Strategy for screening metagenomic resources for exocellulase activity using a robotic, high-throughput screening system

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Title
Strategy for screening metagenomic resources for exocellulase activity using a robotic, high-throughput screening system
Author(s)
Kyong-Cheol KoYunjon Han; Dae-Eun Cheong; Jong Hyun ChoiJae Jun Song
Bibliographic Citation
Journal of Microbiological Methods, vol. 94, no. 3, pp. 311-316
Publication Year
2013
Abstract
Exocellulases play a key role in cleaving the accessible ends of cellulose molecules to release soluble glucose and cellobiose. To date, there have been no screens for exocellulase owing to assay protocol limitations, the high cost of substrates, and low activity of exocellulases compared with endocellulases. This study is the first to demonstrate direct screening for exocellulase activity using a robotic, high-throughput screening (HTS) system. Cell growth in 96-well plates was measured by monitoring optical density over 11-14h at 37°C with agitation. Fluorescence of methylumbelliferyl groups released from 4-methylumbelliferyl-β-D-cellobioside was determined using a VICTOR3 microplate reader. This new HTS system enabled activity verification of more than 104 clones per day. As a result, we obtained four exocellulases clones (CelEx-SF301, CelEx-SF309, CelEx-BR12 and CelEx-BR15) from 29,006 metagenomic fosmid clones that had previously been prepared from sweet potato field soil microbes and rumen fluid. This powerful approach could be effectively applied to screen various metagenomic resources for new enzymes.
Keyword
Enzyme assayExocellulaseHigh-throughput screening (HTS)Metagenome
ISSN
0167-7012
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.mimet.2013.07.010
Type
Article
Appears in Collections:
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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