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

Department of Life Sciences

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 α-ketoglutarate (apparent Km of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent Km 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)	113-121
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	Published - 01 Jun 2005

Keywords

Ammonia uptake
Entodinium caudatum
Glutamate dehydrogenase
Protozoa
Rumen

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10.1016/j.femsle.2005.04.034

http://hdl.handle.net/2160/3094

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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 α-ketoglutarate (apparent Km of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent Km 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 = "Ammonia uptake, Entodinium caudatum, Glutamate dehydrogenase, Protozoa, Rumen",

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}",

note = "Newbold, C. J., McEwan, N. R., Calza, R. E., Chareyron, E. N., Duval, S. M., Eschenlauer, S. C .P., McIntosh, F. M., Nelson, N., Travis, A. J., Wallace, R. J. (2005). An NAD+-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum. FEMS Microbiology Letters, 247 (2), 113-121 Keywords: Ammonia uptake; Entodinium caudatum; Glutamate dehydrogenase; Protozoa; Rumen",

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

language = "English",

volume = "247",

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journal = "FEMS Microbiology Letters",

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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

N1 - Newbold, C. J., McEwan, N. R., Calza, R. E., Chareyron, E. N., Duval, S. M., Eschenlauer, S. C .P., McIntosh, F. M., Nelson, N., Travis, A. J., Wallace, R. J. (2005). An NAD+-dependent glutamate dehydrogenase cloned from the ruminal ciliate protozoan, Entodinium caudatum. FEMS Microbiology Letters, 247 (2), 113-121 Keywords: Ammonia uptake; Entodinium caudatum; Glutamate dehydrogenase; Protozoa; Rumen

PY - 2005/6/1

Y1 - 2005/6/1

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 α-ketoglutarate (apparent Km of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent Km 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 α-ketoglutarate (apparent Km of 2.33 and 0.71 mM, respectively) and a low affinity for glutamate (apparent Km 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 - Ammonia uptake

KW - Entodinium caudatum

KW - Glutamate dehydrogenase

KW - Protozoa

KW - Rumen

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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

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Keywords

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