TY - JOUR
T1 - A new mechanism and kinetic model for the enzymatic synthesis of short-chain fructooligosaccharides from sucrose
AU - Vega, Roberto
AU - Zuniga-Hansen, M. E.
N1 - Funding Information:
This research was supported financially by the CREAS . In addition, we acknowledge financial support (scholarship and purchase of software) from the Pontificia Universidad Católica de Valparaiso and CONICYT (scholarship 24100148 ) for R. Vega. We are very grateful to Matias Berndt (Dimerco Comercial Ltda., Chile) for the enzyme sample.
PY - 2014/1/15
Y1 - 2014/1/15
N2 - A kinetic model based on a ping-pong mechanism was developed under the steady-state hypothesis to account for the short-chain fructooligosaccharides (sc-FOS) synthesis using the commercial cellulolytic enzyme preparation, Rohapect CM. This new mechanism takes into account the interactions between the enzyme species and potential substrates (sucrose and sc-FOS) as a single complex reaction, allowing a better understanding of the reaction kinetics.The initial reaction rate laws appropriately describe the kinetic profiles of the examined substrates. Whereas sucrose exhibited Michaelis-Menten behavior with substrate inhibition, 1-kestose and nystose followed Michaelis-Menten and sigmoid enzyme kinetics. In addition, the enzyme was competitively inhibited by glucose and exhibited significant hydrolytic activity in the presence of nystose.The overall model was simultaneously fitted to experimental data from three initial sucrose concentrations (0.5, 1.5 and 2.1. M) using a multi-response regression with kinetic parameters that have biochemical relevance and are independent of the enzyme concentration. According to the model, sucrose acts almost exclusively as a fructosyl donor substrate. The mathematical development described herein is expected to be suitable for modeling similar enzymatic reaction systems.
AB - A kinetic model based on a ping-pong mechanism was developed under the steady-state hypothesis to account for the short-chain fructooligosaccharides (sc-FOS) synthesis using the commercial cellulolytic enzyme preparation, Rohapect CM. This new mechanism takes into account the interactions between the enzyme species and potential substrates (sucrose and sc-FOS) as a single complex reaction, allowing a better understanding of the reaction kinetics.The initial reaction rate laws appropriately describe the kinetic profiles of the examined substrates. Whereas sucrose exhibited Michaelis-Menten behavior with substrate inhibition, 1-kestose and nystose followed Michaelis-Menten and sigmoid enzyme kinetics. In addition, the enzyme was competitively inhibited by glucose and exhibited significant hydrolytic activity in the presence of nystose.The overall model was simultaneously fitted to experimental data from three initial sucrose concentrations (0.5, 1.5 and 2.1. M) using a multi-response regression with kinetic parameters that have biochemical relevance and are independent of the enzyme concentration. According to the model, sucrose acts almost exclusively as a fructosyl donor substrate. The mathematical development described herein is expected to be suitable for modeling similar enzymatic reaction systems.
KW - Enzyme biocatalysis
KW - Fructooligosaccharides
KW - Kinetic parameters
KW - Modeling
KW - Multi-response regression
KW - Sucrose
UR - http://www.scopus.com/inward/record.url?scp=84890043907&partnerID=8YFLogxK
U2 - 10.1016/j.bej.2013.11.012
DO - 10.1016/j.bej.2013.11.012
M3 - Artículo
AN - SCOPUS:84890043907
SN - 1369-703X
VL - 82
SP - 158
EP - 165
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
ER -