Browsing by Author "Marais, Charl"
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- ItemScreening, isolating and characterizing acetyl xylan esterase enzymes from a novel ecological niche(Stellenbosch : Stellenbosch University, 2015-03) Marais, Charl; Kossmann, Jens; George, Gavin M.; Stellenbosch University. Faculty of Agrisciences. Dept. of Genetics. Institute for Plant Biotechnology (IPB).ENGLISH ABSTRACT: Hemicellulose, a plant cell wall polysaccharide composed amongst other 6-carbon sugars of xylose, a 5-carbon fermentable sugar, has various side-chains which hinder breakdown by the hydrolytic enzyme xylanase. We executed a screen of a metagenomic library established from heat treated saw dust to isolate novel genes for plant cell wall esterases and isolated two clones that potentially could be utilised as acetyl xylan esterases: HEL12 and HEL37 from Citrobacter farmeri and Bacillus vallismortis respectively are confirmed short-chain acetyl xylan esterases (C2-C4) with an optimal temperature of 30°C and 35°C and pH 8.0. HEL12 (33kDa) and HEL37 (25kDa) are small, dextrous acetyl xylan esterases with HEL37: Km of 1.621mM for p-nitrophenyl acetate and Km of 3.571mM for the substrate p-nitrophenyl palmitate with a Vmax of 2.462 mMol/min/mg protein and Vmax of 0.4363 mMol/min/mg protein respectively. In addition HEL12 was active on other substrates with a Km of 1.321mM protein for p-nitrophenyl acetate and Km of 1.692mM for the substrate p-nitrophenyl butyrate with a Vmax of 3.812 and Vmax of 1.523 mMol/min/mg protein respectively. Both enzymes were assayed on various acetylated polymers including acetylated xylan to indicate their ability to hydrolyse plant lignocellulosic polymers.