title: Altered acetylation and succinylation profiles in Corynebacterium glutamicum in response to conditions inducing glutamate overproduction.
abstract: The bacterium Corynebacterium glutamicum is utilized during industrial fermentation to produce amino acids such as L-glutamate. During L-glutamate fermentation, C. glutamicum changes the flux of central carbon metabolism to favor L-glutamate production, but the molecular mechanisms that explain these flux changes remain largely unknown. Here, we found that the profiles of two major lysine acyl modifications were significantly altered upon glutamate overproduction in C. glutamicum; acetylation decreased, whereas succinylation increased. A label-free semi-quantitative proteomic analysis identified 604 acetylated proteins with 1328 unique acetylation sites and 288 succinylated proteins with 651 unique succinylation sites. Acetylation and succinylation targeted enzymes in central carbon metabolic pathways that are directly related to glutamate production, including the 2-oxoglutarate dehydrogenase complex (ODHC), a key enzyme regulating glutamate overproduction. Structural mapping revealed that several critical lysine residues in the ODHC components were susceptible to acetylation and succinylation. Furthermore, induction of glutamate production was associated with changes in the extent of acetylation and succinylation of lysine, suggesting that these modifications may affect the activity of enzymes involved in glutamate production. Deletion of phosphotransacetylase decreased the extent of protein acetylation in nonproducing condition, suggesting that acetyl phosphate-dependent acetylation is active in C. glutamicum. However, no effect was observed on the profiles of acetylation and succinylation in glutamate-producing condition upon disruption of acetyl phosphate metabolism or deacetylase homologs. It was considered likely that the reduced acetylation in glutamate-producing condition may reflect metabolic states where the flux through acid-producing pathways is very low, and substrates for acetylation do not accumulate in the cell. Succinylation would occur more easily than acetylation in such conditions where the substrates for both acetylation and succinylation are limited. This is the first study investigating the acetylome and succinylome of C. glutamicum, and it provides new insight into the roles of acyl modifications in C. glutamicum biology.
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.|Kyowa Hakko Bio Co., Ltd., Tokyo, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Mizuno
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.|Kyowa Hakko Bio Co., Ltd., Tokyo, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Nagano-Shoji
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.|Department of Environmental Materials Science, Tokyo Denki University, Tokyo, Japan.
authors.forename: Shosei
authors.initials: S
authors.lastname: Kubo
authors.affiliation: RIKEN Center for Sustainable Resource Science, Saitama, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kawamura
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Yoshida
authors.affiliation: Department of Environmental Materials Science, Tokyo Denki University, Tokyo, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Kawasaki
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Nishiyama
authors.affiliation: RIKEN Center for Sustainable Resource Science, Saitama, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Yoshida
authors.affiliation: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.|RIKEN Center for Sustainable Resource Science, Saitama, Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Kosono
mesh_terms: D000107:Acetylation; D000595:Amino Acid Sequence; D019149:Bioreactors / Q000382:microbiology; D048230:Corynebacterium glutamicum / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018698:Glutamic Acid / Q000096:biosynthesis*; D008239:Lysine / Q000378:metabolism; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways / Q000502:physiology*; D010709:Phosphate Acetyltransferase / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D020543:Proteome / Q000032:analysis
chemical_list: D020543:Proteome; D018698:Glutamic Acid; D010709:Phosphate Acetyltransferase; D008239:Lysine
keywords: 2-oxoglutarate dehydrogenase complex; corynebacterium glutamicum; l-glutamate overproduction; label-free semi-quantitative proteomic analysis; lysine acetylation; lysine succinylation.