Escherichia coli “TatExpress” strains export several g/L human growth hormone to the periplasm by the Tat pathway
Open accessAbstract
Escherichia coli is a heavily used platform for the production of biotherapeutic and other high‐value proteins, and a favored strategy is to export the protein of interest to the periplasm to simplify downstream processing and facilitate disulfide bond formation. The Sec pathway is the standard means of transporting the target protein but it is unable to transport complex or rapidly folding proteins because the Sec system can only transport proteins in an unfolded state. The Tat system also operates to transport proteins to the periplasm, and it has significant potential as an alternative means of recombinant protein production because it transports fully folded proteins. Here, we have tested the Tat system's full potential for the production of biotherapeutics for the first time using fed‐batch fermentation. We expressed human growth hormone (hGH) with a Tat signal peptide in E. coli W3110 “TatExpress” strains that contain elevated levels of the Tat apparatus. This construct contained four amino acids from TorA at the hGH N‐terminus as well as the initiation methionine from hGH, which is removed in vivo. We show that the protein is efficiently exported to the periplasm during extended fed‐batch fermentation, to the extent that it is by far the most abundant protein in the periplasm. The protein was shown to be homogeneous, disulfide bonded, and active. The bioassay showed that the yields of purified periplasmic hGH are 5.4 g/L culture whereas an enzyme‐linked immunosorbent assay gave a figure of 2.39 g/L. Separate analysis of a TorA signal peptide linked to hGH construct lacking any additional amino acids likewise showed efficient export to the periplasm, although yields were approximately two‐fold lower.
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The majority of recombinant proteins (>90%) translocating to the periplasm are directed via the SecB and SRP pathways.
“Although the TAT system has been successfully used in many cases [134] the majority of recombinant proteins (>90%) translocating to the periplasm are directed via the SecB and SRP pathways.”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [134]
The Introduction says that export to the periplasm is usually achieved through the Sec pathway, supporting the claim that Sec is commonly used. Our copy does not give a >90% figure or identify SecB and SRP as the pathways accounting for that proportion. (LLM, conf=0.86)
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Cites
Biopharmaceutical benchmarks 2014
Walsh, G. (2014) · Nature Biotechnology · doi:10.1038/nbt.3040 · Cited by 1,008 · see citing works
Natale, P., Brüser, T., & Driessen, A. J. M. (2007) · Biochimica et Biophysica Acta (BBA) - Biomembranes · doi:10.1016/j.bbamem.2007.07.015 · Cited by 543 · see citing works
Highly accurate genome sequences of Escherichia coli K‐12 strains MG1655 and W3110
Hayashi, K., Morooka, N., Yamamoto, Y., Fujita, K., Isono, K., Choi, S., Ohtsubo, E., Baba, T., Wanner, B. L., Mori, H., et al. (2006) · Molecular Systems Biology · doi:10.1038/msb4100049 · Cited by 503 · see citing works
Mode of Action of Pituitary Growth Hormone on Target Cells
Isaksson, O. G. P., Edén, S., & Jansson, J. (1985) · Annual Review of Physiology · doi:10.1146/annurev.ph.47.030185.002411 · Cited by 450 · see citing works
DeLisa, M. P., Tullman, D., & Georgiou, G. N. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0937838100 · Cited by 305 · see citing works
Patra, A. K., Mukhopadhyay, R., Mukhija, R., Krishnan, A., Garg, L. C., & Panda, A. K. (2000) · Protein Expression and Purification · doi:10.1006/prep.1999.1179 · Cited by 237 · see citing works
Advances in product release strategies and impact on bioprocess design
Balasundaram, B., Harrison, S., & Bracewell, D. G. (2009) · Trends in biotechnology · doi:10.1016/j.tibtech.2009.04.004 · Cited by 166 · see citing works
Purified human growth hormone from E. coli is biologically active
Olson, K. C., Fenno, J., Lin, N., Harkins, R. N., Snider, C. E., Kohr, W. H., Ross, M. J., Fodge, D. W., Prender, G., & STEBBING, N. (1981) · Nature · doi:10.1038/293408a0 · Cited by 133 · see citing works
Export Pathway Selectivity of Escherichia coli Twin Arginine Translocation Signal Peptides
Tullman‐Ercek, D., DeLisa, M. P., Kawarasaki, Y., Iranpour, P., Ribnicky, B., Palmer, T., & Georgiou, G. (2007) · Journal of Biological Chemistry · doi:10.1074/jbc.m610507200 · Cited by 129 · see citing works
The Crystal Structure of Affinity-matured Human Growth Hormone at 2 Å Resolution
Ultsch, M. H., Somers, W. S., Kossiakoff, A. A., & de Vos, A. M. (1994) · Journal of Molecular Biology · doi:10.1006/jmbi.1994.1135 · Cited by 115 · see citing works
Periplasmic production via the pET expression system of soluble, bioactive human growth hormone
Sockolosky, J. T., & Szoka, F. C. (2012) · Protein Expression and Purification · doi:10.1016/j.pep.2012.11.002 · Cited by 101 · see citing works
Salamone, D., Barañao, L., Santos, C. B., Bussmann, L. E., Artuso, J., Werning, C., Sterin‐Prync, A., Carbonetto, C., Dabsys, S., Munar, C. J., et al. (2006) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2006.01.005 · Cited by 83 · see citing works
Matos, C. F. R. O., Robinson, C., Alanen, H. I., Prus, P., Uchida, Y., Ruddock, L. W., Freedman, R. B., & Keshavarz‐Moore, E. (2013) · Biotechnology Progress · doi:10.1002/btpr.1858 · Cited by 55 · see citing works
Self-citationBrowning, D. F., Richards, K. L., Peswani, A. R., Roobol, J., Busby, S. J. W., & Robinson, C. (2017) · Biotechnology and Bioengineering · doi:10.1002/bit.26434 · Cited by 48 · see citing works
Self-citationMatos, C. F. R. O., Branston, S. D., Albiniak, A. M., Dhanoya, A., Freedman, R. B., Keshavarz‐Moore, E., & Robinson, C. (2012) · Biotechnology and Bioengineering · doi:10.1002/bit.24535 · Cited by 48 · see citing works
Self-citationAlanen, H. I., Walker, K. L., Velez‐Suberbie, M. L., Matos, C. F. R. O., Bönisch, S., Freedman, R. B., Keshavarz‐Moore, E., Ruddock, L. W., & Robinson, C. (2014) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/j.bbamcr.2014.12.027 · Cited by 45 · see citing works
Self-citationBiosynthesis and secretion of recombinant human growth hormone in Pichia pastoris
Ecamilla-Treviño, L. L., Viader-Salvadó, J. M., Barrera-Saldaña, H. A., & Guerrero‐Olazarán, M. (2000) · Biotechnology Letters · doi:10.1023/a:1005675920451 · Cited by 40 · see citing works
Humphreys, D. P., Vetterlein, O. M., Chapman, A. P., King, D. J., Antoniw, P., SUITTERS, A. J., Reeks, D. G., Parton, T. A. H., King, L. M., Smith, B. J., et al. (1998) · Journal of Immunological Methods · doi:10.1016/s0022-1759(98)00061-1 · Cited by 34 · see citing works
Self-citationOverall Protein Content and Induced Enzyme Components of the Periplasm of Bacillus subtilis
Pooley, H. M., Merchante, R., & Karamata, D. (1996) · Microbial Drug Resistance · doi:10.1089/mdr.1996.2.9 · Cited by 28 · see citing works
Safety of High Doses of Recombinant Human Growth Hormone
Van Loon, K. (1998) · Hormone Research in Paediatrics · doi:10.1159/000053092 · Cited by 23 · see citing works
Song, H., Jiang, J., Wang, X., & Zhang, J. (2016) · Bioengineered · doi:10.1080/21655979.2016.1212137 · Cited by 22 · see citing works
Hahm, M. S., & Chung, B. H. (2001) · Biotechnology and Bioprocess Engineering · doi:10.1007/bf02931995 · Cited by 17 · see citing works
Montero, I. G., Dolata, K. M., Schlüter, R., Malherbe, G., Sievers, S., Zühlke, D., Sura, T., Davé, E., Riedel, K., & Robinson, C. (2019) · Microbial Cell Factories · doi:10.1186/s12934-019-1071-7 · Cited by 16 · see citing works
Self-citation
Flagged first, then by citations
1 of the 24 references couldn’t be matched to a known paper.
Research works printed but not linked: 1
- Sambrook, J., & Russell, D. W. (2001). Molecular cloning: A laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
Cited by
Sorted by citations
Tripathi, N. K., & Shrivastava, A. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00420 · Cited by 470 · see citing works
Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments
Sandomenico, A., Sivaccumar, J. P., & Ruvo, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176324 · Cited by 134 · see citing works
Zhang, Z., Nong, F., Wang, Y., Yan, C.-X., Gu, Y. M., Song, P., & Sun, X. (2022) · Microbial Cell Factories · doi:10.1186/s12934-022-01917-y · Cited by 132 · see citing works
Microbial protein cell factories fight back?
Rettenbacher, L. A., Arauzo-Aguilera, K., Buscajoni, L., Castillo-Corujo, A., Ferrero-Bordera, B., Kostopoulou, A., Moran-Torres, R., Núñez-Nepomuceno, D., Öktem, A., Palma, A., et al. (2021) · Trends in Biotechnology · doi:10.1016/j.tibtech.2021.10.003 · Cited by 59 · see citing works
Self-citationCurrent trends in biopharmaceuticals production in Escherichia coli
McElwain, L., Phair, K., Kealey, C., & Brady, D. B. (2022) · Biotechnology Letters · doi:10.1007/s10529-022-03276-5 · Cited by 70 · see citing works
Niazi, S. K., & Magoola, M. (2023) · Biologics · doi:10.3390/biologics3040021 · Cited by 62 · see citing works
Strategies to Enhance Periplasmic Recombinant Protein Production Yields in Escherichia coli
Karyolaimos, A., & de Gier, J.-W. L. (2021) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2021.797334 · Cited by 45 · see citing works
Yoshida, K., & van Dijl, J. M. (2020) · Biotechnology and Bioprocess Engineering · doi:10.1007/s12257-020-0104-8 · Cited by 21 · see citing works
Zhang, Q., Chen, Y., Gao, L., Chen, J., Ma, X., Cai, D., Wang, D., & Chen, S. (2022) · Synthetic and Systems Biotechnology · doi:10.1016/j.synbio.2022.01.006 · Cited by 20 · see citing works
Zhu, J., Ruan, Y., Fu, X., Zhang, L.-C., Ge, G.-S., Wall, J., Zou, T., Zheng, Y., Ding, N., & Hu, X.-J. (2020) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.00313 · Cited by 19 · see citing works
Eskandari, A., Nezhad, N. G., Leow, T. C., Rahman, M. B. A., & Oslan, S. N. (2024) · Archives of Microbiology · doi:10.1007/s00203-024-03871-2 · Cited by 16 · see citing works
Taw, M. N., Li, M., Kim, D., Rocco, M. A., Waraho‐Zhmayev, D., & DeLisa, M. P. (2021) · ACS Synthetic Biology · doi:10.1021/acssynbio.1c00183 · Cited by 16 · see citing works
Schilling, T., Ferrero‐Bordera, B., Neef, J., Maaß, S., Becher, D., & van Dijl, J. M. (2023) · ACS Synthetic Biology · doi:10.1021/acssynbio.3c00444 · Cited by 14 · see citing works
Lin, Z., Ndengue, P. P. A., Jing, Y., Zhao, L., & Yang, X. (2021) · Protein Expression and Purification · doi:10.1016/j.pep.2021.105974 · Cited by 13 · see citing works
Arauzo‐Aguilera, K., Saaranen, M. J., Robinson, C., & Ruddock, L. W. (2023) · MicrobiologyOpen · doi:10.1002/mbo3.1350 · Cited by 12 · see citing works
Self-citationLin, Z., Jing, Y., Huang, Y., Yang, S., Chen, S., Ou, Y.-Y., Pistolozzi, M., & Yang, X. (2022) · Chemical Engineering Science · doi:10.1016/j.ces.2022.118052 · Cited by 11 · see citing works
Activation by NarL at the Escherichia coli ogt promoter
Ruanto, P., Chismon, D. L., Hothersall, J., Godfrey, R. E., Lee, D. J., Busby, S. J. W., & Browning, D. F. (2020) · Biochemical Journal · doi:10.1042/bcj20200408 · Cited by 11 · see citing works
Self-citationWang, Y., Kim, M., Buckley, C., Maynard, H. D., Langley, R. J., & Perry, J. K. (2023) · Protein Science · doi:10.1002/pro.4727 · Cited by 10 · see citing works
Mahalik, S., Sharma, A. K., Das, D., Mittra, D., & Mukherjee, K. J. (2022) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2022.04.012 · Cited by 5 · see citing works
Bacterial signal peptides: structure, optimization, and applications
Erkut, E. J. N. (2021) · Eureka · doi:10.29173/eureka28759 · Cited by 5 · see citing works
Biosensor that Detects Stress Caused by Periplasmic Proteins
Cumming, A. J., Khananisho, D., Bałka, M., Liljestrand, N., & Daley, D. O. (2024) · ACS Synthetic Biology · doi:10.1021/acssynbio.3c00720 · Cited by 3 · see citing works
Membrane and secretory protein production in Escherichia coli
Karyolaimos, A. (2021) · KTH Publication Database DiVA (KTH Royal Institute of Technology)
- Authors
- Guerrero Montero, I., Richards, K. L., Jawara, C., Browning, D. F., Peswani, A. R., Labrit, M., Allen, M., Aubry, C., Davé, E., Humphreys, D. P., Busby, S. J. W., & Robinson, C.
- Affiliations
- School of Biosciences University of Kent Canterbury United Kingdom
- University of Kent (GB)
- University of Birmingham (GB)
- UCB Pharma (United Kingdom) (GB)
- Corresponding author
- Robinson, Colin
- Venue
- Biotechnology and Bioengineering
- Publisher
- Wiley
- Published
- 2019-09-02
- Volume
- 116
- Issue
- 12
- Pages
- 3282-3291
- ISSN
- 0006-3592 (ISSN-L)
- 1097-0290
- PMC
- PMC6907408
- OpenAlex
- W2969640842
- Cited by
- 22
- References
- 24
- Citations by year
- 2024: 2 · 2023: 4 · 2022: 5 · 2021: 6 · 2020: 4 · 2019: 1
- FWCI
- 2.41
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Biotechnology and Bioengineering (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (acceptedVersion)
- Kent Academic Repository (University of Kent) (submittedVersion)
- Aston Publications Explorer (Aston University) (submittedVersion)
- University of Birmingham Research Portal (University of Birmingham) (submittedVersion, cc-by)
- Funders
- Biotechnology and Biological Sciences Research Council (BB/M018288/1, BB/P02789X/1)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- Tat pathway, human growth hormone, Tat signal peptide, fed-batch fermentation, recombinant protein expression, biologic therapy, disulfide bond formation, Escherichia coli W3110
- MeSH terms
- Escherichia coli, Humans, Recombinant Fusion Proteins, Protein Folding, Human Growth Hormone, Periplasm, Protein Sorting Signals
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