title: Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans.
abstract: The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values.
authors.affiliation: Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Gachibowli, Hyderabad, India.
authors.forename: Jogi
authors.initials: J
authors.lastname: Madhuprakash
authors.affiliation: Institute for Biology and Biotechnology of Plants, WWU, Münster University, Münster, Germany.
authors.forename: Nour Eddine
authors.initials: NE
authors.lastname: El Gueddari
authors.affiliation: Institute for Biology and Biotechnology of Plants, WWU, Münster University, Münster, Germany.
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Moerschbacher
authors.affiliation: Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Gachibowli, Hyderabad, India. Electronic address: arpsl@uohyd.ernet.in.
authors.forename: Appa Rao
authors.initials: AR
authors.lastname: Podile
mesh_terms: D000107:Acetylation; D018929:Cell Culture Techniques / Q000379:methods; D002688:Chitinases / Q000235:genetics / Q000378:metabolism*; D048271:Chitosan / Q000378:metabolism*; D006031:Glycosylation; D006868:Hydrolysis; D009154:Mutation; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D012275:Oryza / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016897:Respiratory Burst / Q000187:drug effects; D012705:Serratia / Q000201:enzymology*
chemical_list: D009844:Oligosaccharides; D048271:Chitosan; D002688:Chitinases
keywords: Chitinases; Chitosan oligosaccharides; Hydrolysis; SpChiD; Transglycosylation