Abstract
Glutamate dehydrogenase (L-glutamate:NADP+ oxidoreductase [deaminating], EC 1.4.1.4) has been purified from E. coli B/r. The purity of the enzyme preparation has been established by polyacrylamide gel electrophoresis, ultracentrifugation, and gel filtration. A molecular weight of 300,000 ± 20,000 has been calculated for the enzyme from sedimentation equilibrium measurements. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate and sedimentation equilibrium measurements in guanidine hydrochloride have revealed that glutamate dehydrogenase consists of polypeptide chains with the identical molecular weight of 50,000 ± 5,000. The results of molecular weight determination lead to the proposal that glutamate dehydrogenase is a hexamer of subunits with identical molecular weight. The stability and kinetics of purified glutamate dehydrogenase were also studied. The enzyme remains active when heat treated or when left at room temperature for several months but it is inactivated by freezing. The Michaelis constants of glutamate dehydrogenase are 1,100, 640, and 40 μM for ammonia, 2 oxoglutarate, and reduced nicotinamide adenine dinucleotide phosphate, respectively.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 775-783 |
| Number of pages | 9 |
| Journal | Journal of Bacteriology |
| Volume | 124 |
| Issue number | 2 |
| State | Published - 1975 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Immunology
Cite this
Glutamate dehydrogenase from Escherichia coli : purification and properties. / Sakamoto, N.; Kotre, A. M.; Savageau, M. A.
In: Journal of Bacteriology, Vol. 124, No. 2, 1975, p. 775-783.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Glutamate dehydrogenase from Escherichia coli
T2 - purification and properties
AU - Sakamoto, N.
AU - Kotre, A. M.
AU - Savageau, M. A.
PY - 1975
Y1 - 1975
N2 - Glutamate dehydrogenase (L-glutamate:NADP+ oxidoreductase [deaminating], EC 1.4.1.4) has been purified from E. coli B/r. The purity of the enzyme preparation has been established by polyacrylamide gel electrophoresis, ultracentrifugation, and gel filtration. A molecular weight of 300,000 ± 20,000 has been calculated for the enzyme from sedimentation equilibrium measurements. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate and sedimentation equilibrium measurements in guanidine hydrochloride have revealed that glutamate dehydrogenase consists of polypeptide chains with the identical molecular weight of 50,000 ± 5,000. The results of molecular weight determination lead to the proposal that glutamate dehydrogenase is a hexamer of subunits with identical molecular weight. The stability and kinetics of purified glutamate dehydrogenase were also studied. The enzyme remains active when heat treated or when left at room temperature for several months but it is inactivated by freezing. The Michaelis constants of glutamate dehydrogenase are 1,100, 640, and 40 μM for ammonia, 2 oxoglutarate, and reduced nicotinamide adenine dinucleotide phosphate, respectively.
AB - Glutamate dehydrogenase (L-glutamate:NADP+ oxidoreductase [deaminating], EC 1.4.1.4) has been purified from E. coli B/r. The purity of the enzyme preparation has been established by polyacrylamide gel electrophoresis, ultracentrifugation, and gel filtration. A molecular weight of 300,000 ± 20,000 has been calculated for the enzyme from sedimentation equilibrium measurements. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate and sedimentation equilibrium measurements in guanidine hydrochloride have revealed that glutamate dehydrogenase consists of polypeptide chains with the identical molecular weight of 50,000 ± 5,000. The results of molecular weight determination lead to the proposal that glutamate dehydrogenase is a hexamer of subunits with identical molecular weight. The stability and kinetics of purified glutamate dehydrogenase were also studied. The enzyme remains active when heat treated or when left at room temperature for several months but it is inactivated by freezing. The Michaelis constants of glutamate dehydrogenase are 1,100, 640, and 40 μM for ammonia, 2 oxoglutarate, and reduced nicotinamide adenine dinucleotide phosphate, respectively.
UR - http://www.scopus.com/inward/record.url?scp=0016697801&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0016697801&partnerID=8YFLogxK
M3 - Article
C2 - 241744
AN - SCOPUS:0016697801
VL - 124
SP - 775
EP - 783
JO - Journal of Bacteriology
JF - Journal of Bacteriology
SN - 0021-9193
IS - 2
ER -