Details

Autor(en) / Beteiligte

• Vaikuntapu, Papa Rao
• Mallakuntla, Mohan Krishna
• Das, Subha Narayan
• Bhuvanachandra, Bhoopal
• Ramakrishna, Bellamkonda
• Nadendla, Sandhya Rani
• Podile, Appa Rao

Titel

Applicability of endochitinase of Flavobacterium johnsoniae with transglycosylation activity in generating long-chain chitooligosaccharides

Ist Teil von

• International journal of biological macromolecules, 2018-10, Vol.117, p.62-71

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2018

Quelle

Elsevier ScienceDirect Journals

Beschreibungen/Notizen

• Chitin and its derivatives are used for a variety of applications. Flavobacterium johnsoniae UW101 is an aerobic Gram-negative bacterium. Genome analysis of F. johnsoniae UW101 revealed the presence of 10 glycoside hydrolases (GHs) that may degrade or modify chitin. The gene encoding chitinase B (FjchiB), which encodes a single catalytic GH18 domain has been cloned and heterologously expressed in Escherichia coli. FjChiB was optimally active in 50 mM sodium citrate buffer (pH 6.0) at 40 °C. FjChiB was salt-tolerant and catalytically versatile, with substrate specificity towards 75% DDA (degree of de-acetylation) chitosan, followed by colloidal chitin. Chitotetraose (DP4) was the shortest of the oligomeric substrates used by FjChiB. The Km and Vmax values of FjChiB for colloidal chitin were 49.38 mg/ml and 11.2 nanokat mg−1, respectively. The overall catalytic efficiency (kcat/Km) of FjChiB was 1.40 × 103 mg−1 ml s−1. FjChiB exhibited transglycosylation (TG) with chitopentaose (DP5) and chitohexaose (DP6) substrates. The TG by FjChiB was fine-tuned by introducing a tryptophan (G106W) and asparagine (D148N) in the highly conserved catalytic groove and catalytic center, respectively. Hydrolytic products profile and homology modelling indicated that FjChiB is an endochitinase that holds promise for the conversion of chitin into useful products through both TG and/or hydrolysis. •Characterized FjChiB from Flavobacterium johnsoniae which is a unique and single modular salt-tolerant endochitinase•Chitobiose (DP2) and chitotriose (DP3) are not hydrolyzed by FjChiB.•FjChiB exhibited transglycosylation on chitopentaose (DP5) and chitohexaose (DP6).