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Indian J Microbiol. 2012 Dec;52(4):538-43. doi: 10.1007/s12088-012-0263-1. Epub 2012 Mar 25.

Mining the metagenome of activated biomass of an industrial wastewater treatment plant by a novel method.

Author information

1
Environmental Genomics Division, National Environmental Engineering Research Institute, CSIR, Nehru Marg, Nagpur, 440020 Maharashtra India.

Abstract

Metagenomic libraries herald the era of magnifying the microbial world, tapping into the vast metabolic potential of uncultivated microbes, and enhancing the rate of discovery of novel genes and pathways. In this paper, we describe a method that facilitates the extraction of metagenomic DNA from activated sludge of an industrial wastewater treatment plant and its use in mining the metagenome via library construction. The efficiency of this method was demonstrated by the large representation of the bacterial genome in the constructed metagenomic libraries and by the functional clones obtained. The BAC library represented 95.6 times the bacterial genome, while, the pUC library represented 41.7 times the bacterial genome. Twelve clones in the BAC library demonstrated lipolytic activity, while four clones demonstrated dioxygenase activity. Four clones in pUC library tested positive for cellulase activity. This method, using FTA cards, not only can be used for library construction, but can also store the metagenome at room temperature.

KEYWORDS:

Activated biomass; Cellulase; FTA elute micro card; Lipase; Metagenomic library

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