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«Physical chemistry of impact of metal ions on the metabolism 
of bacterium Azospirillum brasilense»

PhD dissertation of Anatoly Chernyshev, saratov State University, May 1999

Table of Contents

Section

 Page
List of abridgements and symbols 4
Introduction 5
Ch. 1. Literature review 10
1.1. Role of metal ions in physiology of a bacterial cell 10
1.1.1. Passive diffuzion 12
1.1.2. Facilitated diffusion 25
1.1.3. Active transport 30
1.1.4. Adsorption and formation of a new phase 32
1.2. Biological catalysis onto binuclear metal complexes 48
Ch. 2 Materials and methods 72
2.1. Main object (A. brasilense Sp 245) and its cultivation 72
2.2. Isolation and purification of glutamine synthetase 72
2.3. Determination of activity and the adenylylation state of glutamine synthetase 74
2.4. Determination of protein concentration 74

2.5. Determination of the activity of glutamine synthetase in the presence of metal ions

 75
2.6. Light microscopy 76
2.7. Isolation of mineral crystals 76
2.8. Structure investigations of mineral crystals 76
2.8.1. IR-spectroscopy 76
2.8.2. X-ray phase analysis 77
2.8.3. Thermogravimetric and differential thermic analysis 77
2.8.4. Ionic chromatography and element analysis 77
2.9. Atom emission and atom absorption spectrometry 77
2.10. Sampling of dry biomass of A. brasilense 77
2.11. Fourier IR spectroscopy 78
2.12. Spectroturbidimetry 78
2.13. Phosphatase activity of lanthanides 78
2.14. Glutamine synthetase activity in the presence of chalcogenopirilium salts and 1,5-diketones 79
2.15. Electrochemical study of kinetic properties of glutamine synthetase and ATP hydrolysis in the presence of lanthanide ions 79
Ch. 3. Results and discussion 80
3.1. Formation of struvite crystals during cultivation of A. brasilense 80
3.2. Absorption of metal ions by A. brasilense cells 89
3.3. Interaction of A. brasilense glutamine synthetase with ions of s-, p- and d-elements 98
3.4. A. brasilense glutamine synthetase behaviour in transferase reaction with lanthanide ions and intrinsic phosphatase activity of lanthanide ions 114
3.5. Interaction of A. brasilense glutamine synthetase with elementoorganic ions  128
Afterword 140
Conclusions 144
Literature cited (342 sources) 146
Acknowledgements 174
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