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 10(4) 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.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Ko KC,Han Y,Cheong DE,Choi JH,Song JJdoi
10.1016/j.mimet.2013.07.010subject
Has Abstractpub_date
2013-09-01 00:00:00pages
311-6issue
3eissn
0167-7012issn
1872-8359pii
S0167-7012(13)00227-3journal_volume
94pub_type
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