Abstract
Leishmaniosis is caused by the protozoa of the genus Leishmania with a wide spectrum of clinical and epidemiological manifestations which are characterized into four clinical groups: cutaneous, mucocutaneous, diffuse cutaneous, and visceral. American visceral leishmaniosis (AVL) or visceral leishmaniosis (VL) has been known as the most severe form of the disease. However, despite the growing number of people exposed to the infection risk and the great effort done by the scientific community worldwide to significantly increase the knowledge about these diseases, there is no vaccine capable of preventing VL in humans. In this short review, we present some of the plasmids used for the expression of recombinant protein by Escherichia coli strains used mainly for the second generation of vaccines for leishmaniosis. It can be emphasized that currently, these vectors and hosts play an important role in developing vaccine strategies against the disease. Indeed, use of the E. coli BL21 (DE) strain is remarkable mainly due to its characteristics for being a stable protein producer as well as the use of histidine tags for antigen purification.
Key points
• Plasmid vectors and E. coli will continue being important for studies about leishmaniosis.
• Protein purification exploiting histidine tags is a key technique.
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References
Ahmad I, Nawaz N, Darwesh NM, Ur Rahman S, Mustafa MZ, Khan SB, Patching SG (2018) Overcoming challenges for amplified expression of recombinant proteins using Escherichia coli. Protein Expr Purif 144:12–18. https://doi.org/10.1016/j.pep.2017.11.005
Ali D, Abbady A-Q, Kweider M, Soukkarieh C (2016) Cloning, expression, purification and characterization of Leishmania tropica PDI-2. Open Life Sci 11:166–176. https://doi.org/10.1515/biol-2016-0022
Arruda MM, Figueiredo FB, Marcelino AP, Barbosa JR, Werneck GL, Noronha EF, Romero GAS (2016) Sensitivity and specificity of parallel or serial serological testing for detection of canine Leishmania infection. Mem Inst Oswaldo Cruz 111(3):168–173
Aucoin MG, McMurray-Beaulieu V, Poulin F, Boivin EB, Chen J, Ardelean FM, Jolicoeur M (2006) Identifying conditions for inducible protein production in Escherichia coli: combining a fed-batch and multiple induction approach. Microb Cell Factories 5:1–13. https://doi.org/10.1186/1475-2859-5-27
Boarino A, Scalone A, Gradoni L, Ferroglio E, Vitale F, Zanatta R, Giuffrida MG, Rosati S (2005) Development of recombinant chimeric antigen expressing Immunodominant B epitopes of Leishmania infantum for serodiagnosis of visceral Leishmaniasis. Clin Diagn Lab Immunol 12(5):647–653
Briand L, Marcion G, Kriznik A, Heydel JM, Artur Y, Garrido C, Seigneuric R, Neiers F (2016) A self-inducible heterologous protein expression system in Escherichia coli. Sci Rep 6(33037):1–11. https://doi.org/10.1038/srep33037
Chaukan K, Kumar G, Kaur S (2018) Activity of rutin, a potent flavonoid against SSG-sensitive and -resistant Leishmania dovani parasites in experimental leishmaniasis. Int Immunopharmacol 64:372–385. https://doi.org/10.1016/j.itimp.2018.09.026
Cheng S, Song Q, Wei D, Gao B (2007) High-level production penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli with optimization of carbon sources. Enzyme Microb Tech 41(3):326–330. https://doi.org/10.1016/j.enzmictec.2007.02.011
Collins T, Azevedo-silva J, Costa A, Branca F, Machado R, Casal M (2013) Batch production of a silk-elastin-like protein in Escherichia coli BL21 ( DE3 ): key parameters for optimisation. Microb Cell Factories 12(21):1–16. https://doi.org/10.1186/1475-2859-12-21
Einsfeldt K, Severo Júnior JB, Corrêa Argondizzo AP, Medeiros MA, Alves TLM, Almeida RV, Larentis AL (2011) Cloning and expression of protease ClpP from Streptococcus pneumoniae in Escherichia coli: study of the influence of kanamycin and IPTG concentration on cell growth, recombinant protein production and plasmid stability. Vaccine 29(41):7136–7143. https://doi.org/10.1016/j.vaccine.2011.05.073
Esteva L, Vargas C, Vargas de León C (2017) The role of asymptomatics and dogs on leishmaniasis propagation. Math Biosci 2:46–55. https://doi.org/10.1016/j.mbs.2017.08.006
Farajnia S, Rhabarnia L, Zanjani BM, Alimohammadian MH, Oskoee SA, Beh-pajooh A, Saeedi N, Saheb SM (2011) Molecular cloning and characterization of P4 nuclease from Leishmania infantum. Enzyme Res 2001:6. https://doi.org/10.4061/2011/970983
Freitas-Junior LH, Chatelain E, Kim HA, Siqueira-Neto JL (2012) Visceral leishmaniasis treatment: what do we have, what do we need and how to deliver it? Int J Parasitol Drugs Drug Resist 2:11–19. https://doi.org/10.1016/j.ijpddr.2012.01.003
Ghorbani M, Farhoudi R (2018) Leishmaniasis in humans: drug or vaccine therapy? Drug Des Devel Ther 12:25–40. https://doi.org/10.2147/DDDT.2012.S146521
Ghorbani D, Sahni G, Sahoo DK (2009) Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture. Bioresour Technol 100(19):4468–4474. https://doi.org/10.1016/j.biortech.2009.04.008
Gillespie PM, Beaumier CM, Strych U, Hayward T, Hotez PJ, Bottazzi ME (2016) Status of vaccine research and development of vaccines for leishmaniasis. Vaccine 34(26):2992–2995. https://doi.org/10.1016/j.vaccine.2015.12.071
Gontijo CMF, Melo MN (2004) Leishmania Viceral no Brasil: Quadro atual desafios e pespectivas. Rev Bras Epidemiol 7(3):338–349
Hammoudeh N, Soukkarieh C, Abbady A-Q, Kweider M (2015) Cloning, expression and purification of recombinant Leishmania tropica P36 (Lack) protein for serodiagnosis and vaccine applications. Int J Pharm Tech Res 7(1):72–79
Jain K, Jain NK (2015) Vaccines for visceral leishmaniasis: a review. J Immunol Methods 422:1–12. https://doi.org/10.1016/j.jim.2015.03.017
Jhamb K, Sahoo DK (2012) Production of soluble recombinant proteins in Escherichia coli: effects of process conditions and chaperone co-expression on cell growth and production of xylanase. Bioresour Technol 123:135–143. https://doi.org/10.1016/j.biortech.2012.07.011
Jusi MMG (2015) Resposta Imune de camundongos BALB/c imunizados com a proteína recombinante A2 de Leishmania chagasi. f 125, Thesis, Universidade Estadual Paulista (UNESP)
Jusi MMG, Oliveira TMFS, Nakaghi ACH, André MR, Machado RZ (2015) Expression of a recombinant protein, A2 family, from Leishmania infantum (Jaboticabal strain) and its evaluation in canine visceral Leishmaniasis serological test. Braz J Vet Parasit 24(3):309–316. https://doi.org/10.1590/S1984-29612015060
Karami M, Doudi M, Setorki M (2013) Assessing epidemiology of cutaneous leishmaniasis in Isfahan, Iran. J Vector Dis 50(1):30–37
Kaur J, Kumar A, Kaur J (2018) Strategies for optimization of heterologous protein expression in Escherichia coli: roadblocks and reinforcements. Int J Biol Macromol 106:803–822. https://doi.org/10.1016/j.ijbiomac.2017.08.080
Larentis AL, Nicolau JFMQ, Esteves GDS, Vareschini DT, de Almeida FVR, dos Reis MG, Medeiros MA (2014) Evaluation of pre-induction temperature, cell growth at induction and IPTG concentration on the expression of a leptospiral protein in Escherichia coli using shaking flasks and microbioreactor. BMC Res Notes 7:671. https://doi.org/10.1186/1756-0500-7-671
Lecina M, Sarró E, Casablancas A, Gòdia F, Cairó JJ (2013) IPTG limitation avoids metabolic burden and acetic acid accumulation in induced fed-batch cultures of Escherichia coli M15 under glucose limiting conditions. Biochem Eng J 70:78–83. https://doi.org/10.1016/j.bej.2012.10.006
Leitão ALOS, Caldas MCB, Padilha CEA, Costa CN, Rocha PM, Sousa Júnior FC, Macedo GR, Santos ES (2018) Recovery and purification of 503 antigen from Leishmania i. chagasi with simultaneous removal of lipopolysaccharides: influence of immobilized metals and elution strategies during expanded bed adsorption (EBA). J Liq Chrom Rel Technol 41(19–20):1066–1073. https://doi.org/10.1080/10826076.2019.1565829
Lima C, Mesquita JR, Brancal H, Vahlenkamp T, Teixeira AR, Cardoso L, Amorim C, Santarém N, Silva AC (2017) The use of Escherichia coli total antigens as a complementary approach to address seropositivity to Leishmania antigens in canine leishmaniosis. Parasitology 144:1384–1393. https://doi.org/10.1017/S0031182017000713
Malik A, Alsenaidy AM, Elrobh M, Khan W, Alanazi MS, Bazzi MD (2016) Optimization of expression and purification of HSPA6 protein from Camelus dromedarius in Escherichia coli. Saudi J Biol Sci 23(3):410–419. https://doi.org/10.1016/j.sjbs.2015.04.017
Marbach A, Bettenbrock K (2012) Lac operon induction in Escherichia coli: systematic comparison of IPTG and TMG induction and influence of the transacetylase LacA. J Biotechnol 157(1):82–88. https://doi.org/10.1016/j.jbiotec.2011.10.009
Martins DRA, Jeronimo SMB, Donelson JE, Wilson ME (2006) Leishmania chagasi T-cell antigens identified through a double library screen. Infect Immun 74(12):6940–6948. https://doi.org/10.1128/IAI.02032-05
Michalsky EM, Rocha MF, Rocha-Lima ACVM, França-Silva JC, Pires MQ, Oliveira FS, Pacheco RS, Santos SL, Barata RA, Romanha AJ, Fortes-Dias CL, Dias ES (2007) Infectivity of seropositive dogs, showing different clinical forms of leishmaniasis, to Lutzomyia longipalpis pheblotomine sand flies. Vet Parasitol 147(1–2):67–76. https://doi.org/10.1016/j.vetpar.2007.03.004
Nogueira CT (2014) Busca de antígenos para diagnóstico da leishmaniose visceral canica: potencial uso e aplicação da proteína rLc36. f 104, Thesis, UNESP/Araragura
Nogueira CT, Cistis MLD, Urbaczek AC, Jusi MMG, Velásque AMA, Machado RZ, Ferreira H, Henrique-Silva F, Langoni H, Costa PI, Graminha MAS (2018) Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis. Mem Inst Oswaldo Cruz 113(3):197–201
Norsyahida A, Rahmah N, Ahmad RMY (2009) Effects of feeding and induction strategy on the production of BmR1 antigen in recombinant Escherichia coli. Lett Appl Microbiol 49(5):544–550. https://doi.org/10.1111/j.1472-765X.2009.02694.x
Porrozi R, Costa MVS, Teva A, Falqueto A, Ferreira AL, Santos CD, Fernandes AP, Gazzinelli RT, Campos-Neto A, Grimaldi G Jr (2007) Comparative evaluation of enzyme-linked immunosorbent assays based on crude and recombinant Leishmanial antigens for serodiagnosis of symptomatic and asymptomatic Leishmania infantum visceral infections in dogs. Clin Vacc Immunol 14(5):544–548
Ramos I, Alonso A, Marcen JM, Peris A, Castillo JA, Colmenares M, Larraga V (2008) Heterologous prime-boost vaccination with a non-replicative vaccinia recombinant vector expressing LACK confers protection against canine visceral leishmaniasis with a predominant Th1-specific immune response. Vaccine 26:333–344. https://doi.org/10.1016/j.vaccine.2007.11.021
Ramos I, Alonso A, Peris A, Marcen JM, Abengozar MA, Alcolea PJ, Castillo JA, Larraga V (2009) Antibiotic resistance free plasmid DNA expressing LACK protein leads towards a protective Th1 response against Leishmania infantum infection. Vaccine 27:6695–6703. https://doi.org/10.1016/j.vaccine.2009.08.091
Ribeiro VT (2018) Avaliação da estabilidade do plasmídeo pQE-30 e expressão do antígeno 503 de Leishmania i. chagasi em Escherichia coli M15 em diferentes concentrações de IPTG e temperaturas de cultivo. f 89, Dissertation, Federal University of Rio Grande do Norte
Ribeiro VT, Asevedo EA, Vasconcelos LTCP, Oliveira Filho MA, Araújo JS, Macedo GR, Sousa Júnior FC, Santos ES (2019) Evaluation of induction conditions for plasmid pQE-30 stability and 503 antigen of Leishmania i. chagasi expression in E. coli M15. Appl Microb Biotechnol 103(16):6495–6504
Rudolph B, Gebendorfer KM, Buchner J, Winter J (2010) Evolution of Escherichia coli for growth at high temperatures. J Biol Chem 285(25):19029–19034. https://doi.org/10.1074/jbc.M110.103374
Selvapandiyan A, Dey R, Gannavaram S, Lakhal-Naouar I, Duncan R, Salotra P, Nakhasi HL (2012) Immunity to visceral leishmaniasis using genetically defined live-attenuated parasites. J Trop Pediatr:1–12. https://doi.org/10.1155/2012/631460
Silva F, Queiroz JA, Domingues FC (2012) Evaluating metabolic stress and plasmid stability in plasmid DNA production by Escherichia coli. Biotechnol Adv 30(3):691–708. https://doi.org/10.1016/j.biotechadv.2011.12.005
Singh OP, Hasker E, Boelaert M, Sundar S (2016) Elimination of visceral leishmaniasis on the Indian subcontinent. Lancet Infect Dis 16(12):304–309. https://doi.org/10.1016/S1473-3099(16)30140-2
Sousa Junior FC, Vaz MRF, Araújo Padilha CE, Chibério AS, Martins DRA, Macedo GR, Santos ES (2015) Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography. J Chromatogr B 986-987:1–7. https://doi.org/10.1016/j.jchromb.2015.01.031
Sousa Junior FC, Ribeiro VT, Chibério AS, Costa LPM, Padilha CEA, Martins DRA, Macedo GR, Santos ES (2017) Simultaneous recombinant 503 antigen recovery and endotoxin removal from E. coli M15 homogenate using expanded bed adsorption chromatography. Separ Sci Technol 52(11):1869–1875. https://doi.org/10.1080/01496395.2017.1305411
Sousa-Gomes ML, Romero GAS, Werneck GL (2017) Visceral leishmaniasis and HIV/AIDS in Brazil: are we aware enough? Plos Neglect Trop D:1–13. https://doi.org/10.1371/journal.pntd.0005772
Tomazetto G, Mulinari F, Stanisçuaski F, Settembrini B, Carlini CR, Ayub MAZ (2007) Expression kinetics and plasmid stability of recombinant Escherichia coli encoding urease-derived peptide with bioinsecticide activity. Enzyme Microb Tech 41(6–7):821–827. https://doi.org/10.1016/j.enzmictec.2007.07.006
Tripathi NK, Sathyaseelan K, Jana AM, Rao PVL (2009) High yield production of heterologous proteins with Escherichia coli. Defence Sci J 59(2):137–146. https://doi.org/10.14429/dsj.59.1501
Ureña-Búrquez LA, Gárcia-Réndon A, Rochín-Wong S, Tejeda-Mansir A (2019) Preparation and characterization of a nanovaccine of pVAX1-NH36 for leishmaniasis. Fuel 235:1077–1082. https://doi.org/10.1016/j.fuel.2018.08.070
Vasconcelos LTCP, Oliveira Filho MA, Ribeiro VT, Araújo JS, Sousa Júnior FC, Martins DRA, Santos ES (2018) Optimization of the 503 antigen induction strategy of Leishmania infantum chagasi expressed in Escherichia coli M15. Prep Biochem Biotechnol 48(10):968–976. https://doi.org/10.1080/10826068.2018.1525563
Vaz MRF, França RLS, Andrade SSL, Sousa Júnior FC, Santos ES, Martins DRA, Macedo GR (2011) Influence of culture medium on the production of EIF antigen from leishmania chagasi in recombinant Escherichia coli. Braz J Microbiol 42(4):1390–1396. https://doi.org/10.1590/S1517-83822011000400021
Vaz MRF, de Sousa Junior FC, Costa LMR, dos Santos ES, Martins DRA, de Macedo GR (2015) Optimization of culture medium for cell growth and expression of 648 antigen from Leishmania infantum chagasi in recombinant Escherichia coli M15. Ann Microbiol 65(3):1607–1613. https://doi.org/10.1007/s13213-014-1000-2
Vélez AM, Silva AD, Horta ACL, Sargo CR, Campani G, Silva GG, Giordano RLC, Zangirolami TC (2014) High-throughput strategies for penicillin G acylase production in rE. coli fed-batch cultivations. BMC Biotechnol 14(1):6. https://doi.org/10.1186/1472-6750-14-6
Wang D, Li Q, Mao Y, Xing J, Su Z (2010) High-level succinic acid production and yield by lactose-induced expression of phosphoenolpyruvate carboxylase in ptsG mutant Escherichia coli. Appl Microbiol Biot 87(6):2025–2035. https://doi.org/10.1007/s00253-010-2689-x
Wen Y, Feng M, Yuan Z, Zhou P (2005) Expression and overproduction of recombinant penicillin G acylase from Kluyvera citrophila in Escherichia coli. Enzyme Microb Tech 37(2):233–237. https://doi.org/10.1016/j.enzmictec.2005.03.005
WHO (2016) World Health Organization. http://www.who.int/mediacentre/factsheets/fs375/%0Aen/.
Zhang H, Zheng Y, Liu Q, Tao X, Zheng W, Ma X, Wei D (2009) Development of a fed-batch process for the production of anticancer drug TATm-survivin (T34A) in Escherichia coli. Biochem Eng J 43(2):163–168. https://doi.org/10.1016/j.bej.2008.09.013
Acknowledgments
The authors acknowledge Dr. Mary Wilson (Iowa University/USA) for providing the microorganism strains at Chemical Engineering Department/UFRN.
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The authors thank Coordination for the Improvement of Higher Education Personnel (CAPES) and Brazilian National Council of Research (CNPq) for the financial support (Grant 401817/2016-4 and 305251/2017-1).
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VTR, ALOSL, LTCPV, MAOF, FCSJ, and ESS conceived and contributed to the literature search. FCSJ, DRAM, and ESS wrote the manuscript. All authors read and approved the manuscript.
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Ribeiro, V.T., de Sá Leitão, A.L.O., de Paiva Vasconcelos, L.T.C. et al. Use of plasmids for expression of proteins from the genus Leishmania in Escherichia coli: current state and perspectives. Appl Microbiol Biotechnol 104, 4273–4280 (2020). https://doi.org/10.1007/s00253-020-10548-5
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DOI: https://doi.org/10.1007/s00253-020-10548-5