| Biotechnology
Industry |
Microbial
Biotechnology |
| Electronic Journal of Biotechnology
ISSN: 0717-3458 |
Vol. 7 No. 2, Issue of August 15, 2004 |
| © 2004 by Pontificia Universidad Católica
de Valparaíso -- Chile |
Received January 7, 2004 / Accepted March 29, 2004 |
Catalytical properties of N-glycosylated Gluconacetobacter diazotrophicus
levansucrase produced in yeast
Luis Enrique Trujillo Toledo*
Division Plantas
Laboratorio Interacciones Planta-Microorganismos
Centro de Ingeniería Genética y Biotecnología
P.O. BOX 6162
C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: luis.trujillo@cigb.edu.cu
Raúl Gómez Riera
Division
Quimica Fisica
Centro de Ingeniería Genética y Biotecnología
C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: raul.gomez@cigb.edu.cu
Alexander
Banguela Castillo
Division
Plantas
Laboratorio Interacciones Planta-Microorganismos
Centro de Ingeniería Genética y Biotecnología
P.O. BOX 6162
C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: alexander.banguela@cigb.edu.cu
Melvis Soto Romero
Division
Plantas
Laboratorio Interacciones Planta-Microorganismos
Centro de Ingeniería Genética y Biotecnología
P.O. BOX 6162C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: melvis.soto@cigb.edu.cu
Juan Gabriel Arrieta
Sosa
Division
Plantas
Laboratorio Interacciones Planta-Microorganismos
Centro de Ingeniería Genética y Biotecnología
P.O. BOX 6162
C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: juan.arrieta@cigb.edu.cu
Lázaro Hernández García
Division
Plantas
Laboratorio Interacciones Planta-Microorganismos
Centro de Ingeniería Genética y Biotecnología
P.O. BOX 6162
C. Habana, Cuba
Tel: 53 7 2716967/6413 ext. 3117
Fax: 53 7 331779
E-mail: lazaro.hernandez@cigb.edu.cu
*Corresponding author
Financial support: This research was supported by a project grant
of the Cuban government # 3031254.
Keywords: catalytical
properties, fructo-oligosaccharides, fructosyltransferase, levansucrase,
N-glycosylation, Pichia pastoris.
Abbreviations:
FOS: fructo-oligosaccharides;
LsdA: Gluconacetobacter diazotrophicus levansucrase.
The influence of N-glycosylation
on the kinetic and catalytical properties of a bacterial fructosyltransferase
(LsdA) produced in Pichia pastoris was studied. The glycosylated
enzyme behaved similarly to non-glycosylated LsdA when substrate specificity,
fructo-oligosaccharide (FOS) production, sucrose hydrolysis or levan
formation reactions were carried out under different experimental
conditions. The kinetic parameters for native or yeast-expressed
LsdA determined at 60ºC, condition for the highest hydrolytic activity,
followed a conventional Michaelis-Menten kinetics. Synthase activity
of this levansucrase increased in water-restricted environments by
addition of salt or organic solvent to the reaction mixtures.
|
