Physiology of B acteria . Growth and reproduction of Bacteria

1. Chair of Microbiology, Virology, and Immunology Physiology of Bacteria.Growth and reproduction of Bacteria

2. Lecture schedule 1. Chemical composition of Bacteria 2. Cell metabolism 3. Contstructive metabolism метаболізм 4. Types of microbial nutrition 5. Bacterial transport systems 6. Types of respiration 7 Growth and reproduction of microbes 8. Nutrient media

3. Metabolism refers to all the biochemical reactions that occur in a cell or organism. The study of bacterial metabolism focuses on the chemical diversity of substrate oxidations and dissimilation reactions (reactions by which substrate molecules are broken down), which normally function in bacteria to generate energy.

4. Chemical composition of bacteria

5. Bacterial cell consists of: Water – 70-90 % Dry weight – 10-30 % Proteins – 55 %, 2,35 million of molecules, 1850 different types of molecules RNA – 20,5 %, 250000 molecules, 660 different types of molecules DNA – 3,1 %, 2 molecules Lipids – 9 %, 22 million of molecules Lipopolysaccharides –3,4 %, 1,5 million of molecules Peptidoglycan – 1 molecule

6. Microbial metabolism 1. Catabolism (Dissimilation) - Pathways that breakdown organic substrates (carbohydrates, lipids, & proteins) to yield metabolic energy for growth and maintenance. 2. Anabolism (Assimilation) - Assimilatory pathways for the formation of key intermediates and then to end products (cellular components). 4. Intermediary metabolism-Integrate two processes

7. Catabolism Substrate-level phosphorylation Fermentation Glycolysis (EMP pathway) Aerobic respiration Pyruvate: universal intermediate

8. The bacterial cell is a highly specialized energy transformer. Chemical energy generated by substrate oxidations is conserved by formation of high-energy compounds such as adenosine diphosphate (ADP) and adenosine triphosphate (ATP) or compounds containing the thioester bond O ║ (R –C ~ S – R), such as acetyl ~ S-coenzyme A

9. Another form of energy - transmembrane potential - ΔμН+

10. Chemiosmosis • Production of ATP in Electron Transport • Electrochemical Gradient Formed between membranes • H+ (Protons) generated from NADH • Electrical Force (+) & pH Force (Acid) • Gradient formed • ATPase enzyme that channels H+ from High to Low concentration • 3 ATP/NADH • 2 ATP/NADH

11. Sources of metabolic energy Respiration: chemical reduction of an electron acceptor through a specific series of electron carriers in the membrane. The electron acceptor is commonly O2, but CO2, SO42-, and NO3- are employed by some microorganisms. Photosynthesis: similar to respiration except that the reductant and oxidant are created by light energy. Respiration can provide photosynthetic organisms with energy in the absence of light. Substrate-level phosphorylation Fermentation: metabolic process in which the final electron acceptor is an organic compound.

12. The Krebs cycle intermediate compounds serve as precursor molecules (building blocks) for the energy-requiring biosynthesis of complex organic compounds in bacteria. Degradation reactions that simultaneously produce energy and generate precursor molecules for the biosynthesis of new cellular constituents are called amphibolic.

13. Energy Requirements Oxidation of organic compounds- Chemotrophs Sunlight- Phototrophs

14. Metabolic Requirements Carbon source - Autotrophs (lithotrophs): use CO2 as the C source Photosynthetic autotrophs: use light energy Chemolithotrophs: use inorganics - Heterotrophs (organotrophs): use organic carbon (eg. glucose) for growth. - Clinical Labs classify bacteria by the carbon sources (eg. Lactose) & the end products (eg. Ethanol,…). Nitrogen source Ammonium (NH4+) is used as the sole N source by most microorganisms. Ammonium could be produced from N2 by nitrogen fixation, or from reduction of nitrate (NO3-)andnitrite (NO2).

15. Physiologic types of bacterial existence Energy Source Oxidation of organic compounds- Chemotrophs Sunlight- Phototrophs Carbon Source Organic- Heterotrophs Inorganic- Autotrophs Electrone donor Inorganic- Lithotrophs Оrganic-Organotrophs Chemoorganoheterotrophic bacteria

16. Metabolic Requirements Sulfur source A component of several coenzymes and amino acids. Most microorganisms can use sulfate (SO42-) as the S source. Phosphorus source - A component of ATP, nucleic acids, coenzymes, phospholipids, teichoic acid, capsular polysaccharides; also is required for signal transduction. - Phosphate (PO43-) is usually used as the P source.

17. Mineral source • - Required for enzyme function. • For most microorganisms, it is necessary to provide sources • of K+, Mg2+, Ca2+, Fe2+, Na+ and Cl-. • Many other minerals (eg., Mn2+, Mo2+, Co2+, Cu2+ and Zn2+) • can be provided in tap water or as contaminants of other • medium ingredients. • Uptake of Fe is facilitated by production of siderophores • (Iron-chelating compound, eg. Enterobactin). • Growth factors: organic compounds (e.g., amino acids, sugars, nucleotides, vitamines) a cell must contain in order to grow but which it is unable to synthesize. Purines and pyrimidines: required for synthesis of nucleic acids (DNA and RNA); • Amino acids: required for the synthesis of proteins; • Vitamins: needed as coenzymes and functional groups of certain enzymes.

18. Transport systems The proteins that mediate the passage of solutes through membranes are referred to as transport systems, carrier proteins, porters, and permeases. Transport systems operate by one of three transport processes. In a uniport process, a solute passes through the membrane unidirectionally. In symport processes (cotransport) two solutes must be transported in the same direction at the same time; in antiport processes (exchange diffusion), one solute is transported in one direction simultaneously as a second solute is transported in the opposite direction.

19. Transport systems

20. Diffusion systems • passive diffusion • facilitated diffusion • ion-driven transport • binding protein dependent transport • group translocation

21. Membrane is selectively permeable • Few molecules pass through freely • Movement involves both active and passive processes

22. Passive processes • no energy (ATP) required • Along gradient • simple diffusion, facilitated diffusion, osmosis

23. Simple diffusion • Facilitated diffusion • Can reduce concentration gradient but can’t create one

24. Osmosis • Osmotic pressure

25. Active processes • energy (ATP) required • Active transport • Group translocation

26. Facilitated diffusion

27. Active transport

28. Transport systems

29. TEMPERATURE • One of the most important factors • optimal growth temperature • temperature range at which the highest rate of reproduction occurs • optimal growth temperature for human pathogens ????

30. TEMPERATURE • Microorganisms can be categorized based on their optimal temperature requirements • Psychrophiles • 0 - 20 ºC • Mesophiles • 20 - 40 ºC • Thermophiles • 40 - 90 ºC • Most bacteria are mesophiles especially pathogens that require 37 ºC

31. BACTERIAL TEMPERATURE REQUIREMENTS 100 Psychrophile Thermophile % Max Growth 50 Mesophile 0 0 0C 37 0C 90 0C Variable

32. Effects of Temperature on Growth

33. Thermophiles 70o-110o Mesophiles 10o-50o For lecture only BC Yang

34. TEMPERATURE • Psychrophiles • some will exist below 0 oC if liquid water is available • oceans • refrigerators • freezers Pigmented bacteria in Antarctic ice

35. TEMPERATURE • Mesophiles • most human flora and pathogens

36. TEMPERATURE • Thermophiles • hot springs • effluents from laundromat • deep ocean thermal vents

37. Respiration in Bacteria Obligate Aerobe Microaerophile Obligate Anaerobe Facultative Anaerobe (Facultative Aerobe) Aerotolerant Anaerobe Capneic bacteria

38. Categories of Oxygen Requirement Aerobe – utilizes oxygen and can detoxify it obligate aerobe - cannot grow without oxygen (Mycobacterium tuberculosis, Micrococcus spp., Bacillus spp., Pseudomonas spp. facultative anaerobe – utilizes oxygen but can also grow in its absence (Echericihia spp., Salmonella spp., Sta[phylococcus spp.) microaerophylic – requires only a small amount of oxygen (Helycobacter spp., Lactobacillus spp.) 38

39. Categories of Oxygen Requirement Anaerobe – does not utilize oxygen obligate anaerobe - lacks the enzymes to detoxify oxygen so cannot survive in an oxygen environment (Clostridium spp., Bacteroides spp.) aerotolerance anaerobes – do no utilize oxygen but can survive and grow in its presence (Streptococcus pyogenes) 39

40. Carbon Dioxide Requirement All microbes require some carbon dioxide in their metabolism. capneic – grows best at higher CO2 tensions than normally present in the atmosphere (Brucella abortus) 40

41. OXYGEN Obligate Aerobe Facultative Anaerobe Obligate Anaerobe

42. Four Toxic Forms of Oxygen

43. Four Toxic Forms of Oxygen

44. Enzymes and Their Role in Metabolism Enzymes, organic catalysts of a highly molecular structure, are produced by the living cell. They are of a protein nature, are strictly specific in action, and play an important part in the metabolism of micro-organisms. Their specificity is associated with active centres formed by a group of amino acids.

45. Some enzymes are excreted by the cell into the environment (exoenzymes) for breaking down complex colloid nutrient materials while other enzymes are contained inside the cell (endoenzymes).

46. Bacterial enzymes are subdivided into some groups: 1. Hydrolases which catalyse the breakdown of the link between the carbon and nitrogen atoms, between the oxygen and sulphur atoms, binding one molecule of water (esterases. glucosidases, proteases. amilases, nucleases, etc.). 2. Transferases perform catalysis by transferring certain radicals from one molecule to another (transglucosidases, transacylases. transaminases). 3. Oxidative enzymes (oxyreductases) which catalyse the oxidation-reduction processes (oxidases, dehydrogenases, peroxidases, catalases). 4. Isomerases and racemases play an important part in carbohydrate metabolism. Rearrangement atoms of a molecule. 5. Lyases (remove chemical groups from molecules without adding water). 6. Lygases (join two molecules together and usually require energy from ATP).

47. Enzymes

48. Significance of the enzymes With the help of amylase produced by mould fungi starch is saccharified and this is employed in beer making, industrial alcohol production and bread making. Proteinases produced by microbes are used for removing the hair from hides, tanning hides, liquefying the gelatinous layer from films during regeneration, and for dry cleaning. Fibrinolysin produced by streptococci dissolves the thrombi in human blood vessels. Enzymes which hydrolyse cellulose aid in an easier assimilation of rough fodder.

49. Due to the application of microbial enzymes, the medical industry has been able to obtain alkaloids, polysaccharides, and steroids (hydrocortisone, prednisone, prednisolone. etc.). Bacteria play an important role in the treatment of caouichouc, coffee, cocoa, and tobacco. Enzymes permit some species of microorganisms to assimilate methane. butane, and other hydrocarbons, and to synthesize complex organic compounds from them. With the help of the enzymatic ability of yeasts in special-type industrial installations protein-vitamin concentrates (PVC) can be obtained from waste products of petroleum (paraffin’s).

50. Metabolism Results in Reproduction Microbial growth – an increase in a population of microbes rather than an increase in size of an individual Result of microbial growth is discrete colony – an aggregation of cells arising from single parent cell Reproduction results in growth