Expression and secretion of a single-chain sweet protein, monellin, in Saccharomyces cerevisiae by an α-factor signal peptide.
The sweet protein monellin gene was expressed in Saccharomyces cerevisiae under the control of the GAL1 promoter and α-factor signal peptide sequence of S. cerevisiae. The gene, which was obtained through mutation of the synthesized single-chain monellin gene, was cloned into an E. coli-yeast shuttle vector pYES2.0 which carries the galactose-inducible promoter GAL1. Then the α-factor signal peptide of S. cerevisiae was linked also, resulting in the secreting expression vector pYESMTA. The recombinant plasmid was subsequently transformed into strain S. cerevisiae INVsc1. The peptide efficiently directed the secretion of monellin from the recombinant yeast cell. A maximum yield of active monellin was 0.41 g l(-1) of the supernatant from INVsc1 harboring pYESMTA.
College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot, People's Republic of China. chenzhongjun2001@yahoo.com.cn
Zhongjun
Z
Chen
Zhengying
Z
Li
Na
N
Yu
Lili
L
Yan
D000595:Amino Acid Sequence; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D013549:Sweetening Agents / Q000378:metabolism
D010940:Plant Proteins; D021382:Protein Sorting Signals; D013549:Sweetening Agents; C069034:monellin, Asn(22)-Gln(25)-Asn(26)-A-chain-Asn(49)-Glu(50)-B-chain-