Twin-Arginine Signal Peptide of <i>Bacillus subtilis</i> YwbN Can Direct Tat-Dependent Secretion of Methyl Parathion Hydrolase

Liu, Ruihua; Zuo, Zhenqiang; Xu, Yingming; Song, Chunyan; Jiang, Hong; Qiao, Chuanling; Xu, Ping; Zhou, Qixing; Yang, Chao

doi:10.1021/jf405694n

Cited by 16 publications

(10 citation statements)

References 31 publications

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“…Translocases TatA y C y and TatA d C d are responsible for secretion of YwbN and PhoD, respectively. As reported previously, phoD and ywbN SP were useful for secretion of heterologous proteins [28][29][30][31]. In this study, these four SPs were investigated for their effectiveness in secretion of recombinant GehC.…”

Section: Construction Of Gehc Vectors and Various Lipase-producing Stmentioning

confidence: 81%

Strategy for efficient production of recombinant Staphylococcus epidermidis lipase in Bacillus subtilis

Chen

Lin

et al. 2015

Biochemical Engineering Journal

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Section: Construction Of Gehc Vectors and Various Lipase-producing Stmentioning

confidence: 81%

Strategy for efficient production of recombinant Staphylococcus epidermidis lipase in Bacillus subtilis

Chen

Lin

et al. 2015

Biochemical Engineering Journal

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“…There are several well-characterized secretion pathways elucidated in B. subtilis , especially the general secretory (Sec) pathway and the twin-arginine (Tat) translocation pathway. We selected four signal peptides (SP amyL , SP lipA , SP nprB , and SP nprE ) from Sec pathway − and two (SP phoD and SP ywbN ) from Tat pathway, , which were reported to highly direct the secretion of different enzymes.…”

Section: Resultsmentioning

confidence: 99%

High-Efficiency Secretion of β-Mannanase in Bacillus subtilis through Protein Synthesis and Secretion Optimization

Song

Dong

et al. 2017

J. Agric. Food Chem.

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The manno endo-1,4-mannosidase (β-mannanase, EC. 3.2.1.78) catalyzes the random hydrolysis of internal (1 → 4)-β-mannosidic linkages in the mannan polymers. A codon optimized β-mannanase gene from Bacillus licheniformis DSM13 was expressed in Bacillus subtilis. When four Sec-dependent and two Tat-dependent signal peptide sequences cloned from B. subtilis were placed upstream of the target gene, the highest activity of β-mannanase was observed using SP as a signal peptide. Then a 1.25-fold activity of β-mannanase was obtained when another copy of groESL operon was inserted into the genome of host strain. Finally, five different promoters were separately used to enhance the synthesis of the target protein. The results showed that promoter P, a modified maltose-inducible promoter, significantly elevated the production of β-mannanase. After 72 h of flask fermentation, the enzyme activity of β-mannanase in the supernatant when using locust bean gum as substrate reached 2207 U/mL. This work provided a promising β-mannanase production strain in industrial application.

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“…Still other proteins are exported into the medium via ATP-binding cassette (ABC) transporters [ 12 ]. With the development of investigation involved in Sec and Tat pathway, many Sec-dependent [ 13 – 15 ] and Tat-dependent [ 16 , 17 ] signal peptides were applied in secretion of proteins of interest. However, the high-level secretion of heterologous proteins with signal peptides is unfortunately not that straight forward.…”

Section: Introductionmentioning

confidence: 99%

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

Chen

Zhao

et al. 2016

Microb Cell Fact

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BackgroundThe Gram-positive bacterium Bacillus subtilis has been widely used as a cell factory for the production of proteins due to its generally regarded as safe (GRAS) nature and secretion capability. Of the known secretory pathways in B. subtilis, the majority of proteins are exported from the cytoplasm by Sec pathway, Tat pathway and ABC transporters, etc. However, the production of heterologous proteins by B. subtilis is unfortunately not that straight forward because of the bottlenecks in classical secretion pathways. The aim of this work is to explore a new method for protein production based on non-classical secretion pathway.ResultsOne d-psicose 3-epimerase (RDPE) which converts d-fructose into d-psicose from Ruminococcus sp. 5_1_39BFAA was successfully and substantially secreted into the extracellular milieu without the direction of signal peptide. Subsequently, we demonstrated that RDPE contained no native signal peptide, and the secretion of RDPE was not dependent on Sec or Tat pathway or due to cell lysis, which indicated that RDPE is a non-classically secreted protein. Then, we attempted to evaluate the possibility of using RDPE as a signal to export eighteen reporter proteins into the culture medium. Five of eleven homologous proteins, two of five heterologous proteins from other bacterium and two heterologous proteins of eukaryotic source were successfully secreted into the extracellular milieu at different secretion levels when they were fused to RDPE mediated by a flexible 21-bp linker to keep a distance between two single proteins. Furthermore, the secretion rates of two fusion proteins (RDPE-DnaK and RDPE-RFP) reached more than 50 %. In addition, most of the fusion proteins retained enzyme or biological activity of their corresponding target proteins, and all of the fusions still had the activity of RDPE.ConclusionsWe found and identified a heterologous non-classically secreted protein RDPE, and showed that RDPE could direct proteins of various types into the culture medium, and thus non-classical protein secretion pathway can be used as a novel secretion pathway for recombinant proteins. This novel strategy for recombinant protein production is helpful to make B. subtilis as a more ideal cell factory for protein production.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0469-8) contains supplementary material, which is available to authorized users.

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Twin-Arginine Signal Peptide of Bacillus subtilis YwbN Can Direct Tat-Dependent Secretion of Methyl Parathion Hydrolase

Cited by 16 publications

References 31 publications

Strategy for efficient production of recombinant Staphylococcus epidermidis lipase in Bacillus subtilis

Strategy for efficient production of recombinant Staphylococcus epidermidis lipase in Bacillus subtilis

High-Efficiency Secretion of β-Mannanase in Bacillus subtilis through Protein Synthesis and Secretion Optimization

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

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