Highly efficient export of a disulfide‐bonded protein to the periplasm and medium by the Tat pathway using CyDisCo in <i>Escherichia coli</i>

Arauzo-Aguilera, Klaudia; Saaranen, Mirva J.; Robinson, Colin; Ruddock, Lloyd W.

doi:10.1002/mbo3.1350

Cited by 7 publications

(5 citation statements)

References 32 publications

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“…Alternatively, the TAT pathway, another well-characterized secretion pathway, provides a fascinating alternative for protein secretion when proper folding occurred within the cytoplasm prior to secretion (Berks, 2015 ). Proteins such as GFP (Albiniak et al., 2013 ), protein glutaminase (Niu et al., 2019 ), disulfide bond‐containing protein (Arauzo-Aguilera et al., 2023 ), and laccases (Valimets et al., 2023 ) have been successfully secreted using the TAT pathway. Next, we will consider the possibility of NpAS being targeted for export via the TAT pathway.…”

Section: Resultsmentioning

confidence: 99%

Secretory expression of amylosucrase in Bacillus licheniformis through twin-arginine translocation pathway

Wang,

Niu,

Mchunu

et al. 2024

Journal of Industrial Microbiology and Biotechnology

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Amylosucrase (EC 2.4.1.4) is a versatile enzyme with significant potential in biotechnology and food production. To facilitate its efficient preparation, a novel expression strategy was implemented in Bacillus licheniformis for the secretory expression of Neisseria polysaccharea amylosucrase (NpAS). The host strain B. licheniformis CBBD302 underwent genetic modification through the deletion of sacB, a gene responsible for encoding levansucrase that synthesizes extracellular levan from sucrose, resulting in a levan-deficient strain, B. licheniformis CBBD302B. NpAS was successfully expressed in B. licheniformis CBBD302B using the highly efficient Sec-type signal peptide SamyL, but its extracellular translocation was unsuccessful. Consequently, the expression of NpAS via the twin-arginine translocation (TAT) pathway was investigated using the signal peptide SglmU. The study revealed that NpAS could be effectively translocated extracellularly through the TAT pathway, with the signal peptide SglmU facilitating the process. Remarkably, 62.81% of the total expressed activity was detected in the medium. This study marks the first successful secretory expression of NpAS in Bacillus species host cells, establishing a foundation for its future efficient production.

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Section: Resultsmentioning

confidence: 99%

Secretory expression of amylosucrase in Bacillus licheniformis through twin-arginine translocation pathway

Wang,

Niu,

Mchunu

et al. 2024

Journal of Industrial Microbiology and Biotechnology

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“…In this study, as one of the integral membrane proteins of Tat translocase, the expression of Sec-independent translocase protein TatA was differentially up-regulated (Figure S1 ) and enriched in the pathway of protein export and bacterial secretion. Arauzo-Aguilera et al (2023) reported recently protein YebF could be exported to the periplasm at high level through Tat pathway.…”

Section: Discussionmentioning

confidence: 99%

The Mechanism Insight into Bacterial Degradation of Pentachlorobiphenyl

Ji,

Chang,

Wang

et al. 2024

Preprint

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Bacterial degradation mechanism for high chlorinated pentachlorobiphenyl (PentaCB) with worse biodegradability has not been fully elucidated, which could limit the full remediation of PCBs-combined pollution. In this research, using enzymatic screening method, a new PentaCB-degrading bacterium M. paraoxydans that has not been reported was obtained. The characteristic of its intracellular enzymes, proteome and metabolome variation during PentaCB degradation were investigated systematically. The results showed that PentaCB (PCB101, 1 mg/L) degradation rate could arrive 23.9% within 4 h till complete degradation within 12 h. The intracellular enzyme compound was optimally active at pH 6.0. The 12 up-regulated characterized proteins involved ABC transporter substrate-binding protein, translocase protein TatA and signal peptidase I (SPase I) indicated that functional proteins for PentaCB degradation were present both in the cytoplasm and outer surface of cytoplasmic membrane. There were also 5 differential metabolites strongly associated with above proteins in which the up-regulated 1, 2, 4-benzenetriol was enriched into the degradation pathways of benzoate, chlorocyclohexane, chlorobenzene and aminobenzoate. Bacterial degradation of PentaCB necessitates transmembrane transport, energy consumption, protein export, biofilm formation and quorum sensing. These findings hold significant theory and application value for PCBs biodegradation.

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“…In this study, as one of the integral membrane proteins of Tat translocase, the expression of Sec-independent translocase protein TatA was differentially up-regulated ( Supplementary Figure S2 ) and enriched in the pathway of protein export and bacterial secretion. Arauzo-Aguilera et al ( 2023 ) reported recently protein YebF could be exported to the periplasm at high level through Tat pathway.…”

Section: Discussionmentioning

confidence: 99%

Insights into the biodegradation of pentachlorobiphenyl by Microbacterium paraoxydans: proteomic and metabolomic studies

Ji,

Chang,

Wang

et al. 2024

Front. Microbiol.

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Bacterial degradation mechanism for high chlorinated pentachlorobiphenyl (PentaCB) with worse biodegradability has not been fully elucidated, which could limit the full remediation of environments afflicted by the complex pollution of polychlorinated biphenyls (PCBs). In this research, a new PentaCB-degrading bacterium Microbacterium paraoxydans that has not been reported was obtained using enzymatic screening method. The characteristics of its intracellular enzymes, proteome and metabolome variation during PentaCB degradation were investigated systematically compared to non-PentaCB conditions. The findings indicate that the degradation rate of PentaCB (1 mg/L) could reach 23.9% within 4 hours and achieve complete degradation within 12 hours, with the mixture of intracellular enzymes being most effective at a pH of 6.0. During the biodegradation of PentaCB, the 12 up-regulated proteins characterized included ABC transporter PentaCB-binding protein, translocase protein TatA, and signal peptidase I (SPase I), indicating the presence of functional proteins for PentaCB degradation in both the cytoplasm and the outer surface of the cytoplasmic membrane. Furthermore, five differentially enriched metabolites were strongly associated with the aforementioned proteins, especially the up-regulated 1, 2, 4-benzenetriol which feeds into multiple degradation pathways of benzoate, chlorocyclohexane, chlorobenzene and aminobenzoate. These relevant results help to understand and speculate the complex mechanisms regarding PentaCB degradation by M. paraoxydans, which have both theoretical and practical implications for PCB bioremediation.

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Highly efficient export of a disulfide‐bonded protein to the periplasm and medium by the Tat pathway using CyDisCo in Escherichia coli

Cited by 7 publications

References 32 publications

Secretory expression of amylosucrase in Bacillus licheniformis through twin-arginine translocation pathway

Secretory expression of amylosucrase in Bacillus licheniformis through twin-arginine translocation pathway

The Mechanism Insight into Bacterial Degradation of Pentachlorobiphenyl

Insights into the biodegradation of pentachlorobiphenyl by Microbacterium paraoxydans: proteomic and metabolomic studies

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