Cell-free pipeline for discovery of thermotolerant xylanases and endo-1,4-β-glucanases

Gagoski, Dejan, Shi, Zhenyu, Nielsen, Lars Keld, Vickers, Claudia E., Mahler, Stephen, Speight, Robert, Johnston, Wayne A. and Alexandrov, Kirill (2017) Cell-free pipeline for discovery of thermotolerant xylanases and endo-1,4-β-glucanases. Journal of Biotechnology, 259 191-198. doi:10.1016/j.jbiotec.2017.07.014

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Author Gagoski, Dejan
Shi, Zhenyu
Nielsen, Lars Keld
Vickers, Claudia E.
Mahler, Stephen
Speight, Robert
Johnston, Wayne A.
Alexandrov, Kirill
Title Cell-free pipeline for discovery of thermotolerant xylanases and endo-1,4-β-glucanases
Formatted title
Cell-free pipeline for discovery of thermotolerant xylanases and endo-1,4-β-glucanases
Journal name Journal of Biotechnology   Check publisher's open access policy
ISSN 1873-4863
0168-1656
Publication date 2017-10-01
Year available 2017
Sub-type Article (original research)
DOI 10.1016/j.jbiotec.2017.07.014
Open Access Status File (Author Post-print)
Volume 259
Start page 191
End page 198
Total pages 8
Place of publication Amsterdam, NX, Netherlands
Publisher Elsevier
Language eng
Subject 1305 Biotechnology
2402 Applied Microbiology and Biotechnology
Abstract The rapid expansion in the number of sequenced genomes and metagenomes provides an exceptional resource for mining of the enzymes with biotechnologically relevant properties. However, the majority of protein production and analysis methods are not sufficiently cost-efficient and scalable to experimentally verify the results of computational genomic mining. Here, we present a pipeline based on Leishmania tarentolae cell-free system that was used to characterize 30 putative thermostable endo-1,4-beta-glucanases and xylanases identified in public genomic databases. In order to analyse the recombinant proteins without purification, novel high-throughput assays for glucanase and xylanase activities were developed. The assays rely on solubilisation of labelled particulate substrates performed in multiwell plates. Using this approach both acidophilic and thermophilic enzymes were identified. The developed approach enables rapid discovery of new biotechnologically useful enzymes.
Keyword Cell-free
Discovery
Enzyme
Glucanase
Metagenomes
Pipeline
Xylanase
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID APP1037320
DP1094080
LP120100162
Institutional Status UQ

 
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