Affiliations 

  • 1 Faculty of Applied Sciences, UCSI University, UCSI Heights, Cheras, 56000, Kuala Lumpur, Malaysia
  • 2 Biorefinery and Bioprocess Engineering Laboratory, Department of Chemical Engineering and Materials Science, Yuan Ze University, Chungli, Taoyuan, 320, Taiwan. lanchiwei@saturn.yzu.edu.tw
  • 3 Bioprocess Technology, School of Industrial Technology, Universiti Sains Malaysia, 11800, Penang, Malaysia
  • 4 School of Engineering, Taylor's University, Lakeside Campus, No.1, Jalan Taylor's, 47500, Subang Jaya, Selangor, Malaysia
  • 5 Faculty of Applied Sciences, UCSI University, UCSI Heights, Cheras, 56000, Kuala Lumpur, Malaysia. GrraceNg@ucsiuniversity.edu.my
Appl Biochem Biotechnol, 2020 May;191(1):376-386.
PMID: 31907777 DOI: 10.1007/s12010-019-03202-y

Abstract

Cytochrome c is a small water-soluble protein that is abundantly found in the mitochondrial intermembrane space of microorganism, plants and mammalians. Ionic liquids (ILs)-based aqueous two-phase electrophoresis system (ATPES) was introduced in this study to investigate the partition efficiency of cytochrome c to facilitate subsequent development of two-phase electrophoresis for the separation of cytochrome c from microbial fermentation. The 1-Hexyl-3-methylimidazolium bromide, (C6mim)Br and potassium citrate salt were selected as the phase-forming components. Effects of phase composition; position of electrodes; pH and addition of neutral salt on the partition efficiency of cytochrome c in the ATPES were evaluated. Highest partition coefficient (K = 179.12 ± 0.82) and yield of cytochrome c in top phase (YT = 99.63% ± 0.00) were recorded with IL/salt ATPES composed of 30% (w/w) (C6mim)Br and 20% (w/w) potassium citrate salt of pH 7 and 3.0% (w/w) NaCl addition with anode at the bottom phase and cathode at the top phase. The SDS-PAGE profile revealed that cytochrome c with a molecular weight of 12 kDa was preferably partitioned to the IL-rich top phase. Present findings suggested that the single-step ATPES is a potential separation approach for the recovery of cytochrome c from microbial fermentation. Graphical Abstract.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.