An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum

C James Newbold; Neil R McEwan; Roger E Calza; Emilie N Chareyron; Stéphane M Duval; Sylvain C P Eschenlauer; Freda M McIntosh; Nancy Nelson; Anthony J Travis; R John Wallace

doi:10.1016/j.femsle.2005.04.034

An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum

C James Newbold, Neil R McEwan, Roger E Calza, Emilie N Chareyron, Stéphane M Duval, Sylvain C P Eschenlauer, Freda M McIntosh, Nancy Nelson, Anthony J Travis, R John Wallace

Research output: Contribution to journal › Article › peer-review

15 Citations (Scopus)

Abstract

An NAD(+)-dependent glutamate dehydrogenase (GDH; EC 1.4.1.24) was cloned from the ruminal ciliate protozoan, Entodinium caudatum. The gene had high sequence similarity to GDH genes from the Bacteroides (class)--a class of bacteria which is highly represented in the rumen. When expressed in Escherichia coli the enzyme had a high affinity for ammonia and alpha-ketoglutarate (apparent K(m) of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent K(m) of 98 mM). GDH activity and GDH mRNA concentration were increased by incubating washed E. caudatum cells with ammonia and antibiotics. These results suggest that the GDH is an anabolic enzyme catalysing the assimilation of ammonia by E. caudatum in the rumen and that the gene was probably acquired by lateral gene transfer from a ruminal bacterium.

Original language	English
Pages (from-to)	113-21
Number of pages	9
Journal	FEMS Microbiology Letters
Volume	247
Issue number	2
DOIs	https://doi.org/10.1016/j.femsle.2005.04.034
Publication status	Print publication - 15 Jun 2005
Externally published	Yes

Keywords

Amino Acid Sequence
Ammonia/metabolism
Animals
Bacteroides/genetics
Base Sequence
Ciliophora/enzymology
Cloning, Molecular
DNA, Protozoan/chemistry
Escherichia coli/genetics
Gene Expression Regulation
Gene Transfer, Horizontal
Genes, Protozoan
Glutamate Dehydrogenase/genetics
Glutamic Acid/metabolism
Ketoglutaric Acids/metabolism
Molecular Sequence Data
Phylogeny
RNA, Messenger/analysis
RNA, Protozoan/analysis
Recombinant Proteins/metabolism
Sequence Homology, Amino Acid
Substrate Specificity

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10.1016/j.femsle.2005.04.034

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title = "An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum",

abstract = "An NAD(+)-dependent glutamate dehydrogenase (GDH; EC 1.4.1.24) was cloned from the ruminal ciliate protozoan, Entodinium caudatum. The gene had high sequence similarity to GDH genes from the Bacteroides (class)--a class of bacteria which is highly represented in the rumen. When expressed in Escherichia coli the enzyme had a high affinity for ammonia and alpha-ketoglutarate (apparent K(m) of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent K(m) of 98 mM). GDH activity and GDH mRNA concentration were increased by incubating washed E. caudatum cells with ammonia and antibiotics. These results suggest that the GDH is an anabolic enzyme catalysing the assimilation of ammonia by E. caudatum in the rumen and that the gene was probably acquired by lateral gene transfer from a ruminal bacterium.",

keywords = "Amino Acid Sequence, Ammonia/metabolism, Animals, Bacteroides/genetics, Base Sequence, Ciliophora/enzymology, Cloning, Molecular, DNA, Protozoan/chemistry, Escherichia coli/genetics, Gene Expression Regulation, Gene Transfer, Horizontal, Genes, Protozoan, Glutamate Dehydrogenase/genetics, Glutamic Acid/metabolism, Ketoglutaric Acids/metabolism, Molecular Sequence Data, Phylogeny, RNA, Messenger/analysis, RNA, Protozoan/analysis, Recombinant Proteins/metabolism, Sequence Homology, Amino Acid, Substrate Specificity",

author = "Newbold, {C James} and McEwan, {Neil R} and Calza, {Roger E} and Chareyron, {Emilie N} and Duval, {St{\'e}phane M} and Eschenlauer, {Sylvain C P} and McIntosh, {Freda M} and Nancy Nelson and Travis, {Anthony J} and Wallace, {R John}",

year = "2005",

month = jun,

day = "15",

doi = "10.1016/j.femsle.2005.04.034",

language = "English",

volume = "247",

pages = "113--21",

journal = "FEMS Microbiology Letters",

issn = "0378-1097",

publisher = "Oxford University Press",

number = "2",

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TY - JOUR

T1 - An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum

AU - Newbold, C James

AU - McEwan, Neil R

AU - Calza, Roger E

AU - Chareyron, Emilie N

AU - Duval, Stéphane M

AU - Eschenlauer, Sylvain C P

AU - McIntosh, Freda M

AU - Nelson, Nancy

AU - Travis, Anthony J

AU - Wallace, R John

PY - 2005/6/15

Y1 - 2005/6/15

N2 - An NAD(+)-dependent glutamate dehydrogenase (GDH; EC 1.4.1.24) was cloned from the ruminal ciliate protozoan, Entodinium caudatum. The gene had high sequence similarity to GDH genes from the Bacteroides (class)--a class of bacteria which is highly represented in the rumen. When expressed in Escherichia coli the enzyme had a high affinity for ammonia and alpha-ketoglutarate (apparent K(m) of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent K(m) of 98 mM). GDH activity and GDH mRNA concentration were increased by incubating washed E. caudatum cells with ammonia and antibiotics. These results suggest that the GDH is an anabolic enzyme catalysing the assimilation of ammonia by E. caudatum in the rumen and that the gene was probably acquired by lateral gene transfer from a ruminal bacterium.

AB - An NAD(+)-dependent glutamate dehydrogenase (GDH; EC 1.4.1.24) was cloned from the ruminal ciliate protozoan, Entodinium caudatum. The gene had high sequence similarity to GDH genes from the Bacteroides (class)--a class of bacteria which is highly represented in the rumen. When expressed in Escherichia coli the enzyme had a high affinity for ammonia and alpha-ketoglutarate (apparent K(m) of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent K(m) of 98 mM). GDH activity and GDH mRNA concentration were increased by incubating washed E. caudatum cells with ammonia and antibiotics. These results suggest that the GDH is an anabolic enzyme catalysing the assimilation of ammonia by E. caudatum in the rumen and that the gene was probably acquired by lateral gene transfer from a ruminal bacterium.

KW - Amino Acid Sequence

KW - Ammonia/metabolism

KW - Animals

KW - Bacteroides/genetics

KW - Base Sequence

KW - Ciliophora/enzymology

KW - Cloning, Molecular

KW - DNA, Protozoan/chemistry

KW - Escherichia coli/genetics

KW - Gene Expression Regulation

KW - Gene Transfer, Horizontal

KW - Genes, Protozoan

KW - Glutamate Dehydrogenase/genetics

KW - Glutamic Acid/metabolism

KW - Ketoglutaric Acids/metabolism

KW - Molecular Sequence Data

KW - Phylogeny

KW - RNA, Messenger/analysis

KW - RNA, Protozoan/analysis

KW - Recombinant Proteins/metabolism

KW - Sequence Homology, Amino Acid

KW - Substrate Specificity

U2 - 10.1016/j.femsle.2005.04.034

DO - 10.1016/j.femsle.2005.04.034

M3 - Article

C2 - 15921862

SN - 0378-1097

VL - 247

SP - 113

EP - 121

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

IS - 2

ER -

An NAD(+)-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum

Abstract

Keywords

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