An efficient protocol to enhance the extracellular production of recombinant protein from Escherichia coli by the synergistic effects of sucrose, glycine, and Triton X-100
Abstract
Targeting recombinant proteins at highly extracellular production in the culture medium of Escherichia coli presents a significant advantage over cytoplasmic or periplasmic expression. In this work, a recombinant protein between ZZ protein and alkaline phosphatase (rZZ-AP) was constructed. Because rZZ-AP has the IgG-binding capacity and enzymatic activity, it can serve as an immunoreagent in immunoassays. However, only a very small portion of rZZ-AP is generally secreted into the aqueous medium under conventional cultivation procedure. Hence, we emphasized on the optimization of the culture procedures and attempted to dramatically enhance the yield of extracellular rZZ-AP from E. coli HB101 host cells by adding sucrose, glycine, and Triton X-100 in the culture medium. Results showed that the extracellular production of rZZ-AP in the culture medium containing 5% sucrose, 1% glycine, and 1% Triton X-100 was 18.6 mg/l, which was 18.6-fold higher than that without the three chemicals. And the β-galactosidase activity test showed that the increased extracellular rZZ-AP was not due to cell lysis. Further analysis suggested a significant interaction effect among the three chemicals for the enhancement of extracellular production. Ultrastructural analysis indicated that the enhancement may be due to the influence of sucrose, glycine, and Triton X-100 on the periplasmic osmolality, permeability, or integrity of the cell wall, respectively. This proposed approach presents a simple stra…
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Glycine induces swelling of E. coli cells and enlargement of the periplasmic space by interfering with peptidoglycan synthesis, while Triton X-100 disrupts the integrity of the outer membrane.
“Indeed, while glycine induces the swelling of E. coli cells and enlargement of the periplasmic space by interfering with the synthesis of peptidoglycans, Triton X-100 disrupts the integrity of the outer membrane [174].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [174]
Cites
Recombinant protein expression in Escherichia coli: advances and challenges
Rosano, G. L., & Ceccarelli, E. A. (2014) · Frontiers in Microbiology · doi:10.3389/fmicb.2014.00172 · Cited by 2,556 · see citing works
Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay (ELISA)
Lequin, R. M. (2005) · Clinical Chemistry · doi:10.1373/clinchem.2005.051532 · Cited by 1,562 · see citing works
Recombinant protein expression in Escherichia coli
Baneyx, F. (1999) · Current Opinion in Biotechnology · doi:10.1016/s0958-1669(99)00003-8 · Cited by 1,485 · see citing works
Advanced genetic strategies for recombinant protein expression in Escherichia coli
Sørensen, H. P., & Mortensen, K. K. (2004) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2004.08.004 · Cited by 971 · see citing works
Yamanaka, M., Hara, K., & Kudo, J. (2005) · Applied and Environmental Microbiology · doi:10.1128/aem.71.11.7589-7593.2005 · Cited by 907 · see citing works
Multiple Mechanisms of Protein Insertion into and Across Membranes
Wickner, W. T., & Lodish, H. F. (1985) · Science · doi:10.1126/science.4048938 · Cited by 698 · see citing works
Secretory and extracellular production of recombinant proteins using Escherichia coli
Choi, J. H., & Lee, S. Y. (2004) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-004-1559-9 · Cited by 614 · see citing works
Recombinant protein secretion in Escherichia coli
Mergulhão, F. J. M., Summers, D. K., & Monteiro, G. A. (2005) · Biotechnology Advances · doi:10.1016/j.biotechadv.2004.11.003 · Cited by 485 · see citing works
Outer membrane of gram-negative bacteria. XII. Molecular-sieving function of cell wall
Decad, G. M., & Nikaido, H. (1976) · Journal of Bacteriology · doi:10.1128/jb.128.1.325-336.1976 · Cited by 299 · see citing works
Chapter 17 Refolding Solubilized Inclusion Body Proteins
Burgess, R. R. (2009) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(09)63017-2 · Cited by 254 · see citing works
High level production of soluble single chain antibodies in small-scale Escherichia coli cultures
Kipriyanov, S. M., Moldenhauer, G., & Little, M. (1997) · Journal of Immunological Methods · doi:10.1016/s0022-1759(96)00188-3 · Cited by 201 · see citing works
The Periplasmic Escherichia coli Peptidylprolyl cis,trans-Isomerase FkpA
Bothmann, H., & Plückthun, A. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m910233199 · Cited by 192 · see citing works
Bach, H., Mazor, Y., Shaky, S., Shoham-Lev, A., Berdichevsky, Y., Gutnick, D. L., & Benhar, I. (2001) · Journal of Molecular Biology · doi:10.1006/jmbi.2001.4914 · Cited by 178 · see citing works
Secretory Production of Recombinant Proteins in Escherichia coli
Yoon, S. H., Kim, S., & Kim, J. (2009) · Recent Patents on Biotechnology · doi:10.2174/187220810790069550 · Cited by 145 · see citing works
Moks, T., Abrahmsén, L., Holmgren, E., Bilich, M., Olsson, A., Pohl, G., Sterky, C., Hultberg, H., & Josephson, S. (1987) · Biochemistry · doi:10.1021/bi00391a005 · Cited by 138 · see citing works
Optimisation of signal peptide for recombinant protein secretion in bacterial hosts
Low, K. O., Muhammad Mahadi, N., & Md. Illias, R. (2013) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-013-4831-z · Cited by 130 · see citing works
Folding and aggregation of beta-lactamase in the periplasmic space of Escherichia coli.
Bowden, G. A., & Georgiou, G. (1990) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)44825-3 · Cited by 125 · see citing works
Elasticity of the sacculus of Escherichia coli
Koch, A. L., & Woeste, S. (1992) · Journal of Bacteriology · doi:10.1128/jb.174.14.4811-4819.1992 · Cited by 120 · see citing works
Simmons, L. C., & Yansura, D. G. (1996) · Nature Biotechnology · doi:10.1038/nbt0596-629 · Cited by 110 · see citing works
Technical refolding of proteins: Do we have freedom to operate?
Eiberle, M. K., & Jungbauer, A. (2010) · Biotechnology Journal · doi:10.1002/biot.201000001 · Cited by 108 · see citing works
Secretion of heterologous gene products to the culture medium ofEscherichia coli
Abrahmsén, L., Moks, T., Nilsson, B., & Uhlén, M. (1986) · Nucleic Acids Research · doi:10.1093/nar/14.18.7487 · Cited by 103 · see citing works
Hwang, P. M., Pan, J., & Sykes, B. D. (2013) · FEBS Letters · doi:10.1016/j.febslet.2013.09.028 · Cited by 100 · see citing works
Refolding of proteins from inclusion bodies: rational design and recipes
Basu, A., Li, X., & Leong, S. S. J. (2011) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-011-3513-y · Cited by 91 · see citing works
An efficient protocol to enhance recombinant protein expression using ethanol in Escherichia coli
Chhetri, G., Kalita, P., & Tripathi, T. (2015) · MethodsX · doi:10.1016/j.mex.2015.09.005 · Cited by 86 · see citing works
Extracellular recombinant protein production from an Escherichia coli lpp deletion mutant
Shin, H., & Chen, R. R. (2008) · Biotechnology and Bioengineering · doi:10.1002/bit.22013 · Cited by 86 · see citing works
A family of arabinose-inducible Escherichia coli expression vectors having pBR322 copy control
Cronan, J. E. (2005) · Plasmid · doi:10.1016/j.plasmid.2005.07.001 · Cited by 85 · see citing works
Extracellular production of recombinant proteins using bacterial autotransporters
Jong, W. S. P., Saurı́, A., & Luirink, J. (2010) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2010.07.009 · Cited by 78 · see citing works
Yang, J., Moyana, T. N., MacKenzie, S. L., Xia, Q., & Xiang, J. (1998) · Applied and Environmental Microbiology · doi:10.1128/aem.64.8.2869-2874.1998 · Cited by 75 · see citing works
Jonet, M. A., Mahadi, N. M., Murad, A. M. A., Rabu, A., Bakar, F. D. A., Rahim, R. A., Low, K. O., & Illias, R. M. (2012) · Microbial Physiology · doi:10.1159/000336524 · Cited by 65 · see citing works
Li, Z., Gu, Z., Wang, M., Du, G., Wu, J., & Chen, J. (2009) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-009-2157-7 · Cited by 51 · see citing works
Immunodiagnostic reagents using llama single domain antibody–alkaline phosphatase fusion proteins
Swain, M. D., Anderson, G. P., Serrano-González, J., Liu, J. L., Zabetakis, D., & Goldman, E. R. (2011) · Analytical Biochemistry · doi:10.1016/j.ab.2011.06.012 · Cited by 41 · see citing works
pDAP2: a vector for construction of alkaline phosphatase fusion-proteins
Kerschbaumer, R. J., Hirschl, S., Schwager, C., Ibl, M., & Himmler, G. (1996) · Immunotechnology · doi:10.1016/1380-2933(96)00040-1 · Cited by 40 · see citing works
Accessing genomic information: alternatives to PCR
Isaksson, A., & Landegren, U. D. (1999) · Current Opinion in Biotechnology · doi:10.1016/s0958-1669(99)80003-2 · Cited by 34 · see citing works
A novel strategy for enhancing extracellular secretion of recombinant proteins in Escherichia coli
Su, L., Xu, C., Woodard, R. W., Chen, J., & Wu, J. (2013) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-013-4994-7 · Cited by 33 · see citing works
Rau, D., Kramer, K., & Hock, B. (2002) · Journal of Immunoassay and Immunochemistry · doi:10.1081/ias-120003657 · Cited by 30 · see citing works
Grigorenko, V., Andreeva, I. P., Börchers, T., Spener, F., & Egorov, A. M. (2001) · Analytical Chemistry · doi:10.1021/ac000684t · Cited by 28 · see citing works
Martı́nez, A., Ramı́rez, O. T., & Valle, F. (1997) · Applied Microbiology and Biotechnology · doi:10.1007/s002530050885 · Cited by 25 · see citing works
Effects of glycine supplement on protein production and release in recombinant Escherichia coli
Aristidou, A. A., Yu, P., & San, K. (1993) · Biotechnology Letters · doi:10.1007/bf00128271 · Cited by 25 · see citing works
Effect of Glycine and Triton X-100 on secretion and expression of ZZ–EGFP fusion protein
Tang, J., Yang, H., Song, S., Zhu, P., & Ji, A. (2007) · Food Chemistry · doi:10.1016/j.foodchem.2007.11.034 · Cited by 22 · see citing works
Self-citationGenetically Fused Protein A–Luciferase for Immunological Blotting Analyses
Zhang, X., Kobatake, E., Kobayashi, K., Yanagida, Y., & Aizawa, M. (2000) · Analytical Biochemistry · doi:10.1006/abio.2000.4584 · Cited by 19 · see citing works
Tang, J., Yang, H., Liang, S., Chen, Y., Mu, Q., & Zhang, J. (2012) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-012-4303-x · Cited by 8 · see citing works
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Secretion of recombinant proteins from E. coli
Kleiner‐Grote, G. R. M., Risse, J. M., & Friehs, K. (2018) · Engineering in Life Sciences · doi:10.1002/elsc.201700200 · Cited by 159 · 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
Zhou, Y., Lu, Z., Wang, X., Selvaraj, J. N., & Zhang, G. (2018) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-017-8700-z · Cited by 97 · see citing works
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Chen, Y., Wu, C., Shen, Y., Ma, Y., Wei, D., & Wang, W. (2019) · Biotechnology for Biofuels · doi:10.1186/s13068-019-1375-1 · Cited by 86 · see citing works
Evolving Paradigms of Recombinant Protein Production in Pharmaceutical Industry: A Rigorous Review
Jayakrishnan, A., Rosli, W. R. W., Tahir, A. R. M., Razak, F. S. A., Kee, P. E., Ng, H. S., Chew, Y.-L., Lee, S.-K., Ramasamy, M., Tan, C. S., et al. (2024) · Sci · doi:10.3390/sci6010009 · Cited by 65 · see citing works
Boosting Secretion of Extracellular Protein by Escherichia coli via Cell Wall Perturbation
Yang, H., Lu, X., Hu, J., Chen, Y., Shen, W., & Liu, L. (2018) · Applied and Environmental Microbiology · doi:10.1128/aem.01382-18 · Cited by 47 · see citing works
Self-citationDegradation of Aflatoxin B1 by recombinant laccase extracellular produced from Escherichia coli
Bian, L., Zheng, M., Chang, T., Zhou, J., & Zhang, C. (2022) · Ecotoxicology and Environmental Safety · doi:10.1016/j.ecoenv.2022.114062 · Cited by 45 · see citing works
The Rocky Road From Fed-Batch to Continuous Processing With E. coli
Kopp, J., Slouka, C., Spadiut, O., & Herwig, C. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00328 · Cited by 42 · see citing works
Bioprocessing of inclusion bodies from E. coli. to produce bioactive recombinant proteins
Rani, A. K., Katiyar, R., & Rathore, A. S. (2023) · Biochemical Engineering Journal · doi:10.1016/j.bej.2023.109188 · Cited by 24 · see citing works
Monitoring and control of E. coli cell integrity
Kastenhofer, J., Rajamanickam, V., Libiseller-Egger, J., & Oliver Spadiut, A. (2021) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2021.01.009 · Cited by 22 · see citing works
Pang, C., Liu, S., Zhang, G., Zhou, J., Du, G., & Li, J. (2022) · Microbial Cell Factories · doi:10.1186/s12934-022-01772-x · Cited by 20 · see citing works
Vitharana, S., Stillahn, J. M., Katayama, D. S., Henry, C. S., & Manning, M. C. (2023) · Journal of Pharmaceutical Sciences · doi:10.1016/j.xphs.2023.08.003 · Cited by 17 · see citing works
Soares, A., Gomes, L. C., Monteiro, G. A., & Mergulhão, F. J. (2021) · Applied Sciences · doi:10.3390/app11188667 · Cited by 14 · see citing works
Ünver, Y., Arslan, Ş. Y., & Acar, M. (2022) · Bioprocess and Biosystems Engineering · doi:10.1007/s00449-021-02678-5 · Cited by 11 · see citing works
Hu, J., Lu, X., Wang, H., Wang, F., Zhao, Y., Shen, W., Yang, H., & Chen, X. (2019) · Engineering in Life Sciences · doi:10.1002/elsc.201800199 · Cited by 11 · see citing works
Self-citationIsmail, A., & Illias, R. M. (2017) · Journal of Industrial Microbiology & Biotechnology · doi:10.1007/s10295-017-1980-6 · Cited by 10 · see citing works
Rezaei, L., Shojaosadati, S. A., Farahmand, L., & Moradi‐Kalbolandi, S. (2020) · Engineering in Life Sciences · doi:10.1002/elsc.201900158 · Cited by 9 · see citing works
He, X., Li, Y., Tao, Y., Qi, X., Ma, R., Jia, H., Yan, M., Chen, K., & Hao, N. (2021) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2021.01.115 · Cited by 9 · see citing works
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He, X., Zhang, S., Dang, D., Lin, T., Ge, Y., Chen, X., & Fan, J. (2023) · Microbial Cell Factories · doi:10.1186/s12934-022-02005-x · Cited by 8 · see citing works
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Hoeven, R., Hughes, J. M. X., Amer, M., Wojcik, E. Z., Tait, S., Faulkner, M., Yunus, I. S., Hardman, S. J. O., Johannissen, L. O., Chen, G., et al. (2019) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/640474 · Cited by 8 · see citing works
Na, K.-I., Kim, S.-J., Choi, D.-S., Min, W.-K., Kim, S.-G., & Seo, J.-H. (2019) · Letters in Applied Microbiology · doi:10.1111/lam.13110 · Cited by 7 · see citing works
Lee, H., Kang, T., Kim, Y.-W., Lee, H., & Kim, S. (2021) · Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2021.109918 · Cited by 6 · see citing works
Kang, T., Hong, S.-H., Jeon, G., Yang, Y., & Kim, S. (2021) · Journal of Industrial Microbiology & Biotechnology · doi:10.1093/jimb/kuab032 · Cited by 4 · see citing works
Lu, J., & Zhang, J. (2019) · RSC Advances · doi:10.1039/c9ra04765d · Cited by 4 · see citing works
Li, J., Wu, B., Ji, Y., Zhang, S., Ge, Y., & Fan, J. (2024) · Protein Expression and Purification · doi:10.1016/j.pep.2024.106617 · Cited by 2 · see citing works
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Coşkun, K. A., Yurekli, N., Abay, E. C., Tutar, M., Al, M., & Tutar, Y. (2022) · Biochemistry · doi:10.5772/intechopen.103958 · Cited by 2 · see citing works
Matias, S. C. B., Azevedo, B., Filho, J. D. B. D. C., Lima, M. Y. L., Moura, A. D., Martins, D. R. A., Júnior, F. C. D. S., & dos Santos, E. S. (2022) · Protein Expression and Purification · doi:10.1016/j.pep.2022.106173 · Cited by 2 · see citing works
Nik-Pa, N. I. M., Abd‐Aziz, S., Ibrahim, M. F., Alitheen, N. B., & Ramli, N. (2019) · Asia-Pacific Journal of Molecular Biology and Biotechnology · doi:10.35118/apjmbb.2019.027.2.12 · Cited by 1 · see citing works
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Chen, X., Li, J., Ji, Y., Zhang, S., Xie, Y., Ge, Y., Wu, B., & Fan, J. (2024) · Research Square · doi:10.21203/rs.3.rs-3981174/v1
Pang, C., Liu, S., Zhang, G., Zhou, J., Du, G., & Li, J. (2022) · Figshare · doi:10.6084/m9.figshare.19387780
- Authors
- Bao, R.-M., Yang, H.-M., Yu, C.-M., Zhang, W.-F., & Tang, J.-B.
- Affiliations
- Weifang Medical University (CN)
- Venue
- Protein Expression and Purification
- Publisher
- Elsevier BV
- Published
- 2016-10
- Volume
- 126
- Pages
- 9-15
- ISSN
- 1046-5928 (ISSN-L)
- OpenAlex
- W2358813103
- Semantic Scholar
- 9abb69ee4b0f3e436a5d8d217a65cff338762e20
- Cited by
- 31
- References
- 41
- Citations by year
- 2024: 3 · 2023: 3 · 2022: 6 · 2021: 5 · 2020: 3 · 2019: 6 · 2018: 3 · 2017: 2
- FWCI
- 2.46
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Protein Expression and Purification (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Natural Scientific Foundation of China (81201346, 81573717)
- Natural Scientific Foundation of Shandong Province (ZR2013HL066, ZR2012CM025)
- Ministry of Science and Technology (2013GA740103)
- Weifang Medical University (K11TS1001)
- 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
- extracellular protein secretion, recombinant protein expression, Escherichia coli, sucrose, glycine, Triton X-100, alkaline phosphatase, IgG-binding protein, cell wall integrity, culture medium optimization, immunoassay reagents
- MeSH terms
- Alkaline Phosphatase, Escherichia coli, Glycine, Recombinant Fusion Proteins, Sucrose, Gene Expression, Octoxynol, Periplasm
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