title: Identification of new secreted proteins and secretion of heterologous amylase by C. glutamicum.
abstract: In this study, secreted Corynebacterium glutamicum proteins were investigated by two-dimensional gel electrophoresis. Around 100 spots observed in the pH range 4.5-5.5 had molecular masses that varied from 10 to 50 kDa. Upon N-terminal amino acid sequence analysis by Edman degradation, two of them were hits to two hypothetical proteins encoded by cgR_1176 and cgR_2070 on C. glutamicum R genome, respectively. Active-form alpha-amylase derived from Geobacillus stearothermophilus was successfully secreted by using the predicted cgR_1176 and cgR_2070 signal sequences, indicating that these hypothetical proteins were secreted proteins. Analysis using a disruption mutant of the twin-arginine translocation (Tat) export pathway machinery of C. glutamicum suggested that one is Tat pathway dependent secretion while the other is independent of the pathway. Our results demonstrate that C. glutamicum can secrete exoproteins by using its own signal sequences, indicating its potential as a host for protein productions.
authors.affiliation: Molecular Microbiology and Biotechnology Group, Research Institute of Innovative Technology for the Earth, 9-2, Kizugawadai, Kizugawa City, Kyoto, 619-0292, Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Suzuki
authors.forename: Keiro
authors.initials: K
authors.lastname: Watanabe
authors.forename: Naoko
authors.initials: N
authors.lastname: Okibe
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Tsuchida
authors.forename: Masayuki
authors.initials: M
authors.lastname: Inui
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yukawa
mesh_terms: D000595:Amino Acid Sequence; D000681:Amylases / Q000235:genetics / Q000378:metabolism*; D001406:Bacillaceae / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D021381:Protein Transport
chemical_list: D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D000681:Amylases