9/3/2015

ITP 321 Lecture Note 2

Mi
Microbial
bi l Metabolisms
M t b li

Ratih Dewanti-Hariyadi
Department of Food Science and Technology
Bogor Agricultural University

Microorganisms metabolize substrate or

nutrient to yield energy and survive

CATABOLISM : ANABOLISM :

making cell materials, i.e.

cleaving reduced carbon to
yield energy using series of
enzymes or pathway
amino acid, protein, fatty
acid, nucleic acid
energy

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Microbial Metabolism

Organic waste product

Nutrients

Anabolism Catabolism

Energy
Cells Energy
Components

Energy source

Microbial Metabolisms
and Fermentation Technology

Microbes cell mass + metabolite

intermediate compounds

Fermentation technology
Block  Grow  High yield of 
mutant faster metabolites

Products amino acid biomass, acid, toxin, antibiotics

extra cellular
compounds

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Nutrient Transport System

• Simple Diffusion or Passive Diffusion

Substance movement due to difference in solute concentration, 
examples : water, O2, N
examples : water, O , N2
• Facilitated Diffusion or Carrier-mediated transport
Facilitated by permease or protein carrier
Energy is not required
Solute move from high to low concentration, e.g. glycerol
• Active Transport
Similar to facilitated diffusion but energy is required
Enable cell to accumulate solute at higher [conc] in
Enable cell to accumulate solute at higher [conc] in cytoplasm 
cytoplasm
Generate gradient difference which can be used for secondary active 
transport
•Group Translocation
Active transport that energy ultilizes generated from series of coupling 
reactions within microbial cells
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Nutrient Transport into Cell

Membran Inside
Outside Sitoplasma
1. Passive Diffusion; H2O H2O
Facilitated Diffusion Gly Gly

Lac Lac
2. Proton Symport H+ H+

Mel Mel
3. Sodium Symport
Na+ Na+

4 Active
4. A ti Transport
T t Mal Mal

APP+Pi

ATP

5. Group translocation : Man Man-P

Fosfotransferase System HPr
E1 PEP
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Active Transport

Outside

Membrane

Uniport Symport Antiport Inside

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Group Translocation Membrane

PEP E1 HPr‐P EIII

Glu‐P Glu

EII

Pyruvate E1‐P HPr EIII‐P

Inside Outside

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Aerobic and Anaerobic Metabolisms in

Microorganisms

• In the presence of [O2]

CHO  CO2 + H2O + [cell]  + ATP 
Aerobically, high amount of energy is produced

• In the absence of [O2]

CHO  C2H5OH + [cell]  + ATP 
Anaerobically, lower amount of energy is produced

• Facultatively anaerobic microorganisms can do both

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Catabolism And Energy

ATP
• an important intermediate
• compound with high energy PO4
• shows cells’ status

O O O NH2
   N C
HO – P – O – P – O – P – O C N
   
C C CH
OH OH OH CH2 N N
O H

H
Adenosine Tri Phosphate (ATP)

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Energy Status of Microbial Cells

ATP 0 5 ADP
ATP + 0.5 ADP
ATP + ADP + AMP

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Energy Status of Microbial Cells

ATP1 10 +5 ADP
ATP + 0.5 ADP0.5
0
____________________
ATP + ADP + AMP
11 ++ 10 ++ 01

Growing Cells has similar amount of ATP, ADP and AMP

ATP = ADP = AMP
Energy status is low (0.5)
Cells at the end of Growth Phase  ATP is very high
Energy status is high (1.0)

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Carbohydrate Catabolism
• Microorganisms oxidize carbohydrates as
th i primary
their i source off energy
• Glucose - most common energy source
• Energy obtained from Glucose by:
– Respiration
– Fermentation

Carbohydrate Catabolisms
maltose maltotriose
Outside cell

Protein
i Protein
P t i
carrier carrier cytoplasmic membran

maltose maltotriose
glucosidase Inside cell

glucose ‐ Phosphate

Aerobic Anaerobic

final electron acceptor :
cytoplasm
O2 organic /inorganic 

Final product
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Carbohydrate Catabolisms : Glycolysis

Glucose
ATP
hexokinase cytoplasmic membrane
ADP
Glu-6-P
glucosephosphate isomerase
glucosephosphate isomerase
Fructose-6-P
ATP
phosphofructokinase
ADP
Net : 2 ATP/glu
Fructose 1,6 di P

DHAP aldolase Glyceraldehyde ‐ 3 P

NAD glyceraldehyde‐P‐
NADH d h d
dehydrogenase
1,3 Diphospho Glyceric acid
ADP
phosphoglycerate kinase
ATP
3-phospho glyceric acid
PEP dehydratase
Phospho Enol Pyruvate
ADP
ATP pyruvate kinase
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Carbohydrate Metabolism: Pentose

Phosphate/Hexose Monophosphate Pathway
Glucosa

Gl
Glucose‐6‐P
6P

6‐P‐gluconate

Ribulose 5‐P
Ribose‐5‐P Xylulose‐5‐P

Glyceraldehyde-3-P Sedoheptulose‐7‐P

Fructose‐6‐P Erythrose‐4‐P

Fructose‐6‐P Glyceraldehyde-3-P
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Carbohydrate Metabolism: Entner

Doudoroff Pathway
Glucose

Glucose -6-P
NADP

NADPH
6-Phosphogluconate
phosphogluconate dehydrogenase

2-keto-3-deoxy-6-phosphogluconate

Glyceraldehyde-3-P Pyruvate

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Carbohydrate Metabolism: Phosphoketolase

Pathway (heterofermentative)

Glucose ATP D-ribose L-arabinose D-xylose

ADP ATP

Glucose-6-P ADP
NADP+
NADPH+H+
Glucololactone-6-P D-ribose 5-P L-ribulose D-xylulose

ATP ATP

6-phoshosfogluconate ADP ADP

NADP+
NADPH+H+
Ribulose 5-P

Xylulose 5-P
ADP ATP
Glyceraldehyde 3-P Acetyl-P Acetate
Pyruvate EMP

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Carbohydrate Metabolism :
Kreb’s/TCA Cycle
Pyruvat
CO2
ATP Asetil CoA

Citrate (C6)
NADH produced: 
Oxaloacetic acid
Aconitate ‐ for anabolism (glucose, protein 
Malate synthesis)
‐ converted into energy via 
Fumarate Isocitrate
oxidative phosphorylation (ETS)

Succinate

Oxalosuccinate
Succinyl co A

Oxoglutarate
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Carbohydrate Metabolism : Glyoxylate

Bypass (gowth on C2, hydrocarbon, FA)
Acetyl CoA

OAA Citrate

Malate malate synthase
Aconitate

Glyoxylate
i it t
isocitrate 
iyase

Fumarate
Isocitrate

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Aerobic Microorganisms

Possess Electron Transfer System (ETS) to transform NADH to ATP

NADH red ox red ox red O2

NADH-Q Cytochrom
QH2 Cy+Cl Cy+Cl
reductase oxidase

NAD
ox red ox red ox 2H2O

ATP ATP ATP

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Anaerobic Microorganisms

1. Some perform respiration using NO3, SO4 or Fe3+ as the final electron
acceptor (non-food!!)
2
2. Do not own ETS to convert NADH INTO 3ATP
3. Oxidize NADH in order to maintain metabolism
AH2 NAD BH2

catabolism

A NADH2 B organic compound
4. Carry out “substrate-level-phosphorylation

acetyl‐phosphate + ADP acetate + ATP (all anaerobes) 

carbamoyl + ADP carbamate + ATP (Clostridia)

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Alcohol Fermentation by Yeasts

EMP/ED*
Glucose Pyruvate
y

NAD NADH

CO2
Ethanol Acetaldehyde
CH3CH2OH CH3COOH

• Most yeasts : EMP

Zymomonas : ED

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Lactic Acid Fermentation by

Homolactic Bacteria

EMP
Glucosa Pyruvate

NAD NADH

Lactic Acid

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Propionic Acid Fermentation by

Propionibacterium

Piruvat Propionil CoA Propionat

Metil- OAA suksinat Suksinil CoA

malonil CoA

NAD NADH

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Butanadiol Fermentation

Pyruvate acetolactate + CO2

acetyl methy carbinol (acetoin)

NADH

NAD
2,3 butanadiol

Glycerol Fermentation

Triose – Phosphate Glycerol

NADH NAD

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Fermentation End Products

Lipid Catabolism

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Catabolism: Protein, Carbohydrate and Lipid

Q and A
1.  The main reaction in tape fermentation :
A. Mold cleaves carbohydrate into simple sugar
y p g
B. Yeast converts simple sugar into alcohol
C. Lactic acid bacteria converts alcohol into acetic acid
D. A,B correct
E.  A,B,C correct
2. The above yeast fermentation :
A. Is carried out aerobically to yield more ATP
B.  Is carried out anaerobically that utilize ETS
C.  Produces ethanol to reduce NADH
D. A,B correct
E. A,B,C correct

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