PMID- 26927878
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20170918
IS  - 1873-376X (Electronic)
IS  - 1570-0232 (Linking)
VI  - 1015-1016
DP  - 2016 Mar 15
TI  - Recovery and purification of a Kluyvermyces lactis beta-galactosidase by Mixed Mode 
      Chromatography.
PG  - 181-191
LID - S1570-0232(16)30070-8 [pii]
LID - 10.1016/j.jchromb.2016.01.053 [doi]
AB  - Mixed Mode Chromatography (MMC) is a potential separation technique that allows 
      simultaneous ionic and hydrophobic interactions between the adsorbent and the 
      adsorbate. The aim of this work was to assess the recovery and purification of a 
      Kluyveromyces lactis beta-galactosidase employing MMC. Protein precipitation and 
      dialysis were performed in order to concentrate the enzyme of interest and 
      eliminate cell debris and other interferences inherent in the fermentation 
      medium. The best conditions for both adsorption and desorption were attained by a 
      non-factorial Central Composite Experimental Design and employed in the 
      chromatographic runs with resin CAPTO MMC. Fermentation yielded mean values of 
      total enzyme concentration of 0.44 mg/mL, enzymatic activity (employing lactose 
      as a substrate) of 74 U/mL and specific activity of 168 U/mg. The Purification 
      Factor (PF) obtained was of 1.17. After precipitation and dialysis, the 
      subsequent chromatographic run resulted in recovery values ​​of 41.0 and 48.2% of 
      total protein concentration and enzymatic activity, respectively. SDS-PAGE 
      electrophoresis confirmed the purification evolution throughout the unit 
      operations employed, attesting the feasibility of the technique to obtain enzymes 
      with not only considerable degree of purity but also possessing high-added value.
CI  - Copyright (c) 2016 Elsevier B.V. All rights reserved.
FAU - Lima, Micael de Andrade
AU  - Lima MA
AD  - Universidade Federal do Ceara, Departamento de Engenharia Quimica, Campus do 
      Pici, S/N-Bloco 709-CEP, Ramal 206, 60455-760 Fortaleza, CE, Brazil.
FAU - Freitas, Maria de Fatima Matos de
AU  - Freitas MFM
AD  - Universidade Federal do Ceara, Departamento de Engenharia Quimica, Campus do 
      Pici, S/N-Bloco 709-CEP, Ramal 206, 60455-760 Fortaleza, CE, Brazil.
FAU - Goncalves, Luciana Rocha Barros
AU  - Goncalves LRB
AD  - Universidade Federal do Ceara, Departamento de Engenharia Quimica, Campus do 
      Pici, S/N-Bloco 709-CEP, Ramal 206, 60455-760 Fortaleza, CE, Brazil.
FAU - Silva Junior, Ivanildo Jose da
AU  - Silva Junior IJD
AD  - Universidade Federal do Ceara, Departamento de Engenharia Quimica, Campus do 
      Pici, S/N-Bloco 709-CEP, Ramal 206, 60455-760 Fortaleza, CE, Brazil. Electronic 
      address: ivanildo@gpsa.ufc.br.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160222
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (Bacterial Proteins)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - Adsorption
MH  - Bacterial Proteins/chemistry/*isolation & purification/metabolism
MH  - Chromatography, Liquid/*methods
MH  - Kluyveromyces/chemistry/*enzymology
MH  - Nonlinear Dynamics
MH  - beta-Galactosidase/chemistry/*isolation & purification/metabolism
OTO - NOTNLM
OT  - Adsorption
OT  - Biomolecule recovery and purification
OT  - Mixed Mode Chromatography
EDAT- 2016/03/02 06:00
MHDA- 2016/12/15 06:00
CRDT- 2016/03/02 06:00
PHST- 2015/09/01 00:00 [received]
PHST- 2016/01/28 00:00 [revised]
PHST- 2016/01/31 00:00 [accepted]
PHST- 2016/03/02 06:00 [entrez]
PHST- 2016/03/02 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - S1570-0232(16)30070-8 [pii]
AID - 10.1016/j.jchromb.2016.01.053 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Mar 15;1015-1016:181-191. 
      doi: 10.1016/j.jchromb.2016.01.053. Epub 2016 Feb 22.