Abstract
Escherichia coli is the most widely used heterologous protein expression system. However, this system remains a challenge due to the low solubility of proteins, insufficient yield, and inclusion body formation. Numerous approaches have sought to address these issues. The use of a fusion tag is one of the most powerful strategies for obtaining large amounts of heterologous protein in E. coli expression system. Here, recent advances in fusion tags that increase the expression of proteins are reviewed. In addition, proposed concepts for designing peptide tags to increase protein expression are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Achmuller C, Kaar W, Ahrer K, Wechner P, Hahn R, Werther F, Schmidinger H, Cserjan-Puschmann M, Clementschitsch F, Striedner G, Bayer K, Jungbauer A, Auer B (2007) N(pro) fusion technology to produce proteins with authentic N termini in E. coli. Nat Methods 4:1037–1043. https://doi.org/10.1038/nmeth1116
Ahmad T, Singh RS, Gupta G, Sharma A, Kaur B (2019) Chapter 15 Metagenomics in the search for industrial enzymes. Adv Enzyme Technol:419–451. https://doi.org/10.1016/b978-0-444-64114-4.00015-7
Arena TA, Chou B, Harms PD, Wong AW (2019) An anti-apoptotic HEK293 cell line provides a robust and high titer platform for transient protein expression in bioreactors. mAbs 11:977–986. https://doi.org/10.1080/19420862.2019.1598230
Assumpcao MC, de Souza EM, Yates MG, de Oliveira PF, Benelli EM (2007) Purification and characterisation of Azospirillum brasilense N-truncated NtrX protein. Protein Expr Purif 53:302–308. https://doi.org/10.1016/j.pep.2007.01.003
Baeshen NA, Baeshen MN, Sheikh A, Bora RS, Ahmed MMM, Ramadan HAI, Saini KS, Redwan EM (2014) Cell factories for insulin production. Microb Cell Factories 13:141. https://doi.org/10.1186/s12934-014-0141-0
Bañuelos-Hernández B (2017) Algevir: an expression system for microalgae based on viral vectors. Front Microbiol 8:1100. https://doi.org/10.3389/fmicb.2017.01100
Bernier SC, Cantin L, Salesse C (2018) Systematic analysis of the expression, solubility and purification of a passenger protein in fusion with different tags. Protein Expr Purif 152:92–106. https://doi.org/10.1016/j.pep.2018.07.007
Bogomolovas J, Simon B, Sattler M, Stier G (2009) Screening of fusion partners for high yield expression and purification of bioactive viscotoxins. Protein Expr Purif 64:16–23. https://doi.org/10.1016/j.pep.2008.10.003
Butt TR, Edavettal SC, Hall JP, Mattern MR (2005) SUMO fusion technology for difficult-to-express proteins. Protein Expr Purif 43:1–9. https://doi.org/10.1016/j.pep.2005.03.016
Byrne LJ, Sidhu A, Wallis AK, Ruddock LW, Freedman RB, Howard MJ, Williamson RA (2009) Mapping of the ligand-binding site on the b' domain of human PDI: interaction with peptide ligands and the x-linker region. Biochem J 423:209–217. https://doi.org/10.1042/BJ20090565
Caroccia KE, Estephan R, Cohen LS, Arshava B, Hauser M, Zerbe O, Becker JM, Naider F (2011) Expression and biophysical analysis of a triple-transmembrane domain-containing fragment from a yeast G protein-coupled receptor. Biopolymers 96:757–771. https://doi.org/10.1002/bip.21614
Carson M, Johnson DH, McDonald H, Brouillette C, Delucas LJ (2007) His-tag impact on structure. Acta Crystall Sec D, Biol Crystallogr 63:295–301. https://doi.org/10.1107/S0907444906052024
Cereghino JL, Cregg JM (2000) Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev 24:45–66. https://doi.org/10.1111/j.1574-6976.2000.tb00532.x
Chen X, Shi J, Chen R, Wen Y, Shi Y, Zhu Z, Guo S, Li L (2015) Molecular chaperones (TrxA, SUMO, Intein, and GST) mediating expression, purification, and antimicrobial activity assays of plectasin in Escherichia coli. Biotechnol Appl Biochem 62:606–614. https://doi.org/10.1002/bab.1303
Chen K-J, Wu Y-T, Lee C-K (2019) Cellulose binding domain fusion enhanced soluble expression of fructosyl peptide oxidase and its simultaneous purification and immobilization. Int J Biol Macromol 133:980–986. https://doi.org/10.1016/j.ijbiomac
Cheng C, Wu S, Cui L, Wu Y, Jiang T, He B (2017) A novel Ffu fusion system for secretory expression of heterologous proteins in Escherichia coli. Microb Cell Factories 16:231. https://doi.org/10.1186/s12934-017-0845-z
Chu J, Wu X, Wu B, Wang R, He B (2014) Characteristics of an organic solvent-tolerant β-fructofuranosidase from Arthrobacter arilaitensis NJEM01 and efficient synthesis of prebiotic kestose. J Agric Food Chem 62:5408–5411. https://doi.org/10.1021/jf5020523
Costa SJ, Almeida A, Castro A, Domingues L, Besir H (2012) The novel Fh8 and H fusion partners for soluble protein expression in Escherichia coli: a comparison with the traditional gene fusion technology. Appl Microbiol Biotechnol 97:6779–6791. https://doi.org/10.1007/s00253-012-4559-1
Costa SJ, Almeida A, Castro A, Domingues L, Besir H (2013) The novel Fh8 and H fusion partners for soluble protein expression in Escherichia coli: a comparison with the traditional gene fusion technology. Appl Microbiol Biotechnol 97:6779–6791. https://doi.org/10.1007/s00253-012-4559-1
Costa S, Almeida A, Castro A, Domingues L (2014) Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system. Front Microbiol 5:63. https://doi.org/10.3389/fmicb.2014.00063
de Marco A (2009) Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli. Microb Cell Factories 8:26. https://doi.org/10.1186/1475-2859-8-26
Do BH, Ryu HB, Hoang P, Koo BK, Choe H (2014) Soluble prokaryotic overexpression and purification of bioactive human granulocyte colony-stimulating factor by maltose binding protein and protein disulfide isomerase. PLoS One 9:e89906. https://doi.org/10.1371/journal.pone.0089906
Douette P, Navet R, Gerkens P, Galleni M, Lévy D, Sluse FE (2005) Escherichia coli fusion carrier proteins act as solubilizing agents for recombinant uncoupling protein 1 through interactions with GroEL. Biochem Biophys Res Commun 333:686–693. https://doi.org/10.1016/j.bbrc.2005.05.164
Fahad S, Khan FA, Pandupuspitasari NS, Ahmed MM, Liao YC, Waheed MT, Sameeullah M, Darkhshan HS, Saud S, Hassan S, Jan A, Jan MT, Wu C, Chun MX, Huang J (2015) Recent developments in therapeutic protein expression technologies in plants. Biotechnol Lett 37:265–279. https://doi.org/10.1007/s10529-014-1699-7
Fossum S, Crooke E, Skarstad K (2007) Organization of sister origins and replisomes during multifork DNA replication in Escherichia coli. EMBO J 26:4514–4522. https://doi.org/10.1038/sj.emboj.7601871
Furukawa H, Haga T (2000) Expression of functional M2 muscarinic acetylcholine receptor in Escherichia coli. J Biochem 127:151–161. https://doi.org/10.1093/oxfordjournals.jbchem.a022577
Galligan JJ, Petersen DR (2012) The human protein disulfide isomerase gene family. Hum Genom 6:6. https://doi.org/10.1186/1479-7364-6-6
Gangl D, Zedler JAZ, Rajakumar PD, Martinez EMR, Riseley A, Włodarczyk A, Purton S, Sakuragi Y, Howe CJ, Jensen PE, Robinson C (2015) Biotechnological exploitation of microalgae. J Exp Bot 66:6975–6990. https://doi.org/10.1093/jxb/erv426
García-Fruitós E, González-Montalbán N, Morell M, Vera A, Ferraz R, Arís A, Ventura S, Villaverde A (2005) Aggregation as bacterial inclusion bodies does not imply inactivation of enzymes and fluorescent proteins. Microb Cell Factories 4:27. https://doi.org/10.1186/1475-2859-4-27
Gaudry A, Lorber B, Neuenfeldt A, Sauter C, Florentz C, Sissler M (2012) Re-designed N-terminus enhances expression, solubility and crystallizability of mitochondrial protein. Prot Eng Des Sel 25:473–481. https://doi.org/10.1093/protein/gzs046
Geisse S, Fux C (2009) Chapter 15 Recombinant protein production by transient gene transfer into mammalian cells. Methods Enzymol 463:223–238. https://doi.org/10.1016/s0076-6879(09)63015-9
Goda N, Matsuo N, Tenno T, Ishino S, Ishino Y, Fukuchi S, Ota M, Hiroaki H (2015) An optimized Npro-based method for the expression and purification of intrinsically disordered proteins for an NMR study. Intrinsic Disord Proteins 3:e1011004. https://doi.org/10.1080/21690707.2015.1011004
Graether SP (2019) Troubleshooting guide to expressing intrinsically disordered proteins for use in NMR experiments. Front Mol Biosci 5:118. https://doi.org/10.3389/fmolb.2018.00118
Grunina TM, Demidenko AV, Lyaschuk AM, Poponova MS, Galushkina ZM, Soboleva LA, Cherepushkin SA, Polyakov NB, Grumov DA, Solovyev AI, Zhukhovitsky VG, Boksha IS, Subbotina ME, Gromov AV, Lunin VG, Karyagina AS (2017) Recombinant human erythropoietin with additional processable protein domains: purification of protein synthesized in Escherichia coli heterologous expression system. Biochemistry (Mosc) 82:1285–1294. https://doi.org/10.1134/S0006297917110062
Guo Y, Yu M, Jing N, Zhang S (2018) Production of soluble bioactive mouse leukemia inhibitory factor from Escherichia coli using MBP tag. Protein Expr Purif 150:86–91. https://doi.org/10.1016/j.pep.2018.05.006
Haberstock S, Roos C, Hoevels Y, Dötsch V, Schnapp G, Pautsch A, Bernhard F (2012) A systematic approach to increase the efficiency of membrane protein production in cell-free expression systems. Protein Expr Purif 82:308–316. https://doi.org/10.1016/j.pep.2012.01.018
Hage N, Renshaw JG, Winkler GS, Gellert P, Stolnik S, Falcone FH (2015) Improved expression and purification of the Helicobacter pylori adhesin BabA through the incorporation of a hexa-lysine tag. Protein Expr Purif 106:25–30. https://doi.org/10.1016/j.pep.2014.10.009
Han Y, Guo W, Su B, Guo Y, Wang J, Chu B, Yang G (2018) High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag. Protein Expr Purif 142:25–31. https://doi.org/10.1016/j.pep.2017.09.013
Harper S, Speicher DW (2011) Purification of proteins fused to glutathione S-transferase. Methods Mol Biol 681:259–280. https://doi.org/10.1007/978-1-60761-913-0_14
Hirose S, Kawamura Y, Mori M, Yokota K, Noguchi T, Goshima N (2011) Development and evaluation of data-driven designed tags (DDTs) for controlling protein solubility. New Biotechnol 28:225–231. https://doi.org/10.1016/j.nbt.2010.08.012
Hwang PM, Pan JS, Sykes BD (2012) A PagP fusion protein system for the expression of intrinsically disordered proteins in Escherichia coli. Protein Expr Purif 85:148–151. https://doi.org/10.1016/j.pep.2012.07.007
Hwang PM, Pan JS, Sykes BD (2014) Targeted expression, purification, and cleavage of fusion proteins from inclusion bodies in Escherichia coli. FEBS Lett 588:247–252. https://doi.org/10.1016/j.febslet.2013.09.028
Isalan M, Chang KY, Yang J-R (2013) Analysis and prediction of highly effective antiviral peptides based on random forests. PLoS One 8:e70166. https://doi.org/10.1371/journal.pone.0070166
Islam MM, Nakamura S, Noguchi K, Yohda M, Kidokoro S-i, Kuroda Y (2015) Analysis and control of protein crystallization using short peptide tags that change solubility without affecting structure, thermal stability, and function. Cryst Growth Des 15:2703–2711. https://doi.org/10.1021/acs.cgd.5b00010
Juturu V, Wu JC (2018) Heterologous protein expression in Pichia pastoris: latest research progress and applications. ChemBioChem 19:7–21. https://doi.org/10.1002/cbic.201700460
Kato A, Maki K, Ebina T, Kuwajima K, Soda K, Kuroda Y (2007) Mutational analysis of protein solubility enhancement using short peptide tags. Biopolymers 85:12–18. https://doi.org/10.1002/bip.20596
Kefala G, Kwiatkowski W, Esquivies L, Maslennikov I, Choe S (2007) Application of Mistic to improving the expression and membrane integration of histidine kinase receptors from Escherichia coli. J Struct Funct Genom 8:167–172. https://doi.org/10.1007/s10969-007-9033-4
Khalilzadeh R, Shojaosadati SA, Maghsoudi N, Mohammadian-Mosaabadi J, Mohammadi MR, Bahrami A, Maleksabet N, Nassiri-Khalilli MA, Ebrahimi M, Naderimanesh H (2004) Process development for production of recombinant human interferon expressed in Escherichia coli. J Ind Microbiol Biotechnol 31:63–69. https://doi.org/10.1007/s10295-004-0117-x
Khan S, Ullah MW, Siddique R, Nabi G, Manan S, Yousaf M, Hou H (2016) Role of recombinant DNA technology to improve life. Int J Genom 2016:1–14. https://doi.org/10.1155/2016/2405954
Ki M-R, Kanth BK, Min KH, Lee J, Pack SP (2012) Increased expression level and catalytic activity of internally-duplicated carbonic anhydrase from Dunaliella species by reconstitution of two separate domains. Process Biochem 47:1423–1427. https://doi.org/10.1016/j.procbio.2012.05.005
Ki M-R, Nguyen TKM, Kim SH, Kwon I, Pack SP (2016) Chimeric protein of internally duplicated α-type carbonic anhydrase from Dunaliella species for improved expression and CO2 sequestration. Process Biochem 51:1222–1229. https://doi.org/10.1016/j.procbio.2016.05.013
Kim JS, Raines RT (1993) Ribonuclease S-peptide as a carrier in fusion proteins. Protein Sci 2:348–356. https://doi.org/10.1002/pro.5560020307
Kim CW, Han KS, Ryu KS, Kim BH, Kim KH, Choi SI, Seong BL (2007) N-terminal domains of native multidomain proteins have the potential to assist de novo folding of their downstream domains in vivo by acting as solubility enhancers. Protein Sci 16:635–643. https://doi.org/10.1110/ps.062330907
Kim S-G, Min W-K, Rho Y-T, Seo J-H (2012) Electrostatic interaction-induced inclusion body formation of glucagon-like peptide-1 fused with ubiquitin and cationic tag. Protein Expr Purif 84:38–46. https://doi.org/10.1016/j.pep.2012.04.010
Kim DS, Kim SW, Song JM, Kim SY, Kwon KC (2019) A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag. BMC Biotechnol 19:13. https://doi.org/10.1186/s12896-019-0506-x
Klocke M, Mundt K, Idler F, Jung S, Backhausen JE (2005) Heterologous expression of enterocin A, a bacteriocin from Enterococcus faecium, fused to a cellulose-binding domain in Escherichia coli results in a functional protein with inhibitory activity against Listeria. Appl Microbiol Biotechnol 67:532–538. https://doi.org/10.1007/s00253-004-1838-5
Knorre WA, Deckwer WD, Korz D, Pohl HD, Riesenberg D, Ross A, Sanders E, Schulz V (1991) High cell density fermentation of recombinant Escherichia coli with computer-controlled optimal growth rate. Ann N Y Acad Sci 646(1 recombinant D):300–306. https://doi.org/10.1111/j.1749-6632.1991.tb18592.x
Knowles TJ, Scott-Tucker A, Overduin M, Henderson IR (2009) Membrane protein architects: the role of the BAM complex in outer membrane protein assembly. Nat Rev Microbiol 7:206–214. https://doi.org/10.1038/nrmicro2069
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132. https://doi.org/10.1016/0022-2836(82)90515-0
Lagassé HAD, Alexaki A, Simhadri VL, Katagiri NH, Jankowski W, Sauna ZE, Kimchi-Sarfaty C (2017) Recent advances in (therapeutic protein) drug development. F1000Res 6:113. https://doi.org/10.12688/f1000research.9970.1
Lawrence MS, Phillips KJ, Liu DR (2007) Supercharging proteins can impart unusual resilience. J Am Chem Soc 129:10110–10112. https://doi.org/10.1021/ja071641y
Le LTM, Nyengaard JR, Golas MM, Sander B (2018) Vectors for expression of signal peptide-dependent proteins in Baculovirus/insect cell systems and their application to expression and purification of the high-affinity immunoglobulin gamma Fc receptor I in complex with its gamma chain. Mol Biotechnol 60:31–40. https://doi.org/10.1007/s12033-017-0041-8
Lee JH, Kim JH, Hwang SW, Lee WJ, Yoon HK, Lee HS, Hong SS (2000) High-level expression of antimicrobial peptide mediated by a fusion partner reinforcing formation of inclusion bodies. Biochem Biophys Res Commun 277:575–580. https://doi.org/10.1006/bbrc.2000.3712
Lee JS, Grav LM, Lewis NE, Faustrup Kildegaard H (2015a) CRISPR/Cas9-mediated genome engineering of CHO cell factories: application and perspectives. Biotechnol J 10:979–994. https://doi.org/10.1002/biot.201500082
Lee KA, Lee S-S, Kim SY, Choi AR, Lee J-H, Jung K-H (2015b) Mistic-fused expression of algal rhodopsins in Escherichia coli and its photochemical properties. Biochim Biophys Acta (BBA) 1850:1694–1703. https://doi.org/10.1016/j.bbagen.2015.04.002
Levy I, Shoseyov O (2002) Cellulose-binding domains. Biotechnol Adv 20:191–213. https://doi.org/10.1016/s0734-9750(02)00006-x
Li Y (2011) Recombinant production of antimicrobial peptides in Escherichia coli: a review. Protein Expr Purif 80:260–267. https://doi.org/10.1016/j.pep.2011.08.001
Li JF, Zhang J, Zhang Z, Kang CT, Zhang SQ (2011) SUMO mediating fusion expression of antimicrobial peptide CM4 from two joined genes in Escherichia coli. Curr Microbiol 62:296–300. https://doi.org/10.1007/s00284-010-9705-3
Lin Z, Zhao Q, Xing L, Zhou B, Wang X (2015) Aggregating tags for column-free protein purification. Biotechnol J 10:1877–1886. https://doi.org/10.1002/biot.201500299
Liu M, Wang B, Wang F, Yang Z, Gao D, Zhang C, Ma L, Yu X (2019) Soluble expression of single-chain variable fragment (scFv) in Escherichia coli using superfolder green fluorescent protein as fusion partner. Appl Microbiol Biotechnol 103:6071–6079. https://doi.org/10.1007/s00253-019-09925-6
Locatelli-Hoops SC, Gorshkova I, Gawrisch K, Yeliseev AA (2013) Expression, surface immobilization, and characterization of functional recombinant cannabinoid receptor CB2. Biochim Biophys Acta (BBA) 1834:2045–2056. https://doi.org/10.1016/j.bbapap.2013.06.003
López-Garrido J, Puerta-Fernández E, Casadesús J (2014) A eukaryotic-like 3′ untranslated region in Salmonella enterica hilD mRNA. Nucleic Acids Res 42:5894–5906. https://doi.org/10.1093/nar/gku222
Lyukmanova EN, Shenkarev ZO, Khabibullina NF, Kulbatskiy DS, Shulepko MA, Petrovskaya LE, Arseniev AS, Dolgikh DA, Kirpichnikov MP (2012) N-terminal fusion tags for effective production of g-protein-coupled receptors in bacterial cell-free systems. Acta Nat 4:58–64
Ma RJ, Wang YH, Liu L, Bai LL, Ban R (2018) Production enhancement of the extracellular lipase LipA in Bacillus subtilis: effects of expression system and Sec pathway components. Protein Expr Purif 142:81–87. https://doi.org/10.1016/j.pep.2017.09.011
Madzak C (2015) Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering. Appl Microbiol Biotechnol 99:4559–4577. https://doi.org/10.1007/s00253-015-6624-z
Malhotra A (2009) Chapter 16 Tagging for protein expression. Methods Enzymol 463:239–258. https://doi.org/10.1016/s0076-6879(09)63016-0
Mamat U, Wilke K, Bramhill D, Schromm AB, Lindner B, Kohl TA, Corchero JL, Villaverde A, Schaffer L, Head SR, Souvignier C, Meredith TC, Woodard RW (2015) Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins. Microb Cell Factories 14:57. https://doi.org/10.1186/s12934-015-0241-5
Marino J, Bordag N, Keller S, Zerbe O (2015) Mistic’s membrane association and its assistance in overexpression of a human GPCR are independent processes. Protein Sci 24:38–48. https://doi.org/10.1002/pro.2582
Mathiesen G, Sveen A, Piard JC, Axelsson L, Eijsink VGH (2008) Heterologous protein secretion by Lactobacillus plantarum using homologous signal peptides. J Appl Microbiol 105:215–226. https://doi.org/10.1111/j.1365-2672.2008.03734.x
McCusker EC, Bane SE, O'Malley MA, Robinson AS (2008) Heterologous GPCR expression: a bottleneck to obtaining crystal structures. Biotechnol Prog 23:540–547. https://doi.org/10.1021/bp060349b
Mo Q, Fu A, Lin Z, Wang W, Gong L, Li W (2018) Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology in Escherichia coli. Lett Appl Microbiol 67:606–613. https://doi.org/10.1111/lam.13079
Murashima K, Kosugi A, Doi RH (2003) Solubilization of cellulosomal cellulases by fusion with cellulose-binding domain of noncellulosomal cellulase engd from Clostridium cellulovorans. Proteins 50:620–628. https://doi.org/10.1002/prot.10298
Mustoe AM, Corley M, Laederach A, Weeks KM (2018) Messenger RNA structure regulates translation initiation: a mechanism exploited from bacteria to humans. Biochemistry 57:3537–3539. https://doi.org/10.1021/acs.biochem.8b00395
Nautiyal K, Kuroda Y (2018) A SEP tag enhances the expression, solubility and yield of recombinant TEV protease without altering its activity. New Biotechnol 42:77–84. https://doi.org/10.1016/j.nbt.2018.02.006
Needle D, Waugh DS (2014) Rescuing aggregation-prone proteins in Escherichia coli with a dual His(6)-MBP tag. Methods Mol Biol 1177:81–94. https://doi.org/10.1007/978-1-4939-1034-2_7
Nevalainen KM, Te'o VS, Bergquist PL (2005) Heterologous protein expression in filamentous fungi. Trends Biotechnol 23:468–474. https://doi.org/10.1016/j.tibtech.2005.06.002
Nguyen MT, Koo BK, Thi Vu TT, Song JA, Chong SH, Jeong B, Ryu HB, Moh SH, Choe H (2014) Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase. PLoS One 9:e89038. https://doi.org/10.1371/journal.pone.0089038
Nguyen MT, Krupa M, Koo BK, Song JA, Vu TT, Do BH, Nguyen AN, Seo T, Yoo J, Jeong B, Jin J, Lee KJ, Oh HB, Choe H (2016) Prokaryotic soluble overexpression and purification of human VEGF165 by fusion to a maltose binding protein tag. PLoS One 11:e0156296. https://doi.org/10.1371/journal.pone.0156296
Nguyen AN, Song J-A, Nguyen MT, Do BH, Kwon GG, Park SS, Yoo J, Jang J, Jin J, Osborn MJ, Jang YJ, Thi Vu TT, Oh H-B, Choe H (2017) Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein. Sci Rep 7:16139. https://doi.org/10.1038/s41598-017-16167-x
Nguyen MT, Prima MJ, Song JA, Kim J, Do BH, Yoo J, Park S, Jang J, Lee S, Lee E, Novais MP, Seo HB, Lee SY, Cho ML, Kim CJ, Jang YJ, Choe H (2019a) Prokaryotic soluble overexpression and purification of oncostatin M using a fusion approach and genetically engineered E. coli strains. Sci Rep 9:13706. https://doi.org/10.1038/s41598-019-50110-6
Nguyen TKM, Ki MR, Son RG, Pack SP (2019b) The NT11, a novel fusion tag for enhancing protein expression in Escherichia coli. Appl Microbiol Biotechnol 103:2205–2216. https://doi.org/10.1007/s00253-018-09595-w
Pandey A, Sarker M, Liu X-Q, Rainey JK (2014) Small expression tags enhance bacterial expression of the first three transmembrane segments of the apelin receptor. Biochem Cell Biol 92:269–278. https://doi.org/10.1139/bcb-2014-0009
Paraskevopoulou V, Falcone F (2018) Polyionic tags as enhancers of protein solubility in recombinant protein expression. Microorganisms 6:47. https://doi.org/10.3390/microorganisms6020047
Park SH, Mrse AA, Nevzorov AA, Mesleh MF, Oblatt-Montal M, Montal M, Opella SJ (2003) Three-dimensional structure of the channel-forming trans-membrane domain of virus protein “u” (Vpu) from HIV-1. J Mol Biol 333:409–424. https://doi.org/10.1016/j.jmb.2003.08.048
Park SY, Park SH, Choi SK (2012) Active inclusion body formation using Paenibacillus polymyxa PoxB as a fusion partner in Escherichia coli. Anal Biochem 426:63–65. https://doi.org/10.1016/j.ab.2012.04.002
Pedelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS (2006) Engineering and characterization of a superfolder green fluorescent protein. Nat Biotechnol 24:79–88. https://doi.org/10.1038/nbt1172
Potetinova Z, Tantry S, Cohen LS, Caroccia KE, Arshava B, Becker JM, Naider F (2012) Large multiple transmembrane domain fragments of a G protein-coupled receptor: biosynthesis, purification, and biophysical studies. Biopolymers 98:485–500. https://doi.org/10.1002/bip.22122
Priyadarshini P, Singh B (2019) Chapter 15 Computational resources and techniques in enzyme research. Adv Enz Technol:453–468. https://doi.org/10.1016/b978-0-444-64114-4.00016-9
Randolph TW (2012) The two faces of His-tag: immune response versus ease of protein purification. Biotechnol J 7:18–19. https://doi.org/10.1002/biot.201100459
Redwan EM, Baeshen NA, Saini KS, Ramadan HAI, Ahmed MMM, Bora RS, Al-Hejin AM, Baeshen MN (2015) Production of biopharmaceuticals in E. coli: current scenario and future perspectives. J Microbiol Biotechnol 25:953–962. https://doi.org/10.4014/jmb.1412.12079
Rinas U, Garcia-Fruitós E, Corchero JL, Vázquez E, Seras-Franzoso J, Villaverde A (2017) Bacterial inclusion bodies: discovering their better half. Trends Biochem Sci 42:726–737. https://doi.org/10.1016/j.tibs.2017.01.005
Rojas-Lopez M, Zorgani MA, Kelley LA, Bailly X, Kajava AV, Henderson IR, Polticelli F, Pizza M, Rosini R, Desvaux M (2018) Identification of the autochaperone domain in the type Va secretion system (T5aSS): prevalent feature of autotransporters with a β-helical passenger. Front Microbiol 8:2607. https://doi.org/10.3389/fmicb.2017.02607
Roosild TP, Greenwald J, Vega M, Castronovo S, Riek R, Choe S (2005) NMR structure of Mistic, a membrane-integrating protein for membrane protein expression. Science 307:1317–1321. https://doi.org/10.1126/science.1106392
Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5:172. https://doi.org/10.3389/fmicb.2014.00172
Sachdev D, Chirgwin JM (2000) Fusions to maltose-binding protein: control of folding and solubility in protein purification. Methods Enzymol 326:312–321. https://doi.org/10.1016/s0076-6879(00)26062-x
Saraswat M, Musante L, Ravidá A, Shortt B, Byrne B, Holthofer H (2013) Preparative purification of recombinant proteins: current status and future trends. Biomed Res Int 2013:1–18. https://doi.org/10.1155/2013/312709
Savitsky P, Bray J, Cooper CDO, Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O (2010) High-throughput production of human proteins for crystallization: the SGC experience. J Struct Biol 172:3–13. https://doi.org/10.1016/j.jsb.2010.06.008
Seo SW, Yang JS, Kim I, Yang J, Min BE, Kim S, Jung GY (2013) Predictive design of mRNA translation initiation region to control prokaryotic translation efficiency. Metab Eng 15:67–74. https://doi.org/10.1016/j.ymben.2012.10.006
Sharma S, Vaid S, Bhat B, Singh S, Bajaj BK (2019) Chapter 17 Thermostable enzymes for industrial biotechnology. Adv Enz Technol. https://doi.org/10.1016/b978-0-444-64114-4.00017-0
Shoseyov O, Shani Z, Levy I (2006) Carbohydrate binding modules: biochemical properties and novel applications. Microbiol Mol Biol Rev 70:283–295. https://doi.org/10.1128/mmbr.00028-05
Singh RS, Singh T, Pandey A (2019) Chapter 1 Microbial enzymes—an overview. Adv Enz Technol. https://doi.org/10.1016/b978-0-444-64114-4.00001-7
Sockolosky JT, Szoka FC (2013) Periplasmic production via the pET expression system of soluble, bioactive human growth hormone. Protein Expr Purif 87:129–135. https://doi.org/10.1016/j.pep.2012.11.002
Song JA, Koo BK, Chong SH, Kwak J, Ryu HB, Nguyen MT, Vu TT, Jeong B, Kim SW, Choe H (2013) Expression and purification of biologically active human FGF2 containing the b'a' domains of human PDI in Escherichia coli. Appl Biochem Biotechnol 170:67–80. https://doi.org/10.1007/s12010-013-0140-3
Song J-W, Woo J-M, Jung GY, Bornscheuer UT, Park J-B (2016) 3′-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli. Sci Rep 6:29406. https://doi.org/10.1038/srep29406
Sørensen HP, Mortensen KK (2005) Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 115:113–128. https://doi.org/10.1016/j.jbiotec.2004.08.004
Sorensen HP, Mortensen KK (2005) Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli. Microb Cell Factories 4:1. https://doi.org/10.1186/1475-2859-4-1
Specht E, Miyake-Stoner S, Mayfield S (2010) Micro-algae come of age as a platform for recombinant protein production. Biotechnol Lett 32:1373–1383. https://doi.org/10.1007/s10529-010-0326-5
Sun P, Tropea JE, Waugh DS (2011) Enhancing the solubility of recombinant proteins in Escherichia coli by using hexahistidine-tagged maltose-binding protein as a fusion partner. Methods Mol Biol 705:259–274. https://doi.org/10.1007/978-1-61737-967-3_16
Tolmachev V, Hofström C, Malmberg J, Ahlgren S, Hosseinimehr SJ, Sandström M, Abrahmsén L, Orlova A, Gräslund T (2010) HEHEHE-tagged affibody molecule may be purified by IMAC, is conveniently labeled with [99mTc(CO)3]+, and shows improved biodistribution with reduced hepatic radioactivity accumulation. Bioconjug Chem 21:2013–2022. https://doi.org/10.1021/bc1002357
Uversky VN, Yuan S, Xu J, Ge Y, Yan Z, Du G, Wang N (2013) Prokaryotic ubiquitin-like ThiS fusion enhances the heterologous protein overexpression and aggregation in Escherichia coli. PLoS One 8:e62529. https://doi.org/10.1371/journal.pone.0062529
van Oers MM, Pijlman GP, Vlak JM (2014) Thirty years of baculovirus-insect cell protein expression: from dark horse to mainstream technology. J Gen Virol 96:6–23. https://doi.org/10.1099/vir.0.067108-0
Vavrová Ľ, Muchová K, Barák I (2010) Comparison of different Bacillus subtilis expression systems. Res Microbiol 161:791–797. https://doi.org/10.1016/j.resmic.2010.09.004
Vollmer B, Schooley A, Sachdev R, Eisenhardt N, Schneider AM, Sieverding C, Madlung J, Gerken U, Macek B, Antonin W (2012) Dimerization and direct membrane interaction of Nup53 contribute to nuclear pore complex assembly. EMBO J 31:4072–4084. https://doi.org/10.1038/emboj.2012.256
Wang X, Zhou B, Hu W, Zhao Q, Lin Z (2015) Formation of active inclusion bodies induced by hydrophobic self-assembling peptide GFIL8. Microb Cell Factories 14:88. https://doi.org/10.1186/s12934-015-0270-0
Wei X, Wu R, Zhang L, Ahmad B, Si D, Zhang R (2018) Expression, purification, and characterization of a novel hybrid peptide with potent antibacterial activity. Molecules 23:1491. https://doi.org/10.3390/molecules23061491
Wilkinson DL, Harrison RG (1991) Predicting the solubility of recombinant proteins in Escherichia coli. Nat Biotechnol 9:443–448. https://doi.org/10.1038/nbt0591-443
Wright PE, Dyson HJ (2015) Intrinsically disordered proteins in cellular signaling and regulation. Nat Rev Mol Cell Biol 16:18–29. https://doi.org/10.1038/nrm3920
Wu W, Xing L, Zhou B, Lin Z (2011) Active protein aggregates induced by terminally attached self-assembling peptide ELK16 in Escherichia coli. Microb Cell Factories 10:9. https://doi.org/10.1186/1475-2859-10-9
Xing L, Wu W, Zhou B, Lin Z (2011) Streamlined protein expression and purification using cleavable self-aggregating tags. Microb Cell Factories 10:42. https://doi.org/10.1186/1475-2859-10-42
Xu Y, Foong FC (2008) Characterization of a cellulose binding domain from Clostridium cellulovorans endoglucanase-xylanase D and its use as a fusion partner for soluble protein expression in Escherichia coli. J Biotechnol 135:319–325. https://doi.org/10.1016/j.jbiotec.2008.05.004
Yadav DK, Yadav N, Yadav S, Haque S, Tuteja N (2016) An insight into fusion technology aiding efficient recombinant protein production for functional proteomics. Arch Biochem Biophys 612:57–77. https://doi.org/10.1016/j.abb.2016.10.012
Young CL, Britton ZT, Robinson AS (2012) Recombinant protein expression and purification: a comprehensive review of affinity tags and microbial applications. Biotechnol J 7:620–634. https://doi.org/10.1002/biot.201100155
Yuan S, Wang X, Xu J, Yan Z, Wang N (2014) Ubiquitin-like prokaryotic MoaD as a fusion tag for expression of heterologous proteins in Escherichia coli. BMC Biotechnol 14:5. https://doi.org/10.1186/1472-6750-14-5
Zhang Y-B, Howitt J, McCorkle S, Lawrence P, Springer K, Freimuth P (2004) Protein aggregation during overexpression limited by peptide extensions with large net negative charge. Protein Expr Purif 36:207–216. https://doi.org/10.1016/j.pep.2004.04.020
Zhang Z, Tang R, Zhu D, Wang W, Yi L, Ma L (2017) Non-peptide guided auto-secretion of recombinant proteins by super-folder green fluorescent protein in Escherichia coli. Sci Rep 7:6990. https://doi.org/10.1038/s41598-017-07421-3
Zhao X, Li G, Liang S (2013) Several affinity tags commonly used in chromatographic purification. J Anal Methods Chem 2013:581093. https://doi.org/10.1155/2013/581093
Zhao W, Liu L, Du G, Liu S (2018) A multifunctional tag with the ability to benefit the expression, purification, thermostability and activity of recombinant proteins. J Biotechnol 283:1–10. https://doi.org/10.1016/j.jbiotec.2018.07.005
Zhao W, Liu S, Du G, Zhou J (2019) An efficient expression tag library based on self-assembling amphipathic peptides. Microb Cell Factories 18:91. https://doi.org/10.1186/s12934-019-1142-9
Zhou B, Xing L, Wu W, Zhang X-E, Lin Z (2012) Small surfactant-like peptides can drive soluble proteins into active aggregates. Microb Cell Factories 11:10. https://doi.org/10.1186/1475-2859-11-10
Zorko M, Jerala R (2010) Production of recombinant antimicrobial peptides in bacteria. Methods Mol Biol 618:61–76. https://doi.org/10.1007/978-1-60761-594-1_5
Funding
This work was supported by the Marine Biomaterials Research Center as a grant from the Marine Biotechnology Program, funded by the Ministry of Oceans and Fisheries, South Korea. This work is also supported by the Basic Core Technology Development Program for the Oceans and the Polar Regions of the NRF (NRF-2015M1A5A1037054) and the Research Fellow Funding Grant of NRF (NRF-2018R1A6A3A11040793), and supported by a Korea University Grant for MR Ki.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Ki, MR., Pack, S.P. Fusion tags to enhance heterologous protein expression. Appl Microbiol Biotechnol 104, 2411–2425 (2020). https://doi.org/10.1007/s00253-020-10402-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-020-10402-8