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Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome

Prive, F. and Newbold, C.J. and Kaderbhai, N.N. and Girdwood, S.G. and Golyshina, O.V. and Golyshin, P.N. and Scollan, N.D. and Huws, S.A. (2015) Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome. Applied Microbiology and Biotechnology, 99 (13). pp. 5474-5485. DOI: 10.1007/s00253-014-6355-6

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Abstract

Improving the health beneficial fatty acid content of meat and milk is a major challenge requiring an increased understanding of rumen lipid metabolism. In this study, we isolated and characterized rumen bacterial lipases/esterases using functional metagenomics. Metagenomic libraries were constructed from DNA extracted from strained rumen fluid (SRF), solid-attached bacteria (SAB) and liquid-associated rumen bacteria (LAB), ligated into a fosmid vector and subsequently transformed into an Escherichia coli host. Fosmid libraries consisted of 7,744; 8,448; and 7,680 clones with an average insert size of 30 to 35 kbp for SRF, SAB and LAB, respectively. Transformants were screened on spirit blue agar plates containing tributyrin for lipase/esterase activity. Five SAB and four LAB clones exhibited lipolytic activity, and no positive clones were found in the SRF library. Fosmids from positive clones were pyrosequenced and twelve putative lipase/esterase genes and two phospholipase genes retrieved. Although the derived proteins clustered into diverse esterase and lipase families, a degree of novelty was seen, with homology ranging from 40 to 78 % following BlastP searches. Isolated lipases/esterases exhibited activity against mostly short- to medium-chain substrates across a range of temperatures and pH. The function of these novel enzymes recovered in ruminal metabolism needs further investigation, alongside their potential industrial uses.

Item Type: Article
Subjects: Research Publications
Departments: College of Natural Sciences > School of Biological Sciences
Date Deposited: 21 Aug 2015 02:47
Last Modified: 23 Sep 2015 02:50
ISSN: 0175-7598
URI: http://e.bangor.ac.uk/id/eprint/5263
Identification Number: DOI: 10.1007/s00253-014-6355-6
Publisher: Springer
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