<scp>L</scp>-Glutamate Secretion by the N-Terminal Domain of the<i>Corynebacterium glutamicum</i>NCgl1221 Mechanosensitive Channel

Yamashita, Chikako; Hashimoto, Kana; Kumagai, Kosuke; Maeda, Tomoya; Takada, Ayako; Yabe, Isamu; Kawasaki, Hisashi; Wachi, Masaaki

doi:10.1271/bbb.120988

Cited by 26 publications

(18 citation statements)

References 37 publications

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“…Our previous study showed that the excessive biotin in corn stover completely blocked glutamic acid accumulation in the hydrolysate prepared from pretreated and detoxified corn stover [2]. In this study, we tried two metabolic modifications to facilitate glutamic acid secretion, either by restricting the biotin uptake to reduce the intracellular biotin content to a sub-optimal biotin level suitable for glutamate secretion [15], or by truncating the C-terminal amino acid residue to activate the activity of glutamate exporter MscCG [13, 14].…”

Section: Resultsmentioning

confidence: 99%

“…For glutamic acid secretion, the Ncgl1221 gene-encoded glutamate secretion channel protein MscCG was responsible for modulating glutamate export, and its C-terminal region was accountable for the gating process of the channel [13, 14]. The C-terminal 110 amino acid residue truncation achieved glutamate secretion in biotin-excessive conditions without inductions in two different C. glutamicum strains [13, 14]. Thus, we tried to truncate its C-terminal residues to activate glutamic acid secretion here.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, they were not suitable for glutamic acid fermentation in harsh inhibitors containing lignocellulose hydrolysate system. Glutamic acid secretion channel modification is also a potential way to achieve active glutamic acid secretion [13, 14], but its effect on inhibitors containing lignocellulose hydrolysate environment was still unclear. The metabolic engineering of C. glutamicum on triggering efficient glutamic acid accumulation from lignocellulose feedstock should balance the glutamic acid secretion from high-biotin-containing hydrolysate and the negative impact of the inhibitors in the hydrolysates on cell growth.…”

Section: Introductionmentioning

confidence: 99%

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Engineering Corynebacterium glutamicum triggers glutamic acid accumulation in biotin-rich corn stover hydrolysate

Wen

Bao

2019

Biotechnol Biofuels

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Background Lignocellulose biomass contains high amount of biotin and resulted in an excessive biotin condition for cellulosic glutamic acid accumulation by Corynebacterium glutamicum . Penicillin or ethambutol triggers cellulosic glutamic acid accumulation, but they are not suitable for practical use due to the fermentation instability and environmental concerns. Efficient glutamic acid production from lignocellulose feedstocks should be achieved without any chemical inductions. Results An industrial strain C. glutamicum S9114 was metabolically engineered to achieve efficient glutamic acid accumulation in biotin-excessive corn stover hydrolysate. Among the multiple metabolic engineering efforts, two pathway regulations effectively triggered the glutamic acid accumulation in lignocellulose hydrolysate. The C-terminal truncation of glutamate secretion channel MscCG (ΔC110) led to the successful glutamic acid secretion in corn stover hydrolysate without inductions. Then the α-oxoglutarate dehydrogenase complex (ODHC) activity was attenuated by regulating odhA RBS sequence, and glutamic acid accumulation was further elevated for more than fivefolds. The obtained C. glutamicum XW6 strain reached a record-high titer of 65.2 g/L with the overall yield of 0.63 g/g glucose using corn stover as the starting feedstock without any chemical induction. Conclusions Metabolic engineering method was successfully applied to achieve efficient glutamic acid in biotin-rich lignocellulose hydrolysate for the first time. This study demonstrated the high potential of glutamic acid production from lignocellulose feedstock. Electronic supplementary material The online version of this article (10.1186/s13068-019-1428-5) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Engineering Corynebacterium glutamicum triggers glutamic acid accumulation in biotin-rich corn stover hydrolysate

Wen

Bao

2019

Biotechnol Biofuels

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“…A recent study indicates that a loop which forms part of the cytoplasmic cage between residues 221-232 containing 3 negatively charged residues (IAPEILGELDVH) is essential for glutamate efflux as ablation of this loop abolishes glutamate efflux. 42 The exact role this loop plays in facilitating glutamate efflux is thus far unknown.…”

Section: Structural Determinants Of Mscs-like Channel Selectivitymentioning

confidence: 99%

Selectivity mechanisms in MscS-like channels

2013

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The E. coli mechanosensitive (MS) channel of small conductance (EcMscS) is the prototype of a diverse family of channels present in all domains of life. While EcMscS has been extensively studied, recent developments show that MscS may display some characteristics not widely conserved in this protein subfamily. With numerous members now electrophysiologically characterized, this subfamily of channels displays a breadth of ion selectivity with both anion and cation selective members. The selectivity of these channels may be relatively weak in comparison to voltage-gated channels but their selectivity mechanisms represent great novelty. Recent studies have identified unexpected residues important for selectivity in these homologs revealing different selectivity mechanisms than those employed by voltage gated K(+), Na(+), Ca(2+) and Cl(-) channels whose selectivity filters are housed within their transmembrane pores. This commentary looks at what is currently known about MscS subfamily selectivity and begins to unravel the potential physiological relevance of these differences.

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“…2) The importance of transporters responsible for substrate uptake and product release in fermentation that uses microorganisms is gradually being recognized, and studies paving the way for their technological utilization are currently underway. 3) The L-aspartate:L-alanine antiporter of Tetragenococcus halophilus (AspT) imports extracellular L-aspartate into the cell and releases intracellular L-alanine outside it. AspT acts in conjunction with aspartate decarboxylase (AspD) to create a proton concentration gradient and a membrane potential across the cell membrane, contributing to ATP synthesis by F1Fo-ATPase.…”

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confidence: 99%

R76 in transmembrane domain 3 of the aspartate:alanine transporter AspT is involved in substrate transport

Suzuki

Nanatani

Abe

2016

Bioscience, Biotechnology, and Biochemistry

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The L-aspartate:L-alanine antiporter of Tetragenococcus halophilus (AspT) possesses an arginine residue (R76) within the GxxxG motif in the central part of transmembrane domain 3 (TM3)-a residue that has been estimated to transport function. In this study, we carried out amino acid substitutions of R76 and used proteoliposome reconstitution for analyzing the transport function of each substitution. Both l-aspartate and l-alanine transport assays showed that R76K has higher activity than the AspT-WT (R76), whereas R76D and R76E have lower activity than the AspT-WT. These results suggest that R76 is involved in AspT substrate transport.

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L-Glutamate Secretion by the N-Terminal Domain of theCorynebacterium glutamicumNCgl1221 Mechanosensitive Channel

Cited by 26 publications

References 37 publications

Engineering Corynebacterium glutamicum triggers glutamic acid accumulation in biotin-rich corn stover hydrolysate

Engineering Corynebacterium glutamicum triggers glutamic acid accumulation in biotin-rich corn stover hydrolysate

Selectivity mechanisms in MscS-like channels

R76 in transmembrane domain 3 of the aspartate:alanine transporter AspT is involved in substrate transport

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