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

Verfasser / Beitragende:
[Florence Privé, C Newbold, Naheed Kaderbhai, Susan Girdwood, Olga Golyshina, Peter Golyshin, Nigel Scollan, Sharon Huws]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/13(2015-07-01), 5475-5485
Format:
Artikel (online)
ID: 605506140
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024 7 0 |a 10.1007/s00253-014-6355-6  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-014-6355-6 
245 0 0 |a Isolation and characterization of novel lipases/esterases from a bovine rumen metagenome  |h [Elektronische Daten]  |c [Florence Privé, C Newbold, Naheed Kaderbhai, Susan Girdwood, Olga Golyshina, Peter Golyshin, Nigel Scollan, Sharon Huws] 
520 3 |a 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. 
540 |a The Author(s), 2015 
690 7 |a Rumen  |2 nationallicence 
690 7 |a Lipolysis  |2 nationallicence 
690 7 |a Fatty acid  |2 nationallicence 
690 7 |a Lipase  |2 nationallicence 
690 7 |a Esterase  |2 nationallicence 
690 7 |a Bacteria  |2 nationallicence 
690 7 |a Functional metagenomic  |2 nationallicence 
700 1 |a Privé  |D Florence  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
700 1 |a Newbold  |D C.  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
700 1 |a Kaderbhai  |D Naheed  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
700 1 |a Girdwood  |D Susan  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
700 1 |a Golyshina  |D Olga  |u School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK  |4 aut 
700 1 |a Golyshin  |D Peter  |u School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK  |4 aut 
700 1 |a Scollan  |D Nigel  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
700 1 |a Huws  |D Sharon  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/13(2015-07-01), 5475-5485  |x 0175-7598  |q 99:13<5475  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-014-6355-6  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-014-6355-6  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Privé  |D Florence  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Newbold  |D C.  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kaderbhai  |D Naheed  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Girdwood  |D Susan  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Golyshina  |D Olga  |u School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Golyshin  |D Peter  |u School of Biological Sciences, Bangor University, LL57 2UW, Bangor, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Scollan  |D Nigel  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Huws  |D Sharon  |u Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, SY23 3DA, Aberystwyth, UK  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/13(2015-07-01), 5475-5485  |x 0175-7598  |q 99:13<5475  |1 2015  |2 99  |o 253