Three-phase partitioning as a rapid and efficient method for purification of invertase from tomato


Özer B., Akardere E., Celem E. B. , Onal S.

BIOCHEMICAL ENGINEERING JOURNAL, cilt.50, ss.110-115, 2010 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 50
  • Basım Tarihi: 2010
  • Doi Numarası: 10.1016/j.bej.2010.04.002
  • Dergi Adı: BIOCHEMICAL ENGINEERING JOURNAL
  • Sayfa Sayıları: ss.110-115

Özet

Three-phase partitioning (TPP), a technique used in protein purification, was used to purify invertase from tomato (Lycopersicon esculentum). The method consists of simultaneous addition of ammonium sulfate and t-butanol to the crude enzyme extract in order to obtain the three phases. Different parameters (ammonium sulfate saturation, crude extract to t-butanol ratio and pH) essential for the extraction and purification of invertase were optimized to get highest purity fold and yield. It was seen that, 50% (w/v) ammonium sulfate saturation with 1:1 (v/v) ratio of crude extract to t-butanol at pH 4.5 gave 8.6-fold purification with 190% activity recovery of invertase in a single step. Finally, the purified enzyme was also characterized and the general biochemical properties were determined. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of enzyme showed considerable purification and its molecular weight was nearly found to be as 20 kDa. This work shows that, TPP is a simple, quick and economical technique for purification of invertases. (C) 2010 Elsevier B.V. All rights reserved.