Microbial

metabolism
Bio 3213
Faculty of Natural Science
University of Guyana
  Introduction

 The Respiratory Chain

 Energy production by Anaerobic processes

Outline  Glycolysis
 The Pentose Phosphate Pathway
 ED pathway
 Fermentation

 Energy production by Aerobic processes

 TCA cycle
 The glyoxylate cycle
  Metabolism: all the organized chemical activates performed
by cell comprises of energy production & energy utilization

 Energy is used for :

1. Construction of the physical part of the cell
2. Synthesis of enzymes, NA, polysaccharides, & other
Introduction chm
3. Required for repair of damage
4. Growth & multiplication
5. Motility
6. For accumulation of certain nutrients in high conc. in
the cell
7. Keeping certain other substances out of the cell
  Sequences of oxidation-reduction rxns that occurs in cells

 The rxns are mediated by a no. of electron carriers & electron-

carrier enzymes
The
Respiratory  As the e- flow through the chains, much of their energy is
CONSERVED in the form of ATP (Oxidative phosphorylation)
Chain
(Electron transport chain)
 The multi-components of the ETC are always associated with
membranes:
 Eukaryotes - mitochondrial or chloroplast membranes
 Prokaryotes - cytoplasmic membranes
  Consists of enzymes having prosthetic groups or
coenzymes, each is an O/R system
The
Respiratory  The oxidized form of each prosthetic group/
coenzyme has an absorption spectrum different from
Chain that of the reduced from so that the 2 states can be
(Electron transport chain)
distinguished by spectrophotometry
  Certain enzymes which remove e- & H + from
reduced substrates (Dehydrogenases) have NAD+ &
NADP+ as their coenzyme

 NAD+ can exist in reduced form (NADH + H+) to

The Respiratory form an O/R system :
Chain :
COENZYMES NAD+ + 2 H+ + 2 e - NADH + H+
NAD & NADP
 In the same way, NADP+ can exist in a reduced state

 Niacin forms part of the structure of NAD & NADP &

is a precursor in their biosynthesis
  Another class of dehydrogenases (Flavoproteins)
exists & contains either FAD or FMN as prosthetic
groups
The
Respiratory  Riboflavin is a basic past of their coenzyme structure
Chain :  Can exist in either an oxidized or reduced form :
COENZYMES Riboflavin + 2 H+ Riboflavin-H2
FAD & FMN
 The reduced form of the coenzyme are FADH2 &
FMNH2
  Ubiquinone , a Quinone & is present in all cells

 A fat soluble coenzyme

The Respiratory  Functions as an acceptor of reducing power from the

Chain : Flavin-linked dehydrogenases:
COENZYMES Q
Flavoprotein-H2 + Ubiquinone

Ubiquinone-H2 + Flavoprotein
  NAD+ & NADP+ , Flavoproteins & Ubiquinones carry
2H+ & 2 e-
The Respiratory
Chain :  The cytochromes transfer only electrons
COENZYMES Q
 The protons being associated with an - NH2
group or a – coo- group & eventually transferred
to o2
  The prosthetic group of a cytochrome is a derivative
of HEME & contains a single iron atom which is
responsible for the oxidative or reductive properties
of the enzyme
The Respiratory
Chain :  On the basis of difference in absorption spectra,
CYTOCHROMES cytochromes can be divided into 3 main categories:
 Cytochrome a
 Cytochrome b
 Cytochrome c
  Each of these groups has a different function in the
respiratory chain & can be further subdivided on the
basis of minor differences in absorption spectra e.g.
Cytochromes c & c1 or cytochrome a & a3

The Respiratory  Each cytochrome type can exist in either an oxidized

Chain : or reduced form , depending on the state of the iron
CYTOCHROMES atom contained in their structure:

Cyt b-Fe3+ + e- Cyt b-Fe2+

Ferric Ferrous
  Act sequentially to transport e- from coenzyme Q to
O2
The Respiratory
Chain :  Cyt a & a3 = Cytochrome oxidase, both contain Cu
CYTOCHROMES
 Only Cyt a react directly with O2
  Sufficient energy for ATP synthesis is liberated at 3
points along the chain

 The incremental release of energy in the ETC results

The Respiratory in a more efficient trapping of energy in ATP than
Chain : direct oxidation of the reduced substrate of O2
Sequence of
oxidation  The RC of bacteria is associated with the cytoplasmic
membrane (Eukaryotes~ mitochondrial membranes)

 Much of the e- transfer in membranes is accomplished

within highly integrated particles or complexes
  e- transfer is accompanied by a flow of protons (H+)
from NADH2 through coenzyme Q but not in steps
The Respiratory involving cytochromes
Chain :
Sequence of  3 ATP are formed per molecule of NADH2 reoxidized
but only 2 ATP per molecule of FADH2 reoxidized
oxidation
 > 15 chm substances in the chain
 The
Respiratory
Chain
(Electron transport chain) P
P
 The
Respiratory
Chain
(Electron transport chain)
 The
Respiratory
Chain
 Energy  Heterotrophic bacteria can use a variety of
organic compounds as energy sources
production  CHO, organic & fatty acids & AA
by
Anaerobic  For many mo’s, the preferred compounds are
processes CHO especially the 6-carbon sugar glucose
  Most common pathway of glucose metabolism

 Embden-Meyerhof pathway

Glycolysis  Occurs very widely

(Splitting of sugar)

 Found in Mo’s and animals & plants

  Does not require the presence of O2
 Can occurs in both aerobic & anaerobic cells
Under Anaerobic conditions;
Fermentation
Glucose fermentation products
Glycolysis
(Splitting of sugar Under Aerobic conditions;
Fermentation Respiration
Glucose Intermediate CO2 + H20
(Pyruvate) O2
GLYCOLYSIS ~ STEPS 1, 3 & 10 ARE IRREVERSIBLE
  Fructose-1,6-diphosphate formed from glucose is split
into two 3-Carbon units (DHAP & glyceraldehyde-3-
phosphate) & they are subsequently oxidized to
pyruvic acid

Glycolysis  Where glyceraldehyde-3-phosphate is oxidized, a pair

of electrons (2 hydrogen atoms) is removed
(Splitting of sugar
 In the absence of O2, this pair of electrons may be
used to reduce pyruvic acid to lactic acid /ethanol

 In the presence of O2, this pair of electrons may

enter the respiratory chain
  Many of the rxns are freely REVERSIBLE & can be
used for the synthesis of glucose or for its breakdown

 Only 3 rxns are NOT REVERSIBLE by common

Glycolysis enzymes, but the presence of other enzymes can
reverse them for glucose synthesis to occur
(Splitting of sugar
 Hexokinase ~ step 1 (Phosphoenolpyruvate
synthase)
 Phosphofructokinase ~ step 3 (Phosphatases)
 Pyruvate kinase ~ step 10 (Phosphatases)
  For each molecule of glucose metabolized, 2
molecules of ATP are used up & 4 molecules of
ATP are formed.

Glycolysis  For each molecule of glucose metabolized, there

is a net yield of 2 ATP molecules
(Splitting of sugar

C2H12O6 + 2NAD + 2ADP + 2 Pi

2CH3COCOOH + 2NADH2 + 2ATP

(Inorganic pyruvate)
  Catabolic pathway

The  Exist in both prokaryotic cells & eukaryotic cells

Pentose
 It involves some rxns of the glycolytic pathway
Phosphate  Viewed as “SHUNT” of glycolysis = Hexose
Pathway monophosphate shunt (HMP shunt)
 Phosphogluconate pathway
(PPP)
 Glucose can be oxidized with the liberation of
electron pairs, which may enter the ETC
  Not considered a major energy-yielding pathway in
most Mo’s

The  Provides reducing power in the form of NADPH +

H+ , which is required in many biosynthetic rxns of cells
Pentose
Phosphate  Provides pentose phosphates for use in nucleotide
Pathway synthesis

 Provide energy for cell as an ALTERNATE PATHWAY

for the oxidation of glucose ~ also a mechanism for
obtaining energy from 5 –C sugars
  Involves the initial phosphorylation of glucose to form
glucose-6-phosphate which is oxidized to 6-
phosphogluconic acid with the simultaneous production of
NADPH
The
Pentose  Decarboxylation of 6-phosphogluconic acid together with
a yield of NADPH, produces ribulose-6-phosphate
Phosphate
Pathway  Epimerization rxns yield xylulose-5-phosphate & ribose-5-
phosphate which are the starting point for a series of
transketolase rxns & transaldolase rxns

 Leading subsequently to the initial compound of the

pathway (6-phosphogluconic acid) completing the cycle
  The 2 intermediates of glycolysis (fructose-6-
phosphate & glyceraldehyde-3-phosphate) are
generated
The
Pentose  The cell can carry out the complete oxidation of
Phosphate glucose-6-phosphate to CO2
Pathway
 Specifically, 6 molecules of ribulose-5-phosphate
& CO2 , 5 molecules of ribulose-5-phosphate
 The
Pentose
Phosphate
Pathway
 6 glucose-6-phosphate + 12NADP+

The 5 glucose-6-phosphate + 6 CO2 + 12NADPH + 12H + + Pi

Pentose
Phosphate
Pathway Net equation:
Glucose-6-phosphate + 12NADP+

6 CO2 + 12NADPH + 12H+ + Pi

 The  It is more probable that HMP hunt FEEDS INTO
Pentose THE GLYCOLYTIC PATHWAY by means of
Phosphate fructose-6-phosphate & glyceraldehyde-3-
phosphate
Pathway
  Another pathway of glucose metabolism

Entner-  Found in both aerobic & anaerobic

prokaryotes (NOT IN EUKARYOTES)
Doudoroff
pathway  IT IS FAIRLY WIDESPREAD particularly among
GNB
  Glucose if phosphorylated to glucose-6-phosphate
which is then oxidized to 6-phosphogluconic acid

 A dehydration step follows to yield 2-keto-3-deoxy-6-

Entner- phosphogluconic acid (KDPG), which is cleaved to
Doudoroff PYRUVIC ACID & glyceraldehyde-3- phosphate
 Metabolized by via some ED pathway enzymes to
pathway produce a 2nd molecule of pyruvic acid

 In the aerobic pseudomonads, the catabolism is

complete via acetyl CoA & TCA cycle
 Entner-
Doudoroff
pathway
Fermentation  Anaerobes also produce energy by fermentation which use
organic compounds as electron donors & acceptors

 Facultative anaerobes & obligate anaerobic bacteria employ

many kinds of fermentations to produce energy e.g. lactic
fermentation

 Streptococcus lactis is responsible for normal souring of milk,

dissimilates glucose to lactic acid which accumulates in the
medium as sole fermentation product ~ glycolysis
Glucose 2pyruvic acid Lactic acid
  In other CHO fermentations, the initial stages of
glucose dissimilation frequently follow the same
scheme of glycolysis
Fermentation
 Differences CHO fermentation occur in the ways the
pyruvic acid is used ~ variety of product may result

 PYRUVIC ACID IS THE HUB OF CHO FERMENTATION

 Most heterotrophic bacteria produce several end
products from glucose dissimilation but no single
species produce all these end products

Fermentation
Fermentation
  Mo’s can be grouped according to their products of
fermentation e.g. lactic acid group, propionic group

 Not ALL Mo’s metabolize the same substrate in exactly

Fermentation the same manner e.g. S. lactis & E. coli both ferment
glucose but by quite different pathways of
fermentation

 Some anaerobes do not have a functional glycolytic

system
 They may have CHO fermentation pathways that
use the HMP shunt & ED pathway
Group with E.g. of some Genera Products
Lactic acid bacteria Lactic acid only (Homofermentative) or lactic acid plus acetic acid,
- Streptococcus formic acid & ethyl alcohol (heterofermentative)
- Lactobacillus

Propionic acid bacteria Propionic acid plus acetic acid & CO2
- Propionibacterium
- Veillonella

Coli- aerogenes –typhoid bacteria Formic acid, acetic acid, lactic acid, succinic acid, ethyl alcohol,
- Escherichia CO2, H+ , 2,3-butylene glycol (various combination & amount
Fermentation
- Enterobacter depending on genus & species)
- Salmonella

Acetone, butyl alcohol bacteria Butyric acid, butyl alcohol, acetone, isopropyl alcohol, acetic acid,
- Clostridium formic acid, ethyl alcohol, H+, & CO2 (various combination &
- Bacillus amount depending on genus & species)

Acetic acid bacteria Acetic acid, gluconic acid

- Acetobacter
 Glucose

Glucose Succinic acid Pyruvic acid Lactic acid

fermentation
by E.coli Acetyl CoA Formic acid

Ethyl alcohol + Acetic acid CO2 + H +

 Glucose

Glucose
Pyruvic acid
fermentation
by S. lactis
Lactic acid
  TCA (citrate, cisaconitate & isocitrate) / Krebs cycle
Energy
production by  A sequence of rxns that generate energy in the
Aerobic form of ATP & reduced coenzyme molecules
processes (NADH2 & FADH2)

 Supply energy & provides many intermediates

Tricarboxylic required for the synthesis of AA, glucose, heme,
acid cycle etc.
(Citric acid cycle)  E.G. Oxaloacetic acid &  α- ketoglutaric acid are
AA precursors
  Involves the combination of a 2 C acetyl CoA with a 4C
oxaloacetate to produce a 6C tricarboxylic acid ,
CITRATE

Tricarboxylic  In the rxns that follow, the 2 carbons are oxidized to

acid cycle CO2 & oxaloacetate is regenerated & recycled
(Citric acid
cycle)  Oxaloacetate ~ play a catalytic role in this cycle

 Catabolic & anabolic in nature~ AMPHIBOLIC

Tricarboxylic
acid cycle
(Citric acid
cycle)
  Overall rxn :

Tricarboxylic
Acetyl CoA + 3H20 + 3NAD+ + FAD + ADP + Pi
acid cycle
(Citric acid
cycle)

2CO2 + CoA + 3NADH2 + FADH2 + ATP

  Used by some Mo’s when acetate is the sole carbon
source or during oxidation of primary substrates
(higher FA) that are cleaved to acetyl CoA w/o the
The intermediate formation of pyruvic acid
Glyoxylate
Cycle  Does not occur in higher organisms b/c they are never
forced to feed on 2- C molecules alone

 The specific enzymes are : isocitrate lyase & malate

synthase
  Used by some Mo’s when acetate is the sole carbon source or
during oxidation of primary substrates (higher FA) that are
cleaved to acetyl CoA w/o the intermediate formation of pyruvic
acid
The  Does not occur in higher organisms b/c they are never forced to
Glyoxylate feed on 2- C molecules alone

Cycle  The specific enzymes are : isocitrate lyase & malate synthase ~ fit
together with other rxns of the TCA to provide bypass around
some of the TCA – cycle rxns

 Overall rxn :
2 Acetyl-CoA succinate + 2H + 2CoA
 The
Glyoxylate
Cycle
  Acetyl CoA enters the cycle at 2 places

 Condenses with oxaloacetate to give citrate, which is the entry

point for the TCA cycle & the further rxn leads to the formation
of isocitrate
The  Isocitrate lyase is a splitting enzyme that produces succinate &
Glyoxylate glyoxylate
Cycle  The 2nd acetyl CoA condenses with glyoxylate to give malate by
the action of malate synthase

 Enzymes which carry out replenishment rxns such as this =

ANAPLEROTIC enzymes
 Function is to maintain the pool of essential intermediates for
biosynthesis