title: Mutagenesis of the glucoamylase signal peptide of Saccharomyces diastaticus and functional analysis in Saccharomyces cerevisiae.
abstract: To improve the efficiency of the glucoamylase signal peptide (GSP) of Saccharomyces diastaticus for the secretion of foreign proteins, hybrid plasmids containing one of four types of GSP mutant (m1, Pro(-18)-->Leu(-18); m2, Tyr(-13)-->Leu(-13); m3, Ser(-9)-->Leu(-9); m4, Asn(-5)-->Pro(-5)) were constructed and evaluated in Saccharomyces cerevisiae using Bacillus endo-1,4-beta-D-glucanase (CMCase) as a reporter gene. CMCase secretion by m1, m2 and m3 GSP mutants was increased, likely resulting from a higher probability of the modified GSP to assume an alpha-helical structure. Especially in the case of m3, the substitution of Leu for a polar residue, Ser(-9), in the hydrophobic region resulted in approximately a twofold increase in extracellular CMCase activity. In mutant 4, which disrupts the alpha-helix of GSP, CMCase was less efficiently secreted.
authors.affiliation: Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejin University, Yusong, Taejon, South Korea.
authors.forename: J W
authors.initials: JW
authors.lastname: Lee
authors.forename: D O
authors.initials: DO
authors.lastname: Kang
authors.forename: B Y
authors.initials: BY
authors.lastname: Kim
authors.forename: W K
authors.initials: WK
authors.lastname: Oh
authors.forename: T I
authors.initials: TI
authors.lastname: Mheen
authors.forename: Y R
authors.initials: YR
authors.lastname: Pyun
authors.forename: J S
authors.initials: JS
authors.lastname: Ahn
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002480:Cellulase / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D021382:Protein Sorting Signals / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D012440:Saccharomyces / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D005087:Glucan 1,4-alpha-Glucosidase; D002480:Cellulase; C019826:carboxymethylcellulase