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Chapter 5

Chapter 5. Microbial Nutrition. The Common Nutrient Requirements. Macroelements (macronutrients) C, O, H, N, S, P, K, Ca, Mg, and Fe required in relatively large amounts Micronutrients ( trace elements ) Mn, Zn, Co, Mo, Ni, and Cu required in trace amounts

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Chapter 5

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  1. Chapter 5 Microbial Nutrition

  2. The Common Nutrient Requirements • Macroelements (macronutrients) • C, O, H, N, S, P, K, Ca, Mg, and Fe • required in relatively large amounts • Micronutrients (trace elements) • Mn, Zn, Co, Mo, Ni, and Cu • required in trace amounts • often supplied in water or in media components

  3. Autotroph and Heterotroph • All organisms require Carbon, Hydrogen, and Oxygen. Carbon is needed for the backbone of all organic molecules. • In addition all organisms require a source of electrons. Electrons are involved in oxidation-reduction reactions in the cell, electron transport chains, and pumps that drive molecules against a concentration gradient on cell membranes. • Organic molecules that supply, carbon, hydrogen, and oxygen are reduced and donate electrons for biosynthesis.

  4. Requirements for Carbon, Hydrogen, and Oxygen • often satisfied together • carbon source often provides H, O and electrons • autotrophs • use carbon dioxide as their sole or principal carbon source • heterotrophs • use organic molecules as carbon sources

  5. Autotrophs • CO2 is used by many microorganisms as the source of Carbon. • Autotrophs have the capacity to reduce it , to form organic molecules. • Photosynthetic bacteria are Photoautotrophs that are able to fix CO2 and use light as their energy source.

  6. Heterotrophs • Organisms that use organic molecules as their source of carbon are Heterotrophs. The most common heterotrophs use organic compounds for both energy and their source of carbon. • Microorganisms are versatile in their ability to use diverse sources of carbon. Burkholderia cepacia can use over 100 different carbon compounds. • Methylotrophic bacteria utilize methanol, methane, and formic acid.

  7. Comparison of Nutritional Modes Study table adapted from Microbiology, Prescott, Chapter Five.

  8. Photolithotrophic autotrophs • Use light energy and have CO2 as their carbon source. • Cyanobacteria uses water as the electron donor and release oxygen • Purple and green sulfur bacteria use inorganic donors like hydrogen and hydrogen sulfide for electrons

  9. Chemoorganotrophic heterotrophs • Use organic compounds as sources of energy,hydrogen, electrons and carbon • Pathogenic organisms fall under this category of nutrition

  10. Photoorganoheterotrophs • Common inhabitants of polluted streams. These bacteria use organic matter as their electron donor and carbon source. • They use light as their source of energy • Important ecological organisms

  11. Chemolithotrophic autotrophs • Autotrophs • Oxidize reduce inorganic compounds such as iron, nitrogen, or sulfur molecules • Derive energy and electrons for biosynthesis • Carbon dioxide is the carbon source

  12. Requirements for Nitrogen • Nitrogen is required for the synthesis of amino acids that compose the structure of proteins, purines and pyrimidines the bases of both DNA and RNA, and for other derivative molecules such as glucosamine. • Many microorganisms can use the nitrogen directly from amino acids. The amino group ( NH2) is derived from ammonia through the action of enzymes such as glutamate dehydrogenase. • Most photoautotrophs and many nonphotosynthetic microorganisms reduce nitrate to ammonia and assimilate nitrogen through nitrate reduction. A variety of bacteria are involved in the nitrogen cycle such as Rhizobium which is able to use atmospheric nitrogen and convert it to ammonia. ( Found on the roots of legumes like soy beans and clover) These compounds are vital for the Nitrogen cycle and the incorporation of nitrogen into plants to make nitrogen comounds.

  13. Phosphorous • Phosphorous is present in phospholipids( membranes), Nucleic acids( DNA and RNA), coenzymes, ATP, some proteins, and other key cellular components. • Inorganic phosphorous is derived from the environment in the form of phosphates. Some microbes such as E. coli can use organophosphates such as hexose – 6-phosphates .

  14. Mixotrophy • Chemical energy – source organic • Inorganic H/e- donor • Organic carbon source

  15. Requirements for Nitrogen, Phosphorus, and Sulfur • Needed for synthesis of important molecules (e.g., amino acids, nucleic acids) • Nitrogen supplied in numerous ways • Phosphorus usually supplied as inorganic phosphate • Sulfur usually supplied as sulfate via assimilatory sulfate reduction

  16. Sources of nitrogen • organic molecules • ammonia • nitrate via assimilatory nitrate reduction • nitrogen gas via nitrogen fixation

  17. Growth Factors • organic compounds • essential cell components (or their precursors) that the cell cannot synthesize • must be supplied by environment if cell is to survive and reproduce

  18. Classes of growth factors • amino acids • needed for protein synthesis • purines and pyrimidines • needed for nucleic acid synthesis • vitamins • function as enzyme cofactors

  19. Amino acids Proteins

  20. Bases of nucleic acids • Adenine and guanine are purines • Cytosine, thymine, and uracil are pyrimidines • Also found in energy triphosphates( ATP and GTP)

  21. Practical importance of growth factors • development of quantitative growth-response assays for measuring concentrations of growth factors in a preparation • industrial production of growth factors by microorganisms

  22. Uptake of Nutrients by the Cell • Some nutrients enter by passive diffusion • Most nutrients enter by: • facilitated diffusion • active transport • group translocation

  23. Passive Diffusion • molecules move from region of higher concentration to one of lower concentration because of random thermal agitation • H2O, O2 and CO2 often move across membranes this way

  24. Facilitated Diffusion • Similar to passive diffusion • movement of molecules is not energy dependent • direction of movement is from high concentration to low concentration. With the concentration gradient • size of concentration gradient impacts rate of uptake

  25. Facilitated diffusion http://biocyc.org/ECOLI/new-image?type=ENZYME&object=GLPF-MONOMER. These proteins exist in plants, animals, fungi, bacteria, and protists.

  26. Facilitated diffusion… • Differs from passive diffusion • uses carrier molecules (permeases) • smaller concentration gradient is required for significant uptake of molecules • effectively transports glycerol, sugars, and amino acids • more prominent in eucaryotic cells than in procaryotic cells

  27. rate of facilitated • diffusion increases • more rapidly and • at a lower • concentration • diffusion rate • reaches a plateau • when carrier • becomes • saturated carrier saturation effect Figure 5.1

  28. note conformational change of carrier Figure 5.2

  29. Active Transport • energy-dependent process • ATP or proton motive force used • moves molecules against the gradient • concentrates molecules inside cell • involves carrier proteins (permeases) • carrier saturation effect is observed

  30. Transporters • “Molecular Properties of Bacterial Multidrug Transporters” – Monique Putnam, Hendrik van Veen, and Wil Konings – PubMed Central. Full Text available . Microbiol Mol Biol Review. 2000 December; 64 (4): 672–693

  31. ABC transporters • ATP-binding cassette transporters • observed in bacteria, archaea, and eucaryotes Figure 5.3

  32. antiport symport Figure 5.4

  33. Group Translocation • molecules are modified as they are transported across the membrane • energy-dependent process Figure 5.5

  34. Fe uptake in pathogens • The ability of pathogens to obtain iron from transferrins, ferritin, hemoglobin, and other iron-containing proteins of their host is central to whether they live or die • Some invading bacteria respond by producing specific iron chelators - siderophores that remove the iron from the host sources. Other bacteria rely on direct contact with host iron proteins, either abstracting the iron at their surface or, as with heme, taking it up into the cytoplasm

  35. Iron and signalling • Iron is also used by pathogenic bacteria as a signal molecule for the regulation of virulence gene expression. This sensory system is based on the marked differences in free iron concentrations between the environment and intestinal lumen (high) and host tissues (low) • Listeria Pathogenesis and Molecular Virulence Determinants José A. Vázquez-Boland,1,2* Michael Kuhn,3 Patrick Berche,4 Trinad Chakraborty,5 Gustavo Domínguez-Bernal,1 Werner Goebel,3 Bruno González-Zorn,1 Jürgen Wehland,6 and Jürgen Kreft3

  36. Pathogens and Iron uptake • Burkholderia cepacia • Campylobacter jejuni • Pseudomonas aeruginosa • E. coli • Listeria monocytogenes

  37. Iron Uptake • ferric iron is very insoluble so uptake is difficult • microorganisms use siderophores to aid uptake • siderophore complexes with ferric ion • complex is then transported into cell Figure 5.6

  38. Listeriosis • One involves the direct transport of ferric citrate to the bacterial cell • Another system involves an extracellular ferric iron reductase, which uses siderophores • The third system may involve a bacterial cell surface-located transferrin-binding protein

  39. Iron bacteria in the environment • There are several non-disease producing bacteria which grow and multiply in water and use dissolved iron as part of their metabolism. They oxidize iron into its insoluble ferric state and deposit it in the slimy gelatinous material which surrounds their cells. • These filamentous bacteria grow in stringy clumps and are found in most iron-bearing surface waters. They have been known to proliferate in waters containing iron as low as 0.1 mg/l.

  40. Culture Media • preparations devised to support the growth (reproduction) of microorganisms • can be liquid or solid • solid media are usually solidified with agar • important to study of microorganisms

  41. Synthetic or Defined Media • all components and their concentrations are known

  42. Complex Media • contain some ingredients of unknown composition and/or concentration

  43. Some media components • peptones • protein hydrolysates prepared by partial digestion of various protein sources • extracts • aqueous extracts, usually of beef or yeast • agar • sulfated polysaccharide used to solidify liquid media

  44. Types of Media • general purpose media • support the growth of many microorganisms • e.g., tryptic soy agar • enriched media • general purpose media supplemented by blood or other special nutrients • e.g., blood agar

  45. Types of media… • Selective media • Favor the growth of some microorganisms and inhibit growth of others • MacConkey agar • selects for gram-negative bacteria • Inhibits the growth of gram-positive bacteria

  46. Beta Hemolysis

  47. Types of media… • Differential media • Distinguish between different groups of microorganisms based on their biological characteristics • Blood agar • hemolytic versus nonhemolytic bacteria • MacConkey agar • lactose fermenters versus nonfermenters

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