1. Lecture 10: Bacterial Adaptation Reading assignments in Text: Lengeler et al. 1999 Text: pages 469-483 “Rapid” Enzyme control pages 123-126 ATP / NAD(P)H regulation Lecture 9 Text:pages 674-676 Bacterial diversity pages 700-704 Phylogenetic trees pages 704-716 Early life/ evolution pages 723-728 Food in the real world pages 746-750 Biofilms pages 754-761 Cooperation and methanogens pages 763-774 Bugs in water pages 775-778 Bugs in sediments pages 779-784 Bugs in soil pages 784-792 Bugs in extreme environments pages 879-882 Bugs in food products pages 907-908 Bio-treatment

2. Lecture Overview Bacterial populations (lab conditions) Metabolism GROWTHGROWTH Bacteria as single cells (“cell cycles”) Differentiation Symbiosis Sporulation Bacterial Diversity Adaptation Mechanisms Rapid Slower Change enzyme activities Make new enzymes

3. Rapid adaptation responses/ Demand feeding E. coli M914-C glycerol 14-C 20 amino acids E. coliM9 14-C glycerol Cold Histidine 14-C 19 amino acids Cell growthCell proteins Cold Histidine 14-C MP’s Amino acids Histidine Biosyn. (-) Feed Back inhibition Allows “demand feeding” of Biosynthesis “Allosteric” More active

5. Feedback systems Trp Phe Tyr 1) Sequential FB (-), most logical 2) isozymes, different FB targets 3) cumulative FB(-), most common E. coli M9Rapid growth +Trp Slow, stop growth ? “metabolic imbalance”

7. Global ATP and NADPH allosteric controls E. coli + O 2, rapid growth, [ATP] high N2N2 O2O2 Stops growth, [ATP] stays high CMet. 12 MP’s Biosynthesis ATP NAD(P)H (+/-) ATP (~P) Energy Charge EC = [ATP] + (1/2)[ADP] [ATP]+[ADP]+[AMP] +(1/2)[ADP] Adenylate kinase AMP + ATP 2 ADP Theoretical EC = 0-1.0 Physiological EC = 0.87-0.95, <0.5 XX

9. (Rxns use ATP) ATP(+)  ATP(-)  (Rxns make ATP)  (+)AMP  (-)AMP make use 0.0 1.0  Rxn rates EC Stable response point Rapid and massive use of ATP ATPADP 10 msec Growing 1.0 gram of E. coli LBM9 18.5 mMoles ATP (MW = 507)2.8

10. Allosteric maintainance of reduction power Catabolic Reduction Charge, CRC = [NADH] [NADH] + [NAD+] Low 0.03 - 0.07 Anabolic Reduction Charge, ARC = [NADPH] [NADPH] + [NADP+] High 0.3 - 0.7

12. Covalent modifications of bacterial proteins -Ser-OH -Ser-O- PO 3 -- Kinase ATP Rare -Asp-C-O- PO 3 -- =O -Asp-C-O - =O Kinase-His~P i ATP Common -Glu-C-O - =O Common -Glu-C-O- CH 3 =O SAM~CH 3 -Tyr-OH -Tyr-O- AMP PP i ATP Common AT = Adenyl Trans GS = Glutamine Synthase

14. 280  nm 260   Low NH 3 High NH 3 Glutamine Synthase (GS) protein 12 X (GS) 12 X (GS-Try-AMP) High NH 3 High Enzyme activity Low Enzyme activity

15. Adaptation to high and low ammonium High NH 3 Glu  Keto NADPH (GS-AMP) Gln NH 3 ATP (GS) Gln NH 3 ATP Biosynthesis Low use Trp, His, Carbamyl~P CTP, AMP glucosamine-6-P “met. delegates” ] (-) Cumulative FB inhibition Low NH 3 Glu  Keto NADPH  Keto 2x Glu NADPH No FB inhibition High use GS = “N-pump” Extra pool

17. Sensing N and switching GS The players: GS UT= Uridyl Transferase P AT P-UMP = Adenyl Tranferase GS-AMP NRII = Kinase His~P NRI = Receiver Asp~P GS Enzyme Activity GS mRNA synthesis (-)

19. Sensing N and switching GS NRII (-) Low NH 3 Sensor ? UT [Gln /  Keto] GS P-UMP P AT GS-AMPGS -AMP High activity No FB inhibition NRI ~P DNA GS glnA mRNA More GS protein