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Efficiency of Ionic Liquids–Based Aqueous Two-phase Electrophoresis for Partition of Cytochrome c

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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.

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Funding

This work was supported by the UCSI University Pioneer Scientist Incentive Funds under grant numbers [PROJ-In-FAS-027 and PROJ-2019-In-FAS-064] and Taiwan’s Ministry of Science and Technology under grant number [MOST 107-2621-M-155-001].

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Authors and Affiliations

  1. Faculty of Applied Sciences, UCSI University, UCSI Heights, Cheras, 56000, Kuala Lumpur, Malaysia

    Phei Er Kee, Hip Seng Yim & Hui-Suan Ng

  2. Biorefinery and Bioprocess Engineering Laboratory, Department of Chemical Engineering and Materials Science, Yuan Ze University, Chungli, Taoyuan, 320, Taiwan

    Phei Er Kee & John Chi-Wei Lan

  3. Bioprocess Technology, School of Industrial Technology, Universiti Sains Malaysia, 11800, Penang, Malaysia

    Joo Shun Tan

  4. School of Engineering, Taylor’s University, Lakeside Campus, No.1, Jalan Taylor’s, 47500, Subang Jaya, Selangor, Malaysia

    Yin Hui Chow

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  1. Phei Er Kee
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Correspondence to John Chi-Wei Lan or Hui-Suan Ng.

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Kee, P.E., Lan, J.CW., Yim, H.S. et al. Efficiency of Ionic Liquids–Based Aqueous Two-phase Electrophoresis for Partition of Cytochrome c. Appl Biochem Biotechnol 191, 376–386 (2020). https://doi.org/10.1007/s12010-019-03202-y

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  • DOI: https://doi.org/10.1007/s12010-019-03202-y

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