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Microbial MetabolismMicrobial Metabolism
Sofronio Agustin
Professor
Sofronio Agustin
Professor
LECTURES IN MICROBIOLOGYLECTURES IN
MICROBIOLOGY
LESSON 6LESSON 6
2
Lesson 6 TopicsLesson 6 Topics
Metabolism Energy Pathways Biosynthesis
3
MetabolismMetabolism
Catabolism
Anabolism
Enzymes
4
CatabolismCatabolism
Breakdown of complex organic molecules in order to extract energy and form simpler end products.
Enzymes are involved.
5
Metabolism ModelMetabolism Model
6
EnzymesEnzymes
Function Structure Enzyme-substrate interaction Cofactors Action Regulation
7
Enzyme StructureEnzyme Structure
Simple enzyme - primarily protein Conjugated enzyme
- protein and nonproteinThree-dimensional features:
Specificity -”lock-and-key”Active site or catalytic site
8
Conjugated EnzymesConjugated Enzymes
Conjugated enzymes contain a metallic cofactor, coenzyme, or both in order for it to function as a catalyst.
9
Active SiteActive Site
Specific active sites are folded regions of the protein molecule and contain specific amino acids in its microenvironment.
10
Enzyme-Substrate InteractionEnzyme-Substrate Interaction
Substrates specifically bind to the active sites on the enzyme:
-“lock-and-key” style
-Induced fit
Once the reaction is complete, the product is released and the enzyme reused.
11
Lock-and-Key ModelLock-and-Key Model
Specificity of enzyme-substrate reactions and induced fit.
12
Coenzymes Coenzymes
Function as transient carriers
Alter a substrate by removing a
chemical group from it and adding it
to another. Ex. NAD, FAD and CoA
13
Coenzyme ActivityCoenzyme Activity
Carrier function of
coenzymes
A coenzyme
transfers chemical
groups from one
substrate to another.
14
Enzyme ActionEnzyme Action
ExoenzymesEndoenzymesConstitutive
Induction or repressionTypes of reactions
15
Enzyme LocationEnzyme Location
Exoenzymes are inactive while inside the cell, but upon release from the cell they become active.
Endoenzymes remain in the cell and are always active.
16
Constitutive and Regulated EnzymesConstitutive and Regulated Enzymes
Constitutive enzymes are present in constant amounts.
Regulated enzymes are either induced or repressed.
17
Types of ReactionsTypes of Reactions
Condensation
Hydrolysis
Transfer reactions
18
Synthesis and Hydrolysis Synthesis and Hydrolysis
Condensation reactions are associated with anabolic reactions, and hydrolysis reactions are associated with catabolic reactions.
19
Transfer Reactions Transfer Reactions
Transfer of electrons from one substrate to
another.
Ex: Oxidoreductase - oxidation-reduction
reactions.
Transfer of functional groups from one molecule
to another.
Ex: Aminotransferases - transfer of amino group.
20
Sample Enzymes Sample Enzymes
Examples of enzymes, their substrates, and their reactions.
21
RegulationRegulation
Metabolic pathways
Direct control
Genetic control
22
Patterns of Metabolism Patterns of Metabolism
Metabolic pathways
follow stepwise
patterns.
These are regulated
by enzymes that
catalyze these
reactions.
23
Enzyme Control Mechanisms Enzyme Control Mechanisms
Competitive inhibition and noncompetitive inhibition are forms of direct control (regulation) of the enzyme action.
24
Genetic ControlGenetic Control
Repression - end products stop the expression of genes that encode for proteins (enzymes) which are responsible for metabolic reactions.
Induction - substrate initiates and enhances the expression of genes for proteins (enzymes) that drive metabolic reactions.
25
RepressionRepression
Repression as a type of genetic control of enzyme synthesis
26
Enzyme CharacteristicsEnzyme Characteristics
27
BioenergeticsBioenergetics
Cell energetics- Exergonic reactions
- Endergonic reactions
Redox reaction Electron carriers Adenosine Triphosphate (ATP)
28
Energy Machinery of the Cell Energy Machinery of the Cell
The general scheme associated with metabolism of organic molecules, the redox reaction, and the capture of energy in the form of ATP.
29
Redox ReactionRedox Reaction
Oxidation - removal or loss of electrons
Reduction - addition or gain of electronsThese are coupled reactions
Biological redox reactions involve transfer of
electrons and protons (hydrogens) =
dehydrogenationDehydrogenases - catalyze these reactions
30
Electron CarriersElectron Carriers
Electron carriers - transfer electrons (and
protons) from donor to acceptor molecules.
Coenzymes:
Ex: Nicotinamide adenine dinucleotide (NAD)
Respiratory chain (ETC) carriers:
Ex: Cytochromes (protein+porphyrin)
31
Adenosine TriphosphateAdenosine Triphosphate
Temporary energy repository (“cellular battery”)
Breaking of pyrophosphates bonds will release
free energy for cellular work.
Three part molecule:Nitrogen base - AdeninePentose sugar - Ribose)Chain of three phosphate groups
32
Energy CaptureEnergy Capture
The phosphate groups
capture the energy
derived from metabolism
as pyrophosphate bonds
within the ATP molecule.
ATP and its partner
compounds ADP and
AMP.
33
PhosphorylationPhosphorylation
ATP can be used
to phosphorylate
an organic
molecule such as
glucose during
catabolism.
Phosphorylation -
catalyzed by
phosphorylases(e.g. hexokinase)
34
Substrate-level PhosphorylationSubstrate-level Phosphorylation
ATP can be synthesized by substrate-level
phosphorylation.
A phosphate group from an intermediate is transferred to ADP to regenerate ATP.
35
Catabolic PathwaysCatabolic Pathways
Embden-Meyerhoff-Parnas (EMP) Pathway or Glycolysis
Kreb’s or Tricarboxylic Acid (TCA) Cycle
Electron Transport or Respiratory Chain
Alternate pathways
Fermentation
36
Glucose MetabolismGlucose Metabolism
Overview of the
location, flow, end-
products of cellular
(aerobic)
respiration.Glucose is catabolized to harness energy.
37
Cellular Respiration Cellular Respiration
Glycolysis
Kreb’s Cycle
Electron Transport Chain
38
Glycolysis Glycolysis
Glucose (6-carbon sugar) splits into two pyruvates (3-carbon molecules).
Glucose is oxidized and coenzyme NAD is reduced to NADH.
Energy investment phase:- Phosphorylation of intermediates using 2 ATP molecules
Energy yielding phase:- Substrate-level-phosphorylation of ADP to produce 4 ATPs.
39
Glycolytic StepsGlycolytic Steps
40
Kreb’s CycleKreb’s Cycle
Each pyruvic acid is processed to enter the Kreb’s Cycle as Acetyl CoA.
CO2 is generated -decarboxylation
reactions. Coenzymes NAD and FAD are reduced to
NADH and FADH2
Net yield of two ATPs per molecule of
glucose.
41
Steps in Kreb’s CycleSteps in Kreb’s Cycle
42
Electron Transport ChainElectron Transport Chain
•NADH and FADH2 from glycolysis and Kreb’s Cycle donate electrons to the electron carriers (ETC).•Membrane bound carriers transfer electrons by redox reactions.•Oxygen (final electron acceptor) completes the terminal step.
43
Electron Transport ChainElectron Transport Chain
The Electron Transport Chain and Chemiosmosis driven by the Proton Motive Force
44
Location of ETCLocation of ETC
Eukaryotes - Inner Mitochondrial
Membrane
Prokaryotes- Cytoplasmic Membrane
45
ATP YieldATP Yield
Glycolysis - 2Kreb’s Cycle - 2ETC- 34
Total Yield: 38
NADH yield - 2 in Glycolysis
8 in Kreb’s CycleFADH2 yield- 2 in Kreb’s Cycle
46
Anaerobic RespirationAnaerobic Respiration
Similar to aerobic respiration,
except nitrate or nitrite is the
final electron acceptor
47
FermentationFermentation
Glycolysis only
NADH from glycolysis is used to reduce the glucose
Organic compounds as the final electron acceptors (not O2)
Low ATP yields per glucose molecule compared to cellular respiration
48
FermentationFermentation
Chemistry of
fermentation:
Production of
ethyl alcohol or
lactic acid and
release of CO2
49
Types of FermentersTypes of Fermenters
Facultative anaerobes
Fermentation in the absence of oxygenRespiration in the presence of oxygen
Ex. Escherichia coli
Strict fermenters No respirationEx. yeast
50
Fermentation ProductsFermentation Products
Alcoholic fermentation
Acidic fermentation
Mixed acid fermentation
51
Mixed Acid FermentationMixed Acid Fermentation
Mixed acid fermentation and related products synthesized from pyruvate
52
BiosynthesisBiosynthesis
Amphibolic
Gluconeogenesis
Beta oxidation
Amination
Transamination
Deamination
Macromolecules
53
Amphibolic SynthesisAmphibolic Synthesis
Integration of the catabolic and anabolic
pathways (Coupled Reactions)
Intermediates serve multiple purposes
54
Amphibolic SynthesisAmphibolic Synthesis
Intermediates serve as precursors to synthesize amino acids, carbohydrates and lipids.
55
GluconeogenesisGluconeogenesis
Pyruvate (intermediate) is converted back to
glucose
Occurs when the glucose supply is low
56
Beta OxidationBeta Oxidation
Metabolism of fats into acetyl, which can
then enter the Kreb’s cycle as acetyl CoA.
57
Amino Acid SynthesisAmino Acid Synthesis
58
MacromoleculesMacromolecules
Cellular building blocks:MonosaccharidesAmino acidsFatty acidsNitrogen basesVitamins