A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria

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A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria
Kyong-Cheol Ko; J H Lee; Yunjon HanJong Hyun ChoiJae Jun Song
Bibliographic Citation
Biochemical and Biophysical Research Communications, vol. 441, no. 3, pp. 567-572
Publication Year
Metagenomic resources representing ruminal bacteria were screened for novel exocellulases using a robotic, high-throughput screening system, the novel CelEx-BR12 gene was identified and the predicted CelEx-BR12 protein was characterized. The CelEx-BR12 gene had an open reading frame (ORF) of 1140 base pairs that encoded a 380-amino-acid-protein with a predicted molecular mass of 41.8kDa. The amino acid sequence was 83% identical to that of a family 5 glycosyl hydrolase from Prevotella ruminicola 23. Codon-optimized CelEx-BR12 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The Michaelis-Menten constant (Km value) and maximal reaction velocity (Vmax values) for exocellulase activity were 12.92μM and 1.55×10-4μmolmin-1, respectively, and the enzyme was optimally active at pH 5.0 and 37°C. Multifunctional activities were observed against fluorogenic and natural glycosides, such as 4-methylumbelliferyl-β-d-cellobioside (0.3Umg-1), CMC (105.9Umg-1), birch wood xylan (132.3Umg-1), oat spelt xylan (67.9Umg-1), and 2-hydroxyethyl-cellulose (26.3Umg-1). Based on these findings, we believe that CelEx-BR12 is an efficient multifunctional enzyme as endocellulase/exocellulase/xylanase activities that may prove useful for biotechnological applications.
Cellulolytic enzymeEnzyme assayMetagenomeMultifunctional enzyme
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Ochang Branch Institute > Division of National Bio-Infrastructure > Korea Preclinical Evaluation Center > 1. Journal Articles
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
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