Process Biotechnology |
|
Animal Biotechnology |
| Electronic Journal of Biotechnology ISSN: 0717-3458 |
Vol.
12 No. 2, Issue of April 15, 2009 |
| © 2009 by Pontificia Universidad Católica
de Valparaíso -- Chile |
Received October 16, 2008 / Accepted January 12, 2009 |
| DOI: 10.2225/vol12-issue2-fulltext-10 |
|
Measuring β-Galactosidase activity
at pH 6 with a differential pH sensor
Cristian
Acevedo*#
Biotechnology Center
Universidad Técnica Federico Santa María
Av. España 1680
Valparaíso, Chile
Tel: 56 2 7184526
Fax: 56 2 7764080
E-mail: cristian.acevedo@usach.cl
Matthias H. Stach
Institute of Bioprocess Engineering
Friedrich-Alexander University of
Erlangen-Nuremberg
Paul Gordan Str. 3
91052 Erlangen, Germany
Anette Amtmann
Institute
of Bioprocess Engineering
Friedrich-Alexander University of Erlangen-Nuremberg
Paul Gordan Str. 3
91052 Erlangen, Germany
Manuel E. Young
Biotechnology Center
Universidad Técnica Federico Santa
María
Av. España 1680
Valparaíso, Chile
Juan G. Reyes
Chemistry Institute
Pontificia Universidad Católica de
Valparaíso
Av. Brasil 2950
Valparaíso, Chile
Holger Huebner
Institute of Bioprocess Engineering
Friedrich-Alexander University of
Erlangen-Nuremberg
Paul Gordan Str. 3
91052 Erlangen, Germany
Rainer Buchholz
Institute of Bioprocess Engineering
Friedrich-Alexander University of
Erlangen-Nuremberg
Paul Gordan Str. 3
91052 Erlangen, Germany
*Corresponding author
Financial support: FONDEF
Grant (DO2I1009), MECESUP Grant (UCV0206) and Doctoral Scholarship
for Cristian Acevedo (CONICYT D-21050588).
Present
address: #Departamento de Ciencia y Tecnología
de los Alimentos, Facultad Tecnológica, Universidad de
Santiago de Chile. Avenida Ecuador 3769, Estación Central,
Santiago, Chile.
Keywords: β-Galactosidase,
delta milli pH, differential pH.
Abbreviations: |
β-Gal: β-Galactosidase
β-Gal/pH6:
β-Galactosidase activity at pH 6
HK:
hexokinase
K:
Michaelis-Menten constant
t:
time
v:
enzymatic velocity
VMax:
maximum enzymatic velocity |
The β-Galactosidase activity at
pH 6 is used as a cellular marker to identify senescent cell cultures. The
classic method to identify this enzymatic activity is using cytochemical
staining with X-Gal after 16 hrs. In this work, a differential pH sensor was used
to measure β-Galactosidase activity at pH 6. The measurement is easy and
only takes 3 min.
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