title: Characterization of lysine acetylation of a phosphoenolpyruvate carboxylase involved in glutamate overproduction in Corynebacterium glutamicum.
abstract: Protein Nε-acylation is emerging as a ubiquitous post-translational modification. In Corynebacterium glutamicum, which is utilized for industrial production of l-glutamate, the levels of protein acetylation and succinylation change drastically under the conditions that induce glutamate overproduction. Here, the acylation of phosphoenolpyruvate carboxylase (PEPC), an anaplerotic enzyme that supplies oxaloacetate for glutamate overproduction was characterized. It was shown that acetylation of PEPC at lysine 653 decreased enzymatic activity, leading to reduced glutamate production. An acetylation-mimic (KQ) mutant of K653 showed severely reduced glutamate production, while the corresponding KR mutant showed normal production levels. Using an acetyllysine-incorporated PEPC protein, we verified that K653-acetylation negatively regulates PEPC activity. In addition, NCgl0616, a sirtuin-type deacetylase, deacetylated K653-acetylated PEPC in vitro. Interestingly, the specific activity of PEPC was increased during glutamate overproduction, which was blocked by the K653R mutation or deletion of sirtuin-type deacetylase homologues. These findings suggested that deacetylation of K653 by NCgl0616 likely plays a role in the activation of PEPC, which maintains carbon flux under glutamate-producing conditions. PEPC deletion increased protein acetylation levels in cells under glutamate-producing conditions, supporting the hypothesis that PEPC is responsible for a large carbon flux change under glutamate-producing conditions.
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.|Kyowa Hakko Bio Co, Ltd., Chiyoda-ku, Tokyo, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Nagano-Shoji
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.|Kyowa Hakko Bio Co, Ltd., Chiyoda-ku, Tokyo, Japan.
authors.forename: Yuma
authors.initials: Y
authors.lastname: Hamamoto
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.|Kyowa Hakko Bio Co, Ltd., Chiyoda-ku, Tokyo, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Mizuno
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Ayuka
authors.initials: A
authors.lastname: Yamada
authors.affiliation: RIKEN Center for Life Science Technologies, Yokohama, Kanagawa, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kikuchi
authors.affiliation: RIKEN Center for Life Science Technologies, Yokohama, Kanagawa, Japan.
authors.forename: Mikako
authors.initials: M
authors.lastname: Shirouzu
authors.affiliation: RIKEN Center for Life Science Technologies, Yokohama, Kanagawa, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Umehara
authors.affiliation: RIKEN Center for Sustainable Resource Science, Wako, Saitama, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Nishiyama
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.|RIKEN Center for Sustainable Resource Science, Wako, Saitama, Japan.
authors.forename: Saori
authors.identifier: 0000-0001-7108-5142
authors.initials: S
authors.lastname: Kosono
mesh_terms: D000107:Acetylation; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011766:Pyruvate Carboxylase / Q000378:metabolism
chemical_list: D018698:Glutamic Acid; D010730:Phosphoenolpyruvate Carboxylase; D011766:Pyruvate Carboxylase; D008239:Lysine