Ch. 4 Nutrition, Culture, and Metabolism of Microorganisms: Monday, July 27, 2015 9:33 PM

Ch.
4 Nutrition, Culture, and Metabolism of

Microorganisms
Monday, July 27, 2015
9:33 PM
Page 1
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Page 3
the sum total of all chemical reactions that occur in a cell
energy-releasing metabolic reactions
energy-required metabolic reactions
Page 4
supply of monomers (or precursors of ) required by cells for growth
nutrients required in large amounts

amount are not based on molecular weight
nutrients required in trace amounts
Figure in the book

macromolecular composition of a cell is
protein = 55% (cell is a bag of enzymes)
Page 5
- typical bacteria cell ~50% carbon (by dry weight)

- major elements in all classes of macromolecules
Essential elements as a percent of cell dry weight

C = 50%
Page 6
typical bacterial cell ~13% nitrogen (by dry weight)

key element in proteins, nucleic acids, and many more cell
constituents.
Page 7
- synthesis of nucleic acids and phospholipids
- sulfur-containing amino acids (cysteine and methionine)

- vitamins (e.g. thiamaine, biotin, and coenzyme A
- required by enzymes for activity
Page 8
- stabilizes ribosomes, membranes, and nucleic acids
- helps stabilize cell walls in microbes

- plays a key role in heat stability of endospores
- required by some microbes

(marine microbes in sea water)
which of the following not a macronutrients?

iron is a between depending on the oragnism and environmental
condition
Page 9
key component of cytochromes and FeS proteins involved

in electron transport
insoluble form
IRON
siderphores
Fe 3+ (Ferric): insoluble
under oxic condition
Fe 2+ (Ferrous): soluble
under anoxic condition
Cells produce siderphores (an iron-binding
agents) to obtain iron from insoluble mineral form
(Figure 4.2)
soluble
Table 4.1 - you can see Iron(Fe)b is a

macronutrient as well.
Page 10
-organic compounds required in small amount by certain

organisms (vitamins, amino acids)
: most commonly required growth factors
Page 11
precise chemical composition is known

even the concentration
composed of digests of chemically undefined substances

(e.g. yeast and meat extracts)
ex: TSA and TSB
Table 4.2
Major components of Yeast Extract
Page 12
- contain compounds that selectivity inhibit growth of

some microbes, but not others
ex: antibiotic resistance
- contain an indicator, usually a dye, that detects particular

chemical reactions occurring during growth.
Page 13
can gram positive ferment?
example for both selective (gram negative) and differential

media (fermentation)
MacConkey agar
contains:
E.coli
Salmonella
bile salt
G(+) = don't grow
lactose,
peptone
both bacteria are G(-)

pink color = E. coli
lactose to produce lactic acids
(pH <7) [pink] = fermentation
no color = Salmonella
peptone to produce ammonia (pH >7)

[no color change] = no fermentation
agar is differential medium to differentiate G(+) and G(-)

= false
Page 14
it is important to know the nutritional requirements and

supply them in proper form and proportions in a culture
medium.
Page 15
culture containing only a single kind of microbe
unwanted organisms in a culture.
agar
colonies
Page 16
distinguish from
each other!
absence of all
microorganisms as well
as viruses
Sterilization
Aseptic technique
autoclave at 121 C for

15 - 20 minutes
manipulation to prevent
contamination of sterile objects of
microbial cultures during handling
Page 17
hood vent = air flows from in to out
7/28/2015 12:28 PM
Page 18
pour plate
Erlenmeyer flask of culture
autoclave medium, let cool for
40-50 C
pour on a plate and incubate
Page 19
spread plate
pipet 100-200 L
inoculate onto agar plate
used hockey stick to spread
(glass)
streaking
Question: any gram positive bacteria doing fermentation?
yes! yogurt Streptococcus thermophilus and Lactobacillus
acidophilus or L. bulgaricus
Page 20
7/29/2015 10:50 AM
conceptual
Page 21
energy is released that is available to work.
even metabolism produces heat
Page 22
- release free energy (exergonic)
- require energy (endergonic)
Page 23
A + B --> C + D
G0' = Gf* [C+D] - Gf* [A+B]

G < 0 (-G) --> [A + B] higher energy, energy released = exergonic
G < 0 (+G) --> [C + D] higher energy, energy required = endergonic

practice at home on a clean sheet of paper!
Page 24
only reveal whether

energy is required or
released.
energy required to bring all molecules in a chemical
reaction in to the reactive state (Figure 4.6)
Page 25
substance that lowers the activity energy of reaction
i. increases reaction rate

ii. does not affect energetic or equilibrium of a reaction
does not affect G
red line = without catalyst

high activation energy
green line = with catalyst
low activation energy
Page 26
- biological catalysts
- typically proteins
some RNAs (e.g. ribozyme)
- highly specific
- generally larger than substrate
- typically rely on weak bonds (not covalent bonds)
examples: hydrogen bonds, van der Waals forces,
hydrophobic interactions
Active site: the region of enzyme that binds to substrate
Page 27
spontaneous rate without

enzyme
E + S ----> E - S --->
E+P
E: enzyme
S: substrate
P: product
- substrate binding
- position of substrate relative to catalytically active
amino acids in active site
Page 28
E-S
Page 29
- bind tightly to enzymes

- usually bind covalently and permanently
- (e.g., heme groups in cytochromes)
- loosely bound to enzymes

- most are derivatives of vitamins
(e.g. NAD+ / NADH)
NADH = electron donor
Page 30
Page 31
energy-rich compounds
(e.g. ATP)
the substance oxidized in a redox reaction
the substance reduced in redox reaction
Page 32
is H2
is O2
N2 (N N)
most stable form.
H2 + O2 ---> H2O
H = electron donor
O = likely to accept [very selfish, hungry for electrons]
H2 --> 2 H+ + 2 e-
electron donating half reaction
2 H+ + O2- --> H2O
formation of water
O2 + 2 e- ---> O2-
Page 33
electron accepting half reaction
tendency of a compound to donate

electrons
- measured under standard conditions expressed as volts (V)

1 ATM
25 C
all reactions and products are 1 M concentrations
pH 7
Page 34
represents the range of possible

reduction potentials
0.04
0.84
1.24
left/ right (electron acceptor/electron donor)

oxidized substance/ reduced substance
Page 35
understand the table!!!

the more drop in the
redox tower, the
greater the amount of
energy released.
based on the organisms

given, know how to
calculate which
produces more energy!
0.42
E0' (V)
0.02
(1) 0.44
(2) 0.84
(3) 1.24
0.42
no3/no2
/donor
0.82
Fe3+/Fe2+
acceptor/
Page 36
carriers
(attached to enzymes)
(diffusible)
examples: NAD+, NADP (Figure 4.10)

NAD+, NADH = "redox couple"
Page 37
+ = receive electrons
H = donate electrons
NAD+ NADH + H+
oxidized (receiving electron) ---> reduced (donating electron)
Page 38
(E-S)
(S)
reaction I
NAD+ =
acceptor
substrate =
donor
electron
acceptor
reaction 2
NADH =
donor
substrate
=
acceptor
the substrate is
reduced
Page 39
energy-rich compounds
Page 40
- examples in prokaryotes
glycogen
poly--hydroxybutyrate (PMB) and other polyhydroxyalkanoates
- examples in eukaryotes
starch
lipids (simple fats)
Page 41
Page 42
1 glucose --->
6 carbons
2 pyruvate
2 * 3 carbons
ex: human
differ in mechanism of ATP synthesis
substrate-level phosphorylation; ATP directly synthesized
from an energy-rich intermediate
oxidative phosphorylation: ATP produced from proton

motive force (PMF) formed by transport of electrons
involved in (ETC)
through fermentation, ATP can be oxidative phosphorylation = false
Page 43
Fermentation
Respiration
(PMF)
3 H+ ---> ATP
respiration is more efficient than fermentation!
Page 44
fermentation - there is no TEA (terminal electron acceptor)
C6H12O6 ---> pyruvate
Page 45
Page 46
1 glucose ---> 2 pryuvate

- glucose consumed
- two ATPs produced
by substrate-level phosphorylation
- fermentation products generated
- glycolysis is part of fermentation
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Ch. 4 Nutrition, Culture, and Metabolism of Microorganisms: Monday, July 27, 2015 9:33 PM

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Ch.

4 Nutrition, Culture, and Metabolism of

the sum total of all chemical reactions that occur in a cell

energy-releasing metabolic reactions

energy-required metabolic reactions

supply of monomers (or precursors of ) required by cells for growth

nutrients required in large amounts

Figure in the book

- typical bacteria cell ~50% carbon (by dry weight)

Essential elements as a percent of cell dry weight

typical bacterial cell ~13% nitrogen (by dry weight)

- synthesis of nucleic acids and phospholipids

- sulfur-containing amino acids (cysteine and methionine)

- required by enzymes for activity

- stabilizes ribosomes, membranes, and nucleic acids

- helps stabilize cell walls in microbes

- required by some microbes

which of the following not a macronutrients?

key component of cytochromes and FeS proteins involved

Table 4.1 - you can see Iron(Fe)b is a

-organic compounds required in small amount by certain

: most commonly required growth factors

precise chemical composition is known

composed of digests of chemically undefined substances

Major components of Yeast Extract

- contain compounds that selectivity inhibit growth of

- contain an indicator, usually a dye, that detects particular

can gram positive ferment?

example for both selective (gram negative) and differential

both bacteria are G(-)

peptone to produce ammonia (pH >7)

agar is differential medium to differentiate G(+) and G(-)

it is important to know the nutritional requirements and

culture containing only a single kind of microbe

unwanted organisms in a culture.

autoclave at 121 C for

hood vent = air flows from in to out

energy is released that is available to work.

even metabolism produces heat

- release free energy (exergonic)

- require energy (endergonic)

G0' = Gf* [C+D] - Gf* [A+B]

G < 0 (+G) --> [C + D] higher energy, energy required = endergonic

only reveal whether

substance that lowers the activity energy of reaction

i. increases reaction rate

red line = without catalyst

Active site: the region of enzyme that binds to substrate

spontaneous rate without

- bind tightly to enzymes

- loosely bound to enzymes

the substance oxidized in a redox reaction

the substance reduced in redox reaction

electron donating half reaction

2 H+ + O2- --> H2O

electron accepting half reaction

tendency of a compound to donate

- measured under standard conditions expressed as volts (V)

represents the range of possible

left/ right (electron acceptor/electron donor)

understand the table!!!

based on the organisms

examples: NAD+, NADP (Figure 4.10)

oxidized (receiving electron) ---> reduced (donating electron)

oxidative phosphorylation: ATP produced from proton

through fermentation, ATP can be oxidative phosphorylation = false