Production of YY Peptides in the Escherichia Coli Expression System Using Self-Induction Promoter

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Microbiology, School of Basic Sciences, Karaj Branch, Islamic Azad University, Karaj, Iran

2 Assistant Professor, Department of System Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

3 Assistant Professor, Department of Microbiology, School of Basic Sciences, Karaj Branch, Islamic Azad University, Karaj, Iran

Abstract

Background: Application of toxic inducers, i.e. isopropyl β-D-1-thiogalactopyranoside (IPTG), for production of biopharmaceuticals has been prohibited by regulatory agencies. Here, we sought to evaluate the feasibility of a self-induced expression system for production of YY(3-36) peptide as an active pharmaceutical ingredient (API) in the Escherichia coli expression system.Methods: The sequence of self-induced promoter was obtained from Data banks, and synthetized in pUC18 vector. The nucleotide sequence of YY(3-36) was inserted downstream of the promoter, after the 22 residue of asparaginase II signal sequence. The expression of the peptide was compared with pET21 harboring the same construct under isopropyl β-D-1-thiogalactopyranoside induction in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Findings: The expression of target peptide was comparable with isopropyl β-D-1-thiogalactopyranoside (1 mM) expressed peptide.Conclusion: Our results demonstrate the ease and economical application of used self-induced promoter as an alternative system in production of high value added biopharmaceuticals.

Keywords

References

  1. Bashyam MD, Tyagi AK. Identification and analysis of "extended -10" promoters from mycobacteria. J Bacteriol 1998; 180(9): 2568-73.
  2. Ojala V, Laitalainen J, Jalasvuori M. Fight evolution with evolution: plasmid-dependent phages with a wide host range prevent the spread of antibiotic resistance. Evol Appl 2013; 6(6): 925-32.
  3. Campbell JL, Richardson CC, Studier FW. Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci USA 1978; 75(5): 2276-80.
  4. Adham SA, Rodriguez S, Ramos A, Santamaria RI, Gil JA. Improved vectors for transcriptional/translational signal screening in corynebacteria using the melC operon from Streptomyces glaucescens as reporter. Arch Microbiol 2003; 180(1): 53-9.
  5. Davanloo P, Rosenberg AH, Dunn JJ, Studier FW. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci USA 1984; 81(7): 2035-9.
  6. Backman K, Ptashne M, Gilbert W. Construction of plasmids carrying the cI gene of bacteriophage lambda. Proc Natl Acad Sci USA 1976; 73(11): 4174-8.
  7. Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, et al. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 1977; 2(2): 95-113.
  8. Carter AD, Morris CE, McAllister WT. Revised transcription map of the late region of bacteriophage T7 DNA. J Virol 1981; 37(2): 636-42.
  9. Fazen CH, Kahkoska AR, Doyle RP. Expression and purification of human PYY(3-36) in Escherichia coli using a His-tagged small ubiquitin-like modifier fusion. Protein Expr Purif 2012; 85(1): 51-9.
  10. Choi JH, Lee SY. Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl Microbiol Biotechnol 2004; 64(5): 625-35.
  11. Ehira S, Teramoto H, Inui M, Yukawa H. Regulation of Corynebacterium glutamicum heat shock response by the extracytoplasmic-function sigma factor SigH and transcriptional regulators HspR and HrcA. J Bacteriol 2009; 191(9): 2964-72.
  12. Mahboobi M, Sedighian H, Hedayati CH M, Bambai B, Esmaeil Soofian S, et al. Applying bioinformatic tools for modeling and modifying type II E. coli l-asparginase to present a better therapeutic agent/drug for acute lymphoblastic leukemia. Int J Cancer Manag. 2017; 10(3): e5785.
  13. Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 1980; 65(1): 499-560.
  14. McAllister WT, Morris C, Rosenberg AH, Studier FW. Utilization of bacteriophage T7 late promoters in recombinant plasmids during infection. J Mol Biol 1981; 153(3): 527-44.
  15. Heiss S, Hormann A, Tauer C, Sonnleitner M, Egger E, Grabherr R, et al. Evaluation of novel inducible promoter/repressor systems for recombinant protein expression in Lactobacillus plantarum. Microb Cell Fact 2016; 15: 50.
  16. Conway T, Creecy JP, Maddox SM, Grissom JE, Conkle TL, Shadid TM, et al. Unprecedented high-resolution view of bacterial operon architecture revealed by RNA sequencing. MBio 2014; 5(4): e01442-14.
Journal of Isfahan Medical School
Volume 36, Issue 468 - Serial Number 468
January and February 2018
Pages 118-123

Files

History

  • Receive Date: 26 September 2017
  • Revise Date: 19 April 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics