Identification and characterization of three novel lipases belonging to families II and V from Anaerovibrio lipolyticus 5ST

Florence Privé, Naheed N Kaderbhai, Susan Girdwood, Hilary J Worgan, Eric Pinloche, Nigel D Scollan, Sharon A Huws, C Jamie Newbold

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3%. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).

Original languageEnglish
Article numbere69076
JournalPLoS ONE
Volume8
Issue number8
DOIs
Publication statusPrint publication - Aug 2013
Externally publishedYes

Fingerprint

Anaerovibrio lipolyticus
Rumen
Lipase
Caprylates
Genes
Laurates
Genome
Bacteria
Lipolysis
Base Composition
Myristic Acid
Ruminants
Enzyme Assays
Enzymes
Substrate Specificity
enzymes
Unsaturated Fatty Acids
biohydrogenation
rumen bacteria
Hydrolysis

Keywords

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Conserved Sequence
  • Enzyme Activation
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Lipase/chemistry
  • Lipolysis
  • Molecular Sequence Data
  • Phylogeny
  • Recombinant Proteins/genetics
  • Sequence Alignment
  • Substrate Specificity
  • Temperature
  • Veillonellaceae/classification

Cite this

Privé, Florence ; Kaderbhai, Naheed N ; Girdwood, Susan ; Worgan, Hilary J ; Pinloche, Eric ; Scollan, Nigel D ; Huws, Sharon A ; Newbold, C Jamie. / Identification and characterization of three novel lipases belonging to families II and V from Anaerovibrio lipolyticus 5ST. In: PLoS ONE. 2013 ; Vol. 8, No. 8.
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abstract = "Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3{\%}. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).",
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Identification and characterization of three novel lipases belonging to families II and V from Anaerovibrio lipolyticus 5ST. / Privé, Florence; Kaderbhai, Naheed N; Girdwood, Susan; Worgan, Hilary J; Pinloche, Eric; Scollan, Nigel D; Huws, Sharon A; Newbold, C Jamie.

In: PLoS ONE, Vol. 8, No. 8, e69076, 08.2013.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification and characterization of three novel lipases belonging to families II and V from Anaerovibrio lipolyticus 5ST

AU - Privé, Florence

AU - Kaderbhai, Naheed N

AU - Girdwood, Susan

AU - Worgan, Hilary J

AU - Pinloche, Eric

AU - Scollan, Nigel D

AU - Huws, Sharon A

AU - Newbold, C Jamie

PY - 2013/8

Y1 - 2013/8

N2 - Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3%. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).

AB - Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3%. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).

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KW - Amino Acid Sequence

KW - Animals

KW - Conserved Sequence

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KW - Gene Expression

KW - Hydrogen-Ion Concentration

KW - Lipase/chemistry

KW - Lipolysis

KW - Molecular Sequence Data

KW - Phylogeny

KW - Recombinant Proteins/genetics

KW - Sequence Alignment

KW - Substrate Specificity

KW - Temperature

KW - Veillonellaceae/classification

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DO - 10.1371/journal.pone.0069076

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JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

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