To investigate the properties of a novel metagenome-derived member of the hormone-sensitive lipase family of lipolytic enzymes.
A forest soil metagenome-derived gene encoding an esterase (Est06) belonging to the hormone-sensitive lipase family of lipolytic enzymes was subcloned, heterologously expressed and characterized. Est06 is a polypeptide of 295 amino acids with a molecular mass of 31 kDa. The deduced protein sequence shares 61% similarity with a hypothetical protein from the marine symbiont Candidatus Entotheonella sp. TSY1. Purified Est06 exhibited high affinity for acyl esters with short-chain fatty acids, and showed optimum activity with p-nitrophenyl valerate (C5). Maximum enzymatic activity was at 50 °C and pH 7. Est06 exhibited high stability at moderate temperatures by retaining all of its catalytic activity below 30 °C over 13 days. Additionally, Est06 displayed high stability between pH 5 and 9. Esterase activity was not inhibited by metal ions or detergents, although organic solvents decreased activity.
The combination of Est06 properties place it among novel biocatalysts that have potential for industrial use including low temperature applications.
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We thank the DFG (German Science Foundation) for funding this work in the context of the graduate school ‘Die Bedeutung der Biodiversität für Stoffkreisläufe und biotische Interaktionen in temperaten Laubwäldern (GRK 1086)’. We thank Dr. Heiko Nacke for providing the lipolytic clone pLE06 used in this work, as well as Florian Jung and Jörn Lindemann for providing technical assistance.
Supplementary Table 1—Amino acid sequence similarities between Est06 and related lipolytic proteins.
Supplementary Table 2—The effect of water-miscible organic solvents on the stability of Est06.
Supplementary Figure 1—Maps of plasmids for cloning and expression of the lipolytic gene est06.
Supplementary Figure 2—Predicted tertiary structure of esterase Est06.
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Dukunde, A., Schneider, D., Lu, M. et al. A novel, versatile family IV carboxylesterase exhibits high stability and activity in a broad pH spectrum. Biotechnol Lett 39, 577–587 (2017). https://doi.org/10.1007/s10529-016-2282-1
- Family IV esterase
- Hormone-sensitive lipase family
- Soil metagenome