Purification and properties of glutamine synthetase from Bacillus polymyxa |
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Authors: | Mukti Ojha Salomé Kantengwa |
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Institution: | (1) Laboratory of Microbiology, Department of Plant Biology, University of Geneva, 3 Place de l'Université, CH-1211 Geneva 4, Switzerland |
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Abstract: | Glutamine synthetase (EC 6.3.1.2) was purified to homogeneity from a free-living nitrogen fixing bacteria, Bacillus polymyxa. The holoenzyme, relative molecular mass (Mr) of 600 000 is composed of monomeric sub-units of 60 000 (Mr). The isoelectric point of the sub-units was 5.2. The pH optimum for the biosynthetic and transferase enzyme activity was 8.2 and 7.8, respectively. The apparent K
m values (K
m
app
) in the biosynthetic reaction for glutamate, NH4Cl and ATP were 3.2, 0.22 and 1 mM, respectively. In the transferase reaction the K
m values for glutamine, hydroxylamine and ADP were 6.5, 3.5 and 8×10-4 mM respectively. L-Methionine-D-L-sulfoximine was a very potent inhibitor in both biosynthetic and transferase reactions. Similar to most Gram positive bacteria there was no evidence of in vivo adenylylation and the enzyme seemed to be mainly regulated by feed-back mechanism.Abbreviations PMSF
phenylmethylsulfonylfluoride
- TCA
trichloroacetic acid
- GS
glutamine synthetase
- MSO
L-Methionine-D-L-sulfoximine
- SDS-PAGE
sodium dodecyl sulfatepolyacrylamide gel electrophoresis
- SVPDE
snake venum phosphodiesterase |
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Keywords: | Bacillus polymyxa Glutamine synthetase Properties |
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