caa a22 4500 605506140 CHVBK 20210128100625.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s00253-014-6355-6 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6355-6 Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome [Elektronische Daten] [Florence Privé, C Newbold, Naheed Kaderbhai, Susan Girdwood, Olga Golyshina, Peter Golyshin, Nigel Scollan, Sharon Huws] 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 35kbp 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. The Author(s), 2015 Rumen nationallicence Lipolysis nationallicence Fatty acid nationallicence Lipase nationallicence Esterase nationallicence Bacteria nationallicence Functional metagenomic nationallicence Privé Florence Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Newbold C. Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Kaderbhai Naheed Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Girdwood Susan Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Golyshina Olga School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK aut Golyshin Peter School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK aut Scollan Nigel Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Huws Sharon Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5475-5485 0175-7598 99:13<5475 2015 99 253 https://doi.org/10.1007/s00253-014-6355-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6355-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Privé Florence Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 700 1- Newbold C. Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 700 1- Kaderbhai Naheed Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 700 1- Girdwood Susan Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 700 1- Golyshina Olga School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK aut NATIONALLICENCE 700 1- Golyshin Peter School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK aut NATIONALLICENCE 700 1- Scollan Nigel Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 700 1- Huws Sharon Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/13(2015-07-01), 5475-5485 0175-7598 99:13<5475 2015 99 253