[en] Introduction. Discovering novel enzymes is of interest in both applied and basic science. Microbial enzymes, which are incredibly diverse and easy to produce, are increasingly sought by diverse approaches.
Literature. This review first distinguishes culture-based from culture-independent methods, detailing within each group the advantages and drawbacks of sequence- and function-based methods. It then discusses the main factors affecting the success of endeavors to identify novel enzymes through construction and functional screening of genomic or metagenomic libraries: the sampled environment, how DNA is extracted and processed, the vector used (plasmid, cosmid, fosmid, BAC, or shuttle vector), the host cell chosen from the available prokaryotic and eukaryotic ones and the main screening steps.
Conclusions. Library construction and screening can be tricky and requires expertise. Combining different strategies, such as working with cultivable and non-cultivable organisms, using sequence- and function-based approaches, or performing multihost screenings, is probably the best way to identify novel and diverse enzymes from an environmental sample.
Disciplines :
Microbiology
Author, co-author :
Martin, Marjolaine ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbiologie et génomique
Vandenbol, Micheline ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Microbiologie et génomique
Language :
English
Title :
The hunt for original microbial enzymes: an initiatory review on the construction and functional screening of (meta)genomic libraries
Alternative titles :
[fr] La chasse aux enzymes microbiennes originales: une synthèse bibliographique initiatique sur la construction et le criblage fonctionnel de banques (méta)génomiques
Publication date :
2016
Journal title :
Biotechnologie, Agronomie, Société et Environnement
ISSN :
1370-6233
eISSN :
1780-4507
Publisher :
Presses Agronomiques de Gembloux, Gembloux, Belgium
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