Direct recovery of cyclodextringlycosyltransferase from Bacillus cereus using aqueous two-phase flotation

J Biosci Bioeng. 2015 Dec;120(6):684-9. doi: 10.1016/j.jbiosc.2015.04.013. Epub 2015 Jun 22.

Abstract

Purification of cyclodextrin glycosyl transferase (CGTase) from Bacillus cereus using polyethylene glycol (PEG)-potassium phosphates aqueous two-phase flotation (ATPF) system was studied in this paper. The effects of varying PEG molecular weight, tie-line length (TLL) value, volume ratio (VR), pH value, crude concentration and gas nitrogen flotation time were investigated. The optimal condition for purification of CGTase was attained at 18.0% (w/w) PEG 8000, 7.0% (w/w) potassium phosphates, VR of 3.0, 20% (w/w) crude load at pH 7, and 80 min nitrogen flotation time at a flow rate of 5 L/min. With this optimal condition, purification factor (PFT) of 21.8 and a yield (YT) of 97.1% were attained. CGTase was successfully purified in a single downstream processing step using the ATPF.

Keywords: Aqueous two-phase flotation; Bacillus cereus; Cyclodextringlycosyltransferase; Polyethylene glycol-salt system; Purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus cereus / enzymology*
  • Glucosyltransferases / chemistry*
  • Glucosyltransferases / isolation & purification*
  • Glucosyltransferases / metabolism
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Nitrogen
  • Phosphates / chemistry
  • Polyethylene Glycols / chemistry
  • Potassium Compounds / chemistry
  • Time Factors
  • Water / chemistry*

Substances

  • Phosphates
  • Potassium Compounds
  • Water
  • Polyethylene Glycols
  • potassium phosphate
  • Glucosyltransferases
  • cyclomaltodextrin glucanotransferase
  • Nitrogen