Mining lipolytic enzymes in community DNA from high Andean soils using a targeted approach

Antonie van Leeuwenhoek
Daniel Borda-MolinaSandra Baena

Abstract

Microbial enrichments cultures are a useful strategy to speed up the search for enzymes that can be employed in industrial processes. Lipases have gained special attention because they show unique properties such as: broad substrate specificity, enantio- and regio-selectivity and stability in organic solvents. A major goal is to identify novel lipolytic enzymes from microorganisms living in cold extreme environments such as high Andean soils, of relevance to our study being their capability be used in industrial processes. Paramo and glacier soils from the Nevados National Park in Colombia were sampled and microbial communities enriched through a fed-batch fermentation using olive oil as an inductor substrate. After 15 days of enrichment under aerobic conditions, total DNA was extracted. Subsequently, metagenomic libraries were constructed in the cosmid vector pWEB-TNC™. After functional screening, twenty and eighteen lipolytic clones were obtained from Paramo and Glacier soil enrichments, respectively. Based on lipid hydrolysis halo dimensions, the clone (Gla1) from a glacier enrichment was selected. A gene related to lipolytic activity was subcloned to evaluate enzyme properties. Phylogenetic analysis of the identified gene s...Continue Reading

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Methods Mentioned

BETA
electrophoresis
X-ray

Software Mentioned

Autodock Vina
Mollegro
MetaMQAPII
PyMOL
ProtParam
SignalP
TASSER
MetaGeneMark
[UNK] 1000
Swiss

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