Optimization of S-layer protein purification from Deinococcus radiodurans strain R1

Authors

  • Abdolirad, Maryam Department of Bioscience and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran
  • Alijanianzadeh, Mahdi Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Karaj, Iran
  • Jalalvand, Alireza Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran
  • Khalilzadeh, Rasoul Department of Bioscience and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran
Abstract:

S-layer proteins of Deinococcus radiodurans are the best self-assemble systems among other proteins that have an essential role in the fabrication of nanowires. Therefore, the purification of these proteins is necessary. The purpose of this research was to optimize the purification of s-layer protein from D. radiodurans with the response surface method. The three factors of SDS concentration, incubation time and mass percent in five levels were considered, and 20 runs were designed by Design-Expert software with a central composite method. Each run includes microbe culture, mass cell preparation, microbe incubation in specific SDS concentration and time and mass percent, separation of the bacteria from detergent with a centrifuge at 5000g, sedimentation of s-layer proteins from detergent solution with a centrifuge at 20000g, determination of protein concentration, and protein purity by Bradford and SDS-PAGE methods, respectively. Finally, the data obtained were analyzed.  Analysis of the results demonstrated that at the 95% confidence level, the effect of the detergent concentration factor on the purified protein percent was more than other factors. The optimization results of factors are 5.64% SDS concentration, 7.33% mass percent, and 3 hours incubation time. At optimized conditions the protein concentration and purity percent were obtained 0.584 mg/ml and 47.61% respectively.  

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Isolation and Study of S-layer Nanostructure of Deinococcus Radiodurans R1

Crystalline surface layer proteins (S-layer proteins) have considerable potential for the crystalline arrays in biotechnology, biomimetics and nonlife applications, including areas such as microelectronics and molecular nanotechnology. The extensive application potential of surface layers in nanobiotechnology is according to the particular inherent attributes of the single molecular arrays cons...

full text

Protein splicing of the Deinococcus radiodurans strain R1 Snf2 intein.

Adjacent intein fragments fused to a Snf2/Rad54 helicase-related protein and Snf2/Rad54 helicase were reported for Deinococcus radiodurans R1, leading to the speculation that a frameshift was required for splicing or that trans splicing occurred. However, a type strain (ATCC 13939, RF18410) yielded a single protein that splices by the Ala1 protein splicing pathway, with splicing dependent on ad...

full text

Purification and characterization of DR_2577 (SlpA) a major S-layer protein from Deinococcus radiodurans

The protein DR_2577 is a major Surface layer component of the radio-resistant bacterium Deinococcus radiodurans. In the present study DR_2577 has been purified and its oligomeric profile characterized by means of size exclusion chromatography and gel electrophoresis. DR_2577 was found to be organized into three hierarchical orders characterized by monomers, stable dimers formed by the occurrenc...

full text

Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1.

The complete genome sequence of the radiation-resistant bacterium Deinococcus radiodurans R1 is composed of two chromosomes (2,648,638 and 412,348 base pairs), a megaplasmid (177,466 base pairs), and a small plasmid (45,704 base pairs), yielding a total genome of 3,284, 156 base pairs. Multiple components distributed on the chromosomes and megaplasmid that contribute to the ability of D. radiod...

full text

The IrrE protein of Deinococcus radiodurans R1 is a novel regulator of recA expression.

IRS24 is a DNA damage-sensitive strain of Deinococcus radiodurans strain 302 carrying a mutation in an uncharacterized locus designated irrE. Five overlapping cosmids capable of restoring ionizing radiation resistance to IRS24 were isolated from a genomic library. The ends of each cloned insert were sequenced, and these sequences were used to localize irrE to a 970-bp region on chromosome I of ...

full text

Correction: Preserving Genome Integrity: The DdrA Protein of Deinococcus radiodurans R1

The bacterium Deinococcus radiodurans can withstand extraordinary levels of ionizing radiation, reflecting an equally extraordinary capacity for DNA repair. The hypothetical gene product DR0423 has been implicated in the recovery of this organism from DNA damage, indicating that this protein is a novel component of the D. radiodurans DNA repair system. DR0423 is a homologue of the eukaryotic Ra...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 9  issue 4

pages  267- 278

publication date 2023-03

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

No Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023