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Characterization of Alginate Lyase Activity on Liquid, Gelled, and Complexed States of Alginate
Author(s) -
Breguet Véronique,
Stockar Urs von,
Marison Ian W.
Publication year - 2007
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp070150e
Subject(s) - enzyme , chemistry , ionic strength , glucuronic acid , hydrolysis , lyase , biochemistry , cell growth , calcium alginate , immobilized enzyme , chromatography , cell culture , kinetics , enzyme assay , biophysics , biology , organic chemistry , polysaccharide , aqueous solution , calcium , genetics , physics , quantum mechanics
A study of alginate lyase was carried out to determine if this enzyme could be used to remove alginate present in the core of alginate/poly‐ l ‐lysine (AG/PLL) microcapsules in order to maximize cell growth and colonization. A complete kinetic study was undertaken, which indicated an optimal activity of the enzyme at pH 7–8, 50 °C, in the presence of Ca 2+ . The buffer, not the ionic strength, influenced the alginate degradation rate. Alginate lyase was also shown to be active on gelled forms of alginate, as well as on the AG/PLL complex constituting the membrane of microcapsules. Batch cultures of CHO cells in the presence of alginate showed a decrease of the growth rate by a factor of 2, although the main metabolic flux rates were not modified. The addition of alginate lyase to cell culture medium increased the doubling time 5–7‐fold and decreased the protein production rate, although cell viability was not affected. The addition of enzyme to medium containing alginate did not improve growth conditions. This suggests that alginate lyase is probably not suitable for hydrolysis of microcapsules in the presence of cells, in order to achieve high cell density and high productivity. However, the high activity may be useful for releasing cells from alginate beads or AG/PLL microcapsules.