Use of Triton X - 114 aqueous two phase system for recovery of mushroom (Agaricus bisporus) polyphenoloxidase

Download

index.pdf

Date

2003

Author

Beşel, Elif

Metadata

Show full item record

Item Usage Stats

128
views

0
downloads

Mushroom (Agaricus bisporus) polyphenoloxidase (PPO) (EC 1.14.18.1) was isolated and purified using an aqueous two phase system composed of octyl phenol (ethyleneglycol) 7-8 ether (Triton X-114/TX-114). TX-114 is a non-ionic surfactant which thermoseparates in water and forms an aqueous two phase system with a surfactant-depleted top phase and a surfactant-enriched bottom phase. Critical micelle concentration (CMC) and cloud point of the surfactant are 0.17 mM and 22ðC respectively. The partitioning behavior of mushroom PPO in water/TX-114 aqueous two-phase systems was studied and the effects of TX-114 concentration, ionic strength, pH, temperature and crude extract preparation on PPO partitioning were studied. PPO generally partitioned to the surfactant-depleted top phase; whereas many other hydrophobic proteins and molecules partitioned to the surfactantenriched bottom phase. When two-step ultrafiltration was used as a pretreatment, complete enzyme recovery was achieved with all studied TX-114 concentrations. Moreover, about 5 fold purification was achieved by using 8% TX-114. The purification increased to 10 fold by using polyvinylpolypyrolidone (PVPP) at pH 7.0 with a recovery of 72%. However changing pH from 7.0 to 6.0 increased the purification factor and enzyme recovery to 15 fold and 100%, respectively. Addition of potassium or sodium salts caused PPO molecules to partition in the surfactantenriched bottom phase. Finally, crude enzyme can be concentrated besides being purified and recovered by doing aqueous two-phase separation at room temperature.

Subject Keywords

Separation (Technology), Phase partition, Aqueous two phase separation, Polyphenoloxidase, Triton X-114, Thermoseparation, Purification

URI

http://etd.lib.metu.edu.tr/upload/1262971/index.pdf
https://hdl.handle.net/11511/13679

Collections

Graduate School of Natural and Applied Sciences, Thesis

Suggestions

OpenMETU
Core

Use of Specifically Tailored Chelating Polymers for Boron Removal from Aqueous Solutions by Polymer Enhanced Ultrafiltration Doganay, Ceren Oktar; Ozbelge, H. Onder; Bicak, Niyazi; AYDOĞAN, NİHAL; Yılmaz, Levent (Informa UK Limited, 2011-01-01) Two selective functional polychelatogens, namely hydroxyethylaminoglycerol functioned poly(glycidylmethacrylate (PNS) and Poly(4-Vinyl-1,3-dioxalan-2-one-co-vinyl acetate) (COP) were synthesized and utilized in polymer-enhanced ultrafiltration (PEUF) to remove boron from the aqueous solution. The effect of operating parameters on boron retention was investigated. PNS was synthesized in three molecular weights to observe the effect of molecular weight in borate retention. Increase in pH increased boron reten...
Development of alumina supported ternary mixed matrix membranes for separation of H-2/light-alkane mixtures Topuz, Berna; Yılmaz, Levent; Kalıpçılar, Halil (2012-10-01) Ternary component mixed matrix membrane was prepared from PES, SAPO-34 and 2-hydroxy 5-methyl aniline on a macroporous alumina disk by the solvent evaporation method in order to investigate the effect of existence of an inorganic support. The membrane and its pure PES/Alumina counterpart were characterized by single gas permeability measurements of H-2, CH4, C2H6 and C3H8. The corresponding H-2/CH4 selectivities of membranes were 71.3 and 41. The membranes were also used to separate equimolar mixtures of H-...
Effect of synthesis media pH and gel separation technique on properties of copper incorporated SBA-15 catalyst Akti, Filiz; Balci, Suna; Doğu, Timur (2019-10-01) Cu-SBA-15 catalysts were synthesized hydrothermally under acidic and slightly higher acidic conditions via solid separation by either gel filtration or evaporation. XRD (X-ray diffraction) patterns and HRTEM (high resolution transmission electron microscopy) images showed that metal loading caused little changes in the ordered structure of SBA-15. Characteristic structural SBA-15's bonds and acid sites were observed in FTIR (Fourier transform infrared) spectrums and synthesis under high pH was created chang...
Use of purified and modified bentonites in linear low-density polyethylene/organoclay/compatibilizer nanocomposites Seyidoglu, Tijen; Yılmazer, Ülkü (2012-05-05) Polyethylene-based ternary nanocomposites were prepared with different clay structures, obtained by the modification of purified Resadiye bentonite as the reinforcement, a random terpolymer of ethylene, butyl acrylate, and maleic anhydride with the trade name Lotader3210 as the compatibilizer, and linear low-density polyethylene (LLDPE) as the polymer matrix in an intensive batch mixer. The quaternary ammonium/phosphonium salts used for the modification of bentonite were dimethyldioctadecyl ammonium (DMDA) ...
Removal of methylene blue by new tunable aryl/alkyl ionic liquids/salts (TAAILs) from aqueous solution Özdemir, Melek (Informa UK Limited, 2019-10-16) The removal of methylene blue (MB) from aqueous solution has been investigated using new tunable 1-aryl-2-alkyl-3,5-dimethylpyrazolium hexafluorophosphate salts ([Ph(R)C(R?)pz][PF6], R: -Cl, -Br, -Me, -OMe, R?: -C2H5, or -C4H9) as an extractant and methylene chloride as a diluent. The influence of p-substituent and alkyl chain length on the removal efficiency of MB from aqueous solution was investigated. The obtained results show that MB was extracted from aqueous solution with high extraction efficiency up...

Citation Formats

E. Beşel, “Use of Triton X - 114 aqueous two phase system for recovery of mushroom (Agaricus bisporus) polyphenoloxidase,” M.S. - Master of Science, Middle East Technical University, 2003.