Potential application of aqueous two‐phase systems and three‐phase partitioning for the recovery of superoxide dismutase from a clarified homogenate of Kluyveromyces marxianus

Abstract

Superoxide dismutase (SOD; EC 1.15.1.1) is an antioxidant enzyme that represents the primary cellular defense against superoxide radicals and has interesting applications in the medical and cosmetic industries. In the present work, the partition behavior of SOD in aqueous two-phase systems (ATPS) (using a standard solution and a complex extract from Kluyveromyces marxianus as sample) was characterized on different types of ATPS (polymer-polymer, polymer-salt, alcohol-salt, and ionic liquid (IL)-salt). The systems composed of PEG 3350-potassium phosphate, 45% TLL, 0.5 M NaCl (315 U/mg, 87% recovery, and 15.1-fold purification) and t-butanol-20% ammonium sulfate (205.8 U/mg, 80% recovery and 9.8-fold purification), coupled with a subsequent 100 kDa ultrafiltration stage, allowed the design of a prototype process for the recovery and partial purification of the product of interest. The findings reported herein demonstrate the potential of PEG-salt ATPS for the potential recovery of SOD.

Ionic-Liquid-Mediated Extraction and Separation Processes for Bioactive Compounds: Past, Present, and Future Trends

“…Although IL-based ABS were among the two systems that allowed better enzyme recovery, the authors concluded that the PEG3350-potassium phosphate ABS was better in terms of SOD recovery, enzyme specific activity maintenance, and purification. 259 …”

“…Although IL-based ABS were among the two systems that allowed better enzyme recovery, the authors concluded that the PEG3350-potassium phosphate ABS was better in terms of SOD recovery, enzyme specific activity maintenance, and purification. 259 Further applications of IL-based ABS in the purification of enzymes include the extraction of wheat esterase from wheat extracts. 260 This enzyme can substitute the traditional acetylcholinesterase (AChE) extracted from animal blood or tissues.…”

“…To overcome these drawbacks, IL-based ABS are considered as an alternative path for a faster and more effective purification method. A variety of ABS (polymer–polymer, polymer-salt, alcohol-salt, and IL-salt) were studied for the SOD recovery by Simental-Martinez et al These authors studied the SOD partition behavior in ABS using a standard solution and also a complex extract obtained from the yeast Kluyveromyces marxianus. Their results indicated that the enzyme always partititioned into the IL-rich phase .…”

“…A variety of ABS (polymer–polymer, polymer-salt, alcohol-salt, and IL-salt) were studied for the SOD recovery by Simental-Martinez et al These authors studied the SOD partition behavior in ABS using a standard solution and also a complex extract obtained from the yeast Kluyveromyces marxianus. Their results indicated that the enzyme always partititioned into the IL-rich phase . Satisfactory purification results were achieved with an activity recovery yield of 91%, with 60% of the total protein being found in the same phase.…”

Partitioning in aqueous two‐phase systems: Analysis of strengths, weaknesses, opportunities and threats

“…One example is the successful pioneering work of Hayenga et al on chymosin purification [94]. Other examples include: (i) separation of plasmid DNA from mRNA and tRNA [73]; (ii) simultaneous extraction and purification of lectins from plant extracts [95]; (iii) purification of human growth hormone from a fermentation broth [18]; (iv) purification of recombinant human serum albumin from viscous fermentation feed [50]; (v) purification of enzymes such as superoxide dismutase [96] and laccases [58, 97] from fermentation feeds; and (vi) extraction and purification of retamycin [14] and tetracycline [98] direct from fermentation broth.…”