High-level expression of a sweet protein, monellin, in the food yeast Candida utilis.
We describe the heterologous expression of monellin in the yeast Candida utilis. A single-chain monellin gene was expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase [correction of decarboxylase] gene promoter from C. utilis. A promoter-deficient marker gene allowed high-copy-number integration of vectors into either the rDNA locus or the URA3 gene locus. Monellin was produced at a high level, accounting for > 50% of the soluble protein. No significant decrease in the production level of monellin was detected in transformants after 50 generations of nonselective growth.
Kondo
K
K
Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., Kanagawa, Japan. kondok@kirin.co.jp
Miura
Y
Y
Sone
H
H
Kobayashi
K
K
Iijima
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D001709:Biotechnology; D002175:Candida; D003001:Cloning, Molecular / Q000379:methods; D005819:Genetic Markers; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D013549:Sweetening Agents
D005819:Genetic Markers; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013549:Sweetening Agents; C002541:monellin protein, Dioscoreophyllum cumminsii; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases