Abstract
The fungus Pseudozyma antarctica produces a lipase (CalB) with broad substrate specificity, stability, high regio- and enantio-selectivity. It is active in non-aqueous organic solvents and at elevated temperatures. Hence, CalB is a robust biocatalyst for chemical conversions on an industrial scale. Here we report the in silico mining of public metagenomes and fungal genomes to discover novel lipases with high homology to CalB. The candidates were selected taking into account homology and conserved motifs criteria, as well as, phylogeny and 3D model analyses. The most promising candidate (PlicB) presented interesting structural properties. PlicB was expressed in a heterologous host, purified and partially characterized. Further experiments will allow finding novel catalytic properties with biotechnological interest.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Biocatalysis
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Cloning, Molecular
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Data Mining
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Enzyme Stability
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Fungal Proteins / chemistry*
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Fungal Proteins / genetics
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Fungal Proteins / metabolism
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Genome, Fungal*
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Hot Temperature
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Kinetics
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Lipase / chemistry*
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Lipase / genetics
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Lipase / metabolism
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Molecular Sequence Data
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Phylogeny
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Pichia / genetics
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Pichia / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Alignment
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Solvents
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Stereoisomerism
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Substrate Specificity
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Ustilaginales / chemistry
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Ustilaginales / classification
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Ustilaginales / enzymology*
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Ustilaginales / genetics*
Substances
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Fungal Proteins
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Recombinant Proteins
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Solvents
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Lipase
Grants and funding
This work was supported by the Spanish projects BIO2012-36372, RTC-2014-1777-3 and S2013/MAE-2907. J. Barriuso thanks the financial support from the JAE-DOC CSIC program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.