Generic placeholder image

Recent Patents on Biotechnology

Editor-in-Chief

ISSN (Print): 1872-2083
ISSN (Online): 2212-4012

Research Article

Evaluating Five Escherichia coli Derivative Strains as a Platform for Arginine Deiminase Overproduction

Author(s): Sara Abdollahi, Mohammad Hossein Morowvat*, Amir Savardashtaki, Cambyz Irajie, Sohrab Najafipour and Younes Ghasemi

Volume 16, Issue 2, 2022

Published on: 11 January, 2022

Page: [174 - 183] Pages: 10

DOI: 10.2174/1872208315666211122114625

Price: $65

Abstract

Aims: This study attempted to evaluate the five host strains, including BL21 (DE3), Rosetta (DE3), DH5α, XL1-BLUE, and SHuffle, in terms of arginine deiminase (ADI) production and enzyme activity.

Background: Escherichia coli is one of the most preferred host microorganisms for the production of recombinant proteins due to its well-characterized genome, availability of various expression vectors, and host strains. Choosing a proper host strain for the overproduction of a desired recombinant protein is very important because of the diversity of genetically modified expression strains. Various E. coli cells have been examined in different patent applications.

Methods: ADI was chosen as a bacterial enzyme that degrades L-arginine. It is effective in the treatment of some types of human cancers like melanoma and hepatocellular carcinoma (HCC), which are arginine-auxotrophic. Five mentioned E. coli strains were cultivated. The pET-3a was used as the expression vector. The competent E. coli cells were obtained through the CaCl2 method. It was then transformed with the construct of pET3a-ADI using the heat shock strategy. The ADI production levels were examined by 10% SDS-PAGE analysis. The ability of host strains for the expression of the requested recombinant protein was compared. The enzymatic activity of the obtained recombinant ADI from each studied strain was assessed by a colorimetric 96-well microtiter plate assay.

Results: All the five strains exhibited a significant band at 46 kDa. BL21 (DE3) produced the highest amount of ADI protein, followed by Rosetta (DE3). The following activity assay showed that ADI from BL21 (DE3) and Rosetta (DE3) had the most activity.

Conclusion: There are some genetic and metabolic differences among the various E. coli strains, leading to differences in the amount of recombinant protein production. The results of this study can be used for the efficacy evaluation of the five studied strains for the production of similar pharmaceutical enzymes. The strains also could be analyzed in terms of proteomics.

Keywords: Arginine deiminase, enzyme activity, Escherichia coli, host cell physiology, recombinant protein production, citrulline.

Graphical Abstract
[1]
Ferrer-Miralles N, Domingo-Espín J, Corchero JL, Vázquez E, Villaverde A. Microbial factories for recombinant pharmaceuticals. Microb Cell Fact 2009; 8: 17.
[http://dx.doi.org/10.1186/1475-2859-8-17] [PMID: 19317892]
[2]
Karyolaimos A, Dolata KM, Antelo-Varela M, et al. Escherichia coli can adapt its protein translocation machinery for enhanced periplasmic recombinant protein production. Front Bioeng Biotechnol 2020; 7: 465.
[http://dx.doi.org/10.3389/fbioe.2019.00465] [PMID: 32064253]
[3]
Rosano GL, Ceccarelli EA. Recombinant protein expression in Escherichia coli: Advances and challenges. Front Microbiol 2014; 5: 172.
[http://dx.doi.org/10.3389/fmicb.2014.00172] [PMID: 24860555]
[4]
Shiloach J, Fass R. Growing E. coli to high cell density- a historical perspective on method development. Biotechnol Adv 2005; 23(5): 345-57.
[http://dx.doi.org/10.1016/j.biotechadv.2005.04.004] [PMID: 15899573]
[5]
Sezonov G, Joseleau-Petit D, D’Ari R. Escherichia coli physiology in Luria-Bertani broth. J Bacteriol 2007; 189(23): 8746-9.
[http://dx.doi.org/10.1128/JB.01368-07] [PMID: 17905994]
[6]
Pope B, Kent HM. High efficiency 5 min transformation of Escherichia coli. Nucleic Acids Res 1996; 24(3): 536-7.
[http://dx.doi.org/10.1093/nar/24.3.536] [PMID: 8602370]
[7]
Sørensen HP, Mortensen KK. Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 2005; 115(2): 113-28.
[http://dx.doi.org/10.1016/j.jbiotec.2004.08.004] [PMID: 15607230]
[8]
Sahdev S, Khattar SK, Saini KS. Production of active eukaryotic proteins through bacterial expression systems: A review of the existing biotechnology strategies. Mol Cell Biochem 2008; 307(1-2): 249-64.
[http://dx.doi.org/10.1007/s11010-007-9603-6] [PMID: 17874175]
[9]
Kang TH, Seong BL. Solubility, stability, and avidity of recombinant antibody fragments expressed in microorganisms. Front Microbiol 2020; 11: 1927.
[http://dx.doi.org/10.3389/fmicb.2020.01927] [PMID: 33101218]
[10]
Sandomenico A, Sivaccumar JP, Ruvo M. Evolution of Escherichia coli expression system in producing a ntibody recombinant fragments. Int J Mol Sci 2020; 21(17): 1-39.
[http://dx.doi.org/10.3390/ijms21176324] [PMID: 32878291]
[11]
Marisch K, Bayer K, Scharl T, et al. A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. PLoS One 2013; 8(8): e70516.
[http://dx.doi.org/10.1371/journal.pone.0070516] [PMID: 23950949]
[12]
Daegelen P, Studier FW, Lenski RE, Cure S, Kim JF. Tracing ancestors and relatives of Escherichia coli B, and the derivation of B strains REL606 and BL21(DE3). J Mol Biol 2009; 394(4): 634-43.
[http://dx.doi.org/10.1016/j.jmb.2009.09.022] [PMID: 19765591]
[13]
Jeong H, Barbe V, Lee CH, et al. Genome sequences of Escherichia coli B strains REL606 and BL21(DE3). J Mol Biol 2009; 394(4): 644-52.
[http://dx.doi.org/10.1016/j.jmb.2009.09.052] [PMID: 19786035]
[14]
Jeong H, Kim HJ, Lee SJ. Complete genome sequence of Escherichia coli strain BL21. Genome Announc 2015; 3(2): e00134-15.
[http://dx.doi.org/10.1128/genomeA.00134-15] [PMID: 25792055]
[15]
Chart H, Smith HR, La Ragione RM, Woodward MJ. An investigation into the pathogenic properties of Escherichia coli strains BLR, BL21, DH5α and EQ1. J Appl Microbiol 2000; 89(6): 1048-58.
[http://dx.doi.org/10.1046/j.1365-2672.2000.01211.x] [PMID: 11123478]
[16]
Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 1986; 189(1): 113-30.
[http://dx.doi.org/10.1016/0022-2836(86)90385-2] [PMID: 3537305]
[17]
Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 1990; 185: 60-89.
[http://dx.doi.org/10.1016/0076-6879(90)85008-C] [PMID: 2199796]
[18]
Grodberg J, Dunn JJ. ompT encodes the Escherichia coli outer membrane protease that cleaves T7 RNA polymerase during purification. J Bacteriol 1988; 170(3): 1245-53.
[http://dx.doi.org/10.1128/jb.170.3.1245-1253.1988] [PMID: 3277950]
[19]
Jia B, Jeon CO. High-throughput recombinant protein expression in Escherichia coli: current status and future perspectives. Open Biol 2016; 6(8): 160196.
[http://dx.doi.org/10.1098/rsob.160196] [PMID: 27581654]
[20]
Loyevsky M, Mompoint F, Yikilmaz E, et al. Expression of a recombinant IRP-like Plasmodium falciparum protein that specifically binds putative plasmodial IREs. Mol Biochem Parasitol 2003; 126(2): 231-8.
[http://dx.doi.org/10.1016/S0166-6851(02)00278-5] [PMID: 12615322]
[21]
Terpe K. Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 2006; 72(2): 211-22.
[http://dx.doi.org/10.1007/s00253-006-0465-8] [PMID: 16791589]
[22]
Taylor RG, Walker DC, McInnes RR. E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing. Nucleic Acids Res 1993; 21(7): 1677-8.
[http://dx.doi.org/10.1093/nar/21.7.1677] [PMID: 8479929]
[23]
Anton BP, Raleigh EAJGA. Complete genome sequence of NEB 5-alpha, a derivative of Escherichia coli K-12 DH5α. Genome Announc 2016; 4(6): e01245-16.
[http://dx.doi.org/10.1128/genomeA.01245-16] [PMID: 27834703]
[24]
Song Y, Lee B-R, Cho S, et al. Determination of single nucleotide variants in Escherichia coli DH5α by using short-read sequencing. FEMS Microbiol Lett 2015; 362(11): fnv073.
[http://dx.doi.org/10.1093/femsle/fnv073] [PMID: 25934703]
[25]
Lobstein J, Emrich CA, Jeans C, Faulkner M, Riggs P, Berkmen M. SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm. Microb Cell Fact 2012; 11(1): 1-6.
[http://dx.doi.org/10.1186/1475-2859-11-56] [PMID: 22569138]
[26]
Ren G, Ke N, Berkmen M. Use of the shuffle strains in production of proteins. Current Protocols in Protein Science 2016; 85: 5.26.1-5.26.21.
[http://dx.doi.org/10.1002/cpps.11]
[27]
Gon S, Faulkner MJ, Beckwith J. In vivo requirement for glutaredoxins and thioredoxins in the reduction of the ribonucleotide reductases of Escherichia coli. Antioxid Redox Signal 2006; 8(5-6): 735-42.
[http://dx.doi.org/10.1089/ars.2006.8.735] [PMID: 16771665]
[28]
Bullock W. XL1-Blue: A high efficiency plasmid transforming recA Escherichia coli strain with beta- galactosidase selection. Biotechniques 1987; 5: 376-9.
[29]
Morowvat MH, Babaeipour V, Rajabi-Memari H, Vahidi H, Maghsoudi N. Overexpression of recombinant human beta interferon (rhINF-β) in periplasmic space of Escherichia coli. Iran J Pharm Res 2014; 13(Suppl.): 151-60.
[PMID: 24711841]
[30]
Girish V, Vijayalakshmi A. Affordable image analysis using NIH Image/ImageJ. Indian J Cancer 2004; 41(1): 47.
[PMID: 15105580]
[31]
Knipp M, Vasák M. A colorimetric 96-well microtiter plate assay for the determination of enzymatically formed citrulline. Anal Biochem 2000; 286(2): 257-64.
[http://dx.doi.org/10.1006/abio.2000.4805] [PMID: 11067748]
[32]
Noh EJ, Kang SW, Shin YJ, et al. Arginine deiminase enhances dexamethasone-induced cytotoxicity in human T-lymphoblastic leukemia CCRF-CEM cells. Int J Cancer 2004; 112(3): 502-8.
[http://dx.doi.org/10.1002/ijc.20435] [PMID: 15382078]
[33]
Liu S, Pritchard GG, Hardman MJ, Pilone GJ. Occurrence of arginine deiminase pathway enzymes in arginine catabolism by wine lactic Acid bacteria. Appl Environ Microbiol 1995; 61(1): 310-6.
[http://dx.doi.org/10.1128/aem.61.1.310-316.1995] [PMID: 16534912]
[34]
Ensor CM, Holtsberg FW, Bomalaski JS, Clark MA. Pegylated arginine deiminase (ADI-SS PEG20,000 mw) inhibits human melanomas and hepatocellular carcinomas in vitro and in vivo. Cancer Res 2002; 62(19): 5443-50.
[PMID: 12359751]
[35]
Han R-Z, Xu G-C, Dong J-J, Ni Y. Arginine deiminase: recent advances in discovery, crystal structure, and protein engineering for improved properties as an anti-tumor drug. Appl Microbiol Biotechnol 2016; 100(11): 4747-60.
[http://dx.doi.org/10.1007/s00253-016-7490-z] [PMID: 27087524]
[36]
Yang TS, Lu SN, Chao Y, et al. A randomised phase II study of pegylated arginine deiminase (ADI-PEG 20) in Asian advanced hepatocellular carcinoma patients. Br J Cancer 2010; 103(7): 954-60.
[http://dx.doi.org/10.1038/sj.bjc.6605856] [PMID: 20808309]
[37]
Ni Y, Schwaneberg U, Sun Z-H. Arginine deiminase, a potential anti-tumor drug. Cancer Lett 2008; 261(1): 1-11.
[http://dx.doi.org/10.1016/j.canlet.2007.11.038] [PMID: 18179862]
[38]
Shen L-J, Shen W-C. Drug evaluation: ADI-PEG-20- a PEGylated arginine deiminase for arginine-auxotrophic cancers. Curr Opin Mol Ther 2006; 8(3): 240-8.
[PMID: 16774044]
[39]
Zhu L, Verma R, Roccatano D, Ni Y, Sun ZH, Schwaneberg U. A potential antitumor drug (arginine deiminase) reengineered for efficient operation under physiological conditions. ChemBioChem 2010; 11(16): 2294-301.
[http://dx.doi.org/10.1002/cbic.201000458] [PMID: 20954230]
[40]
de Moura WAF, Schultz L, Breyer CA, et al. Functional and structural evaluation of the antileukaemic enzyme L-asparaginase II expressed at low temperature by different Escherichia coli strains. Biotechnol Lett 2020; 42(11): 2333-44.
[http://dx.doi.org/10.1007/s10529-020-02955-5] [PMID: 32638188]
[41]
Rai V, Upmanyu V, Mohd G, et al. Comparing the efficiency of different Escherichia coli strains in producing recombinant capsid protein of porcine circovirus type 2. Mol Cell Probes 2020; 52: 101556.
[http://dx.doi.org/10.1016/j.mcp.2020.101556] [PMID: 32126262]
[42]
Li S, Huang JC. Assessment of expression cassettes and culture media for different Escherichia coli strains to produce astaxanthin. Nat Prod Bioprospect 2018; 8(5): 397-403.
[http://dx.doi.org/10.1007/s13659-018-0172-z] [PMID: 29876754]
[43]
Makino T, Skretas G, Georgiou G. Strain engineering for improved expression of recombinant proteins in bacteria. Microb Cell Fact 2011; 10: 32.
[http://dx.doi.org/10.1186/1475-2859-10-32] [PMID: 21569582]
[44]
Fathi-Roudsari M, Akhavian-Tehrani A, Maghsoudi N. Comparison of three Escherichia coli strains in recombinant production of reteplase. Avicenna J Med Biotechnol 2016; 8(1): 16-22.
[PMID: 26855731]
[45]
Tegel H, Tourle S, Ottosson J, Persson A. Increased levels of recombinant human proteins with the Escherichia coli strain Rosetta(DE3). Protein Expr Purif 2010; 69(2): 159-67.
[http://dx.doi.org/10.1016/j.pep.2009.08.017] [PMID: 19733669]
[46]
Wang Z, Xiang Q, Wang G, Wang H, Zhang Y. Optimizing expression and purification of an ATP-binding gene gsiA from Escherichia coli k-12 by using GFP fusion. Genet Mol Biol 2011; 34(4): 661-8.
[http://dx.doi.org/10.1590/S1415-47572011005000043] [PMID: 22215971]
[47]
Doshi P, Bhargava P, Singh V, Pathak C, Joshi C, Joshi M. Escherichia coli strain engineering for enhanced production of serratiopeptidase for therapeutic applications. Int J Biol Macromol 2020; 160: 1050-60.
[http://dx.doi.org/10.1016/j.ijbiomac.2020.05.256] [PMID: 32497664]
[48]
Packiam KAR, Ramanan RN, Ooi CW, Krishnaswamy L, Tey BT. Stepwise optimization of recombinant protein production in Escherichia coli utilizing computational and experimental approaches. Appl Microbiol Biotechnol 2020; 104(8): 3253-66.
[http://dx.doi.org/10.1007/s00253-020-10454-w] [PMID: 32076772]
[49]
Xu Y-S, Du F, Li Z-J, et al. Regulating the T7 RNA polymerase expression in E. coli BL21 (DE3) to provide more host options for recombinant protein production. microb Cell Fact 2020; 20: 189.
[50]
Bomalaski J, Wu B-W. Methods of treatment with arginine deiminase. WO2013151568A1, 2013.
[51]
Ensor CM, Holtsberg FW, Clark MA. Mutated form of arginine deiminase. EP1278868B9, 2008.
[52]
Huang Y, Qin J, Fu X, Fan M, Wang Y, Wang Y. Arginine deiminase mutant and preparation and application therof. CN101812438A, 2014.

Rights & Permissions Print Export Cite as
© 2024 Bentham Science Publishers | Privacy Policy