Enzymatic production of defined chitosan oligomers with a specific pattern of acetylation using a combination of chitin oligosaccharide deacetylases.
Chitin and chitosan oligomers have diverse biological activities with potentially valuable applications in fields like medicine, cosmetics, or agriculture. These properties may depend not only on the degrees of polymerization and acetylation, but also on a specific pattern of acetylation (PA) that cannot be controlled when the oligomers are produced by chemical hydrolysis. To determine the influence of the PA on the biological activities, defined chitosan oligomers in sufficient amounts are needed. Chitosan oligomers with specific PA can be produced by enzymatic deacetylation of chitin oligomers, but the diversity is limited by the low number of chitin deacetylases available. We have produced specific chitosan oligomers which are deacetylated at the first two units starting from the non-reducing end by the combined use of two different chitin deacetylases, namely NodB from Rhizobium sp. GRH2 that deacetylates the first unit and COD from Vibrio cholerae that deacetylates the second unit starting from the non-reducing end. Both chitin deacetylases accept the product of each other resulting in production of chitosan oligomers with a novel and defined PA. When extended to further chitin deacetylases, this approach has the potential to yield a large range of novel chitosan oligomers with a fully defined architecture.
Institute of Plant Biology and Biotechnology, Westphalian Wilhelm's-University Münster, Schlossplatz 8, 48143 Münster, Germany.
Stefanie Nicole
SN
Hamer
Institute of Plant Biology and Biotechnology, Westphalian Wilhelm's-University Münster, Schlossplatz 8, 48143 Münster, Germany.
Stefan
S
Cord-Landwehr
Laboratory of Biochemistry, Institut Químic de Sarrià (IQS), Universitat Ramon Llull (URL), Via Augusta 390, E-08017 Barcelona, Spain.
Xevi
X
Biarnés
Laboratory of Biochemistry, Institut Químic de Sarrià (IQS), Universitat Ramon Llull (URL), Via Augusta 390, E-08017 Barcelona, Spain.
Antoni
A
Planas
Bio Base Europe Pilot Plant, Rodenhuizekaai 1, 9042 Ghent, Belgium.
Hendrik
H
Waegeman
Institute of Plant Biology and Biotechnology, Westphalian Wilhelm's-University Münster, Schlossplatz 8, 48143 Münster, Germany.
Bruno Maria
BM
Moerschbacher
Institute of Plant Biology and Biotechnology, Westphalian Wilhelm's-University Münster, Schlossplatz 8, 48143 Münster, Germany.
Stephan
S
Kolkenbrock
D000107:Acetylation; D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002384:Catalysis; D020134:Catalytic Domain; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012231:Rhizobium / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity
D011994:Recombinant Proteins; D048271:Chitosan; D000581:Amidohydrolases; C019866:chitin deacetylase