High-yield production of L-serine through a novel identified exporter combined with synthetic pathway in Corynebacterium glutamicum

Zhang, Xiaomei; Gao, Yujie; Chen, Ziwei; Xu, Guowen; Zhang, Xiaojuan; Li, Hui; Shi, Jin‐Song; Koffas, Mattheos; Xu, Zhenghong

doi:10.21203/rs.3.rs-16998/v2

Cited by 4 publications

(9 citation statements)

References 24 publications

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“…By employing a deubiquitination strategy, the accumulation of L-malic acid was improved in S. cerevisiae [50]. In another case, to export L-serine from the cell, a novel exporter SerE was overexpressed, and the titer of L-serine reached 43.9 g/L in C. glutamicum combining with the strengthening of Lserine synthetic pathway, which further enhanced its industrial application [51].…”

Section: Improvement Of the Transmembrane Export Of Target Products I...mentioning

confidence: 99%

Transporter Engineering in Microbial Cell Factory Boosts Biomanufacturing Capacity

Xue

Qin

et al. 2022

BioDesign Res

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Microbial cell factories (MCFs) are typical and widely used platforms in biomanufacturing for designing and constructing synthesis pathways of target compounds in microorganisms. In MCFs, transporter engineering is especially significant for improving the biomanufacturing efficiency and capacity through enhancing substrate absorption, promoting intracellular mass transfer of intermediate metabolites, and improving transmembrane export of target products. This review discusses the current methods and strategies of mining and characterizing suitable transporters and presents the cases of transporter engineering in the production of various chemicals in MCFs.

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Section: Improvement Of the Transmembrane Export Of Target Products I...mentioning

confidence: 99%

Transporter Engineering in Microbial Cell Factory Boosts Biomanufacturing Capacity

Xue

Qin

et al. 2022

BioDesign Res

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“…The strains F343-BS, -CS, -AS, -S, -B, -C, and -A were grown in the fermentation medium for 24 h, and the cells were harvest, washed twice with PBS (pH 7.4) [39], and visualized using a Carl Zeiss LSM880 microscope (Carl Zeiss Microscopy GmbH, Jena, Germany) equipped with fully automatic inverted microscope and 63×/1.40 oil plan apochromatic objective lens. The excitation lter was 488 nm and emission lter was 510-550 nm.…”

Section: Confocal Microscopic Observationmentioning

confidence: 99%

Functional Caracterization of CapBCA in Controlling Poly-γ-Glutamic Acid Synthesis in Corynebacterium Glutamicum

Wang

et al. 2021

Preprint

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Background Poly-γ-glutamic acid (γ-PGA) is a natural anionic biopolymer widely used in various fields, including medicine, food, cosmetics, and environmental protection. The γ-PGA synthase complex, CapBCA, is the only polyprotein complex responsible for γ-PGA synthesis. However, systematic and in-depth research on the function of each component involved in γ-PGA synthesis is scarce, which limits enhanced production of γ-PGA. Results To address this limitation, γ-PGA synthase components were localized, and their functions associated with γ-PGA synthesis were investigated in Corynebacterium glutamicum. Bioinformatics analysis and confocal microscopic observations of CapB-sfGFP, CapC-sfGFP, and CapA-sfGFP proteins revealed that γ-PGA synthase components CapB, CapC, and CapA were all localized on the cell membrane. More importantly, γ-PGA was detected only when CapB, CapC, and CapA were expressed in combination in C. glutamicum. Furthermore, enhancement of CapB or CapC transcription levels (from low to high) and maintaining medium-level CapA transcription led to 35.44% and 76.53% increase in γ-PGA yield (γ-PGA yield-to-biomass), respectively. However, maintaining medium-level CapB and CapC transcription, and moderate enhancement of CapA transcription level (from low to medium) led to 35.01% increase in γ-PGA yield, whereas a further increase in CapA expression (from medium to high) led to 10.36% decrease in γ-PGA yield. Notably, CapC had the greatest influence (accounting for 68.24%) on γ-PGA synthesis. Conclusions The present study determined the membrane localization of γ-PGA synthase components, CapB, CapC, and CapA, in C. glutamicum and confirmed the significance of these components in γ-PGA production. Furthermore, CapC was found to have the greatest influence on controlling γ-PGA synthesis. These findings shed light into the effect of γ-PGA synthase component expression on γ-PGA synthesis, and provide insights for further improvement in γ-PGA production.

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“…The copyright holder for this preprint this version posted July 18, 2022. ; https://doi.org/10.1101/2022.07. 13.499997 doi: bioRxiv preprint transcriptomics and differential gene functional analyses of four strains with different L-serine titers (SYPS-062, 6.6 g/L; SYPS-062-33a, 13.2 g/L; SSAAI, 26 g/L; A36, 31 g/L) (19,(23)(24)(25)(26). Next, the contribution of each exporter to L-serine export was studied by gene deletion, gene overexpression and amino acid export assay.…”

Section: Introductionmentioning

confidence: 99%

“…4 In the past few years, only seven amino acid exporters have been identified in C. glutamicum, involved in the export of 13 amino acids (Fig. 1) (11)(12)(13)(14)(15)(16)(17)(18)(19)(20). Among the seven exporters, BrnFE participates in the export of L-methionine, L-isoleucine, L-valine, and L-leucine (11,12).…”

Section: Introductionmentioning

confidence: 99%

Novel insights into L-serine exporters in Corynebacterium glutamicum from gene mining and functional analysis

Gao

Zhang

et al. 2022

Preprint

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Amino acid exporters play an important role in regulating amino acid production by Corynebacterium glutamicum, and over 90% of amino acid export is attributed to exporters in this species. ThrE was reported to be an L-serine exporter, and SerE was identified as an L-serine exporter in our previous study. However, when both ThrE and SerE were deleted, the L-serine titer was decreased by 60%, suggesting other L-serine exporters may exist. In the present study, NCgl0254 and NCgl0255 were identified as novel L-serine exporters through comparative transcriptomics and gene functional analyses. The contributions of the four exporters (ThrE, SerE, NCgl0254 and NCgl0255) in L-serine export were studied by gene deletion, gene overexpression, amino acid export assay and real-time quantitative PCR (RT-qPCR). The results showed that SerE is the major L-serine exporter in C. glutamicum. Fermentation and amino acid export assays of SSAAI, SSAAI-serE-thrE-ncgl0254-ncgl0255 and SSAAIΔserEΔthrEΔncgl0254Δncgl0255 indicated that the four L-serine exporters undertake most of the L-serine export, and their overexpression enhanced export of L-serine in SSAAI. When one L-serine exporter was deleted, the transcription level of the other three exporters was upregulated. However, the decrease in L-serine titer caused by deletion of one exporter was not fully compensated by upregulation of the other three exporters at the transcription level, indicating that L-serine production by C. glutamicum may be determined by cooperative efficiency of all four L-serine exporters, with each being interdependent.

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High-yield production of L-serine through a novel identified exporter combined with synthetic pathway in Corynebacterium glutamicum

Cited by 4 publications

References 24 publications

Transporter Engineering in Microbial Cell Factory Boosts Biomanufacturing Capacity

Transporter Engineering in Microbial Cell Factory Boosts Biomanufacturing Capacity

Functional Caracterization of CapBCA in Controlling Poly-γ-Glutamic Acid Synthesis in Corynebacterium Glutamicum

Novel insights into L-serine exporters in Corynebacterium glutamicum from gene mining and functional analysis

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