the yeast alpha-mating factor has been expressed and secreted at high level in Saccharomyces cerevisiae under the control of the alpha-mating promoter. Growth conditions were found to be critical for the expression: recombinant alpha-lactalbumin could only be detected in the medium provided the culture was grown at neutral pH. The secreted bovine alpha-lactalbumin is enzymatically active and identical to the whey protein, as confirmed by SDS/PAGE, IEF, ultraviolet and CD spectral analysis, and amino-terminal sequence determination.
Viaene
A
A
Interdisciplinary Research Center, Katholieke Universiteit Leuven, Belgium.
Volckaert
G
G
Joniau
M
M
De Baetselier
A
A
Van Cauwelaert
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005813:Genes, Synthetic; D007768:Lactalbumin / Q000235:genetics* / Q000302:isolation & purification; D007785:Lactose / Q000096:biosynthesis; D008969:Molecular Sequence Data; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D011994:Recombinant Proteins; D004247:DNA; D007768:Lactalbumin; D007785:Lactose