4 bio265 metabolism instructor dr di bonaventura
TRANSCRIPT
An overview of Microbial metabolism
What is Metabolism?
Catabolism (breakdown) Provides energy and building blocks for anabolism and other cellular functions
Anabolism (biosynthesis) Uses energy and building blocks to build larger molecules, macromolecules, and cell structures
Metabolism is composed of Catabolism and Anabolism
The sum of controlled chemical reactions that occur within a cell (microbial cell)
The role of ATP as an intermediate between catabolism and anabolism
Metabolic pathway - sequence of chemical reactions that occur in a cell
A B C D E
A is the initial molecule/compound (substrate) E is the final molecule/compound (product) B, C, and D are intermediates
Each step in the pathway is mediated or facilitated by
a specific catalyst called enzyme (protein)
Catabolic and anabolic reactions are organized into series of reactions called pathways
Cells use redox reactions to extract energy from nutrient molecules such as glucose
Redox reaction (oxidation-reduction reaction)
Redox reactions involve the transfer of electrons from an electron donor to an electron acceptor
The energy of the reduced coenzyme NADH (or FADH2) is used to
make ATP in later reactions
Cells use redox reactions to extract energy from nutrient molecules such as glucose
Most biological oxidations involve the loss of hydrogen atoms (one electron plus one proton-H+) - dehydrogenation reactions
Enzymes are the catalysts of biological reactions
Specific for a given substrate/chemical reaction
The shape of the molecule provides a distinctive site called the active site or catalytic site of the enzyme
The substrate specifically fits into the enzyme’s active site
Examples of enzymes and their names
Enzymes can use cofactors and coenzymes
Cofactors include magnesium, manganese, iron, copper, zinc, calcium, cobalt
Coenzymes (NAD+, NADP+, and FAD derived from vitamins) act as electron carriers
Enzymes lower the activation energy needed to “trigger a chemical reaction”
The substrate interacts with the active site of the enzyme to form an enzyme-substrate complex
The substrate is transformed into product(s) The product(s) is/are released The enzyme is recovered unchanged
Factors that influence the activity of enzymes include Temperature pH Enzyme and substrate concentration Presence/absence of inhibitors
Temperature and pH denature proteins, therefore enzymes
Inactive
Cells control synthesis of enzymes (amount/time of synthesis) and their activity
Enzyme inhibitors
Competitive inhibition
Sulfa drugs are an example of competitive inhibitors
Sulfanilamide/PABA
Inhibition can be reversible or irreversible
Noncompetitive inhibition
Poisons such as fluoride are an example of noncompetitive inhibitors
Enzyme inhibitors
Inhibition can be reversible or irreversible
Feedback Inhibition
It is reversible
When the final product accumulates
It begins to bind to and inactivates the enzyme that catalyzes the first reaction of the pathway
The pathway is turned off
Cells regulate metabolic
pathways
Carbohydrate Catabolism
Most microorganisms oxidize carbohydrates as the primary source of cellular energy
Two general processes are used Cellular respiration Fermentation
Both cellular respiration and fermentation can share a common pathway called glycolysis or Embden-Meyerhof pathway
Aerobic cellular
respiration Glucose (C6H12O6) is oxidized to CO2 in presence of O2
~ 38 molecules of ATP are formed
Fermentation
Releases energy from oxidation of organic molecules, i.e.; Sugars Organic acids Amino acids Purines and pyrimidines
Does not use the Krebs cycle or the electron transport chain
Uses pyruvate as the final electron acceptor from NADH, regenerating NAD+ for glycolysis
Does not require oxygen
End-Products of Fermentation
Chemical analyses of the end-products help identify microbes, including pathogens in clinical specimens
Lactic acid and alcohol Fermentation
Lactic acid bacteria are Streptococcus and Lactobacillus
Alcohol fermentation carried out by Saccharomyces cerevisiae (yeast)
How do we use this knowledge to distinguish between bacteria in the lab?
Fermentation test tubes containing mannitol
Negative control
Samples
S. epidermidis (-) S. aureus (+) E. coli (+) plus gas Bubbles in the Durham tube indicate gas formation
Microorganisms catabolize complex sugars and lipids
Microbes produce extracellular enzymes Amylases to break down
starch
Lipases to break down fats into glycerol and fatty acids
Microorganisms catabolize proteins
Microbes produce proteases
Break down proteins into their component amino acids, transported across the plasma membrane
Amino acids are chemically modified by deamination reaction
How do we use this knowledge to distinguish between bacteria in the lab?
Biochemical tests are designed to detect the presence of enzymes The test tubes contain
glucose pH indicator amino acid
Negative control Sample
pH indicator turns to yellow if acid is produced from glucose pH indicator turns to purple if alkaline products are produced from
amino acid (decarboxylation reactions)
How do we use this knowledge to distinguish between bacteria in the lab?
Production of H2S
Salmonella (+) can be readily distinguishable from E. coli (-) by the production of hydrogen sulfide
H2S is detected when bacteria remove sulfur from certain amino acids
Other tests take advantage of different components of the electron transport chain (oxidase test – cytochrome oxidase)
Anabolic pathways and their link to catabolic pathways
ATP made during catabolic pathways is used for cellular functions such as Active transport across plasma membranes Flagellar motion
Most of the ATP is used to synthesize new cellular components Amino acids are needed to make proteins/enzymes Carbohydrates needed for polysaccharides/peptidoglycan Lipids are important components of cell membranes Purines and pyrimidines are the building blocks of DNA and
RNA
Anabolic pathways and their link to catabolic pathways
The biosynthesis of simple lipids
Glycolysis and Krebs cycle provide intermediates (precursor metabolites) for anabolic pathways
Metabolic pathways that function in both anabolism and catabolism are called amphibolic pathways
Amphibolic pathways