MICROBIAL NUTRITION AND TRANSPORT

MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023

○ C, H, O, N ,S, P, K, Ca, Mg, Fe (exist as

cations)
GROWTH REQUIREMENTS ○ Sources:
■ Organic Compounds
■ Inorganic Salts
MICROBIAL GROWTH
● Increase in a population of microorganisms
Phospholipid Bilayer
● Due to the division of individual bacterial cells
Hydrophobicity of the
● Result of a microbial growth is a colony (arising
membrane is attributed to a
from one parent cell)
long chain of fatty acids made
○ Comprised/aggregate of microbial cells
out of carbon and hydrogen.
that arose from one parent cell
The hydrophilic ends are made
out of glycerol, phosphate, and
BIOCHEMICAL COMPONENTS OF CELLS
ethanolamine – we see carbon,
● Water: 80% wet weight
oxygen, phosphorus and even
● Dry weight
nitrogen which is needed for the
○ Protein: 40-70%
synthesis of the phospholipid
○ Nucleic acid: 13-34%
bilayer.
○ Lipid: 10-15%
○ Also monomers, intermediates, and
inorganic ions
Amino Acids Nucleic Acids

Glycogen

MACRONUTRIENTS
NITROGEN, PHOSPHOROUS, SULFUR
● All organic matter has C, H, & O but can also
MACRONUTRIENTS contain nitrogen, phosphorous, and sulfur
● Microbes need high amount of these nutrients Nitrogen
● Cells make protein, nucleic acids, and lipids ● Needed for the synthesis of amino acids,
● Macronutrients nucleotides, some sugars and lipids, and as
○ Play significant role in metabolism enzyme cofactors

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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023

●Many bacteria can use nitrogen from AA and

organic molecules MICRONUTRIENTS
● Some cells can utilize ammonia directly and
some reduce nitrite to ammonia.
MICRONUTRIENTS
● Some microorganisms can fix nitrogen
● Elements needed in trace quantities
(assimilate atmospheric nitrogen & reduce to
○ Co, Cu, Mn, Zn, V
NH4)
○ Enzymes
PHOSPHOROUS
○ Tap Water
● Required in nucleotides (also in ATP),
● Growth factors
phospholipids, cofactors, and some proteins
○ Organic compounds
● ALL bacteria can use inorganic phosphate
○ Vitamins
● Majority can incorporate or utilize phosphorus
directly
● Low amounts of phosphorous limit growth TRACE ELEMENTS
SULFUR ● Fe, Mn, Zn, Co, Mo, Ni, Cu
● Required for the synthesis of amino acids ● Required in trace amounts for some enzymatic
cysteine and methionine, some sugars, biotin, functions or protein stabilization
and thiamine ● In media culture, these are present in water or
● Supplied as sulfate or via organic sulfur media components or even by contaminants
compounds ● Ubiquitous in nature and are rarely growth
limiting
CO-FACTORS MACRONUTRIENTS
● K & Mg, Ca & Na for some microorganisms
● Cofactors for enzymes, stabilize ribosomes,
components of cytochrome, and
electron-carrying proteins

GROWTH FACTORS
● Compounds that are not synthesized by the
microorganism but important for growth and
metabolism
● Needs to be supplied in culture medium or
● Cofactors are important since many enzymes readily available in the environment
play significant roles in oxidation-redox ○ Amino Acids
processes (electron shuttling, etc.) ○ Purine + Pyrimidines
● Some enzymes like magnesium and potassium ○ Vitamins: enzyme cofactors
play significant roles in the proper function and
structure of the prokaryotic ribosomes
● Other enzymes act for stabilization of membrane
and DNA

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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023

Phototrophs undergo phototrophy because of their

energy source which is light. If the energy source is
METABOLIC DIVERSITY chemicals, the source is chemotrophs – can get from
organic chemicals (chemoorganotrophs) or inorganic
chemicals (chemolithotrophs)
METABOLIC DIVERSITY
Microbes can utilize carbon sources such as preformed
● Microbes are the most diverse when it comes to
organic molecules (heterotrophs) or CO2 (autotrophs).
metabolism

Plants can only produce ATP through photosynthesis

coupled with the calvin cycle and ETC. Animals
synthesize ATP through aerobic respiration using
glucose as the substrate via glycolysis and krebs
cycle and ETC. Microbes respire via different energy
sources: organic compounds such as glucose, light from
plants, inorganic compounds such as hydrogen sulfide.
When it comes to electron donors, not only water but
inorganic compounds. When it comes to terminal
electron acceptors, they have not only oxygen but nitrate
and also sulfate, and there are microbes that are EXAMPLES
fermentative. ● Photoautotrophs (Energy - Light, Carbon - CO2)
NUTRITIONAL MODE ○ Cyanobacteria
○ Purple sulfur bacteria
■ Splits H2S for electrons is
synthesis, producing sulfur.
● Chemolithotrophs (Energy - inorganic chemical
Carbon - CO2)
○ Nitrobacter winogradsky

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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023

NUTRIENT TRANSPORT: WHO GETS IN AND HOW?

NUTRIENT TRANSPORT

Left: Gram positive, Right: Gram Negative UPTAKE OF NUTRIENTS

● Positive: Transport of substances passes
through cell wall and across the cytoplasmic
membrane
● Negative: Substances pass through the outer
membrane going to the cytoplasm of the cell.
They pass across the amphipathic cytoplasmic
membrane. Sometimes, the nutrients pass
through the outer membrane and retain in the
periplasmic space to be degraded further by
degrading/hydrolytic enzymes.After degradation,
these are transported to the cytoplasm.

TRANSPORT OF NUTRIENTS IN THE CELL

● Nutrients are obtained from the environment and
must be specific
Group translocation and the ABC transport system
○ The cytoplasmic membrane is highly
are only present in bacteria.
selective except for gases because
these are really small via passive
diffusion. PASSIVE DIFFUSION
○ Many of the nutrients, in some form, are
usually against the concentration
gradient hence energy is supplied.
● Proteins embedded in the membrane
○ That’s why it’s asymmetrical
○ These proteins play a significant part of
the transport of nutrients.
● Transport against a concentration gradient
○ In the environment, nutrients are highly
diluted. Hence transport of nutrients ● Movement from high to low concentration
occur against the concentration ● Rate of diffusion is dependent on the size of
gradient. Energy is supplied. concentration gradient
○ Active transport (from a dilute solution) ○ Greater size/difference, the faster rate of
diffusion

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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023

● Oxygen, CO2, and H2O are transported into the SIMPLE TRANSPORT
cell by means of passive diffusion.
● Example: Osmosis
○ Referring to movement of water
molecules
● If we have the phospholipid bilayer, the blue
structures out the cell will move the compounds
into the cytoplasm.
FACILIATED DIFFUSION

With simple transport, it can be a symporter or antiporter.

A symporter refers to the protein embedded in the
cytoplasmic membrane that carries a substance. In
many cases, this is also a type of permease enzyme, it’s
just that there needs to be energy to transport the
substance. The lac-permease as a carrier protein
facilitates the entry of lactose. In the cytoplasmic
● Transport is facilitated through means of a
membrane, uses are permeability and energy
carrier protein/permease.
conservation. Proton motive force is in the cytoplasmic
● Carrier proteins can be specific or nonspecific,
membrane. This is a symporter because transport is
transporting a specific or many kinds of
unidirectional. For every proton that is translocated in the
substances
symporter, a molecule of lactose is delivered into the
● Transport proteins increase uptake rate which
cell. Usually for symporter and antiporter, there are 12
plateaus as the carrier protein saturates
helices. There is also a proton gradient with highly
● Size of concentration gradient impacts on the
saturated protons and negative ions. It acts like a battery
uptake rate of substances
to provide energy for use (chemiosmosis).
○ In passive diffusion, the rate is linear
In other cases, there is an antiporter. For every proton
compared to facilitated diffusion
entering the cell, sodium ions are discharged. This is
called the sodium-proton antiporter.There are different
THREE CLASSES OF TRANSPORT SYSTEMS directions – one enters, one exits the cell.
● Simple Transport
○ Driven by the energy in proton motive
force
● Group Translocation
○ Chemical modification of the transported
substance driven by a high energy
molecule phosphoenolpyruvate
● ABC Transporter:
○ Periplasmic binding proteins are
involved and energy comes from ATP

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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023
ABC SYSTEM (ATP BINDING CASSETTE)
● Energy dependent (ATP or proton motive force)
● Movement across a concentration gradient as
some nutrients are diluted in the environment.
● Carrier proteins – “permeases”
The ABC refers to the three protons needed in this type
of transport. A is for the periplasmic binding protein,
B for membrane-spanning transporter, and C for ATP
hydrolyzing protein. Here, the energy comes from ATP
or in some species, the proton motive force. How does
this happen?
In the periplasmic space, we have the periplasmic
protein wherein the solute is transported across the
peptidoglycan through simple transport, and then
another transport system will be used to deliver the
compound to the cytoplasm. This is only present in
GRAM NEGATIVE BACTERIA. For gram positive, there
might be a similar mechanism but is different from ABC.
The substance successfully entered the periplasmic
space, and the periplasmic binding protein carries the
transported substances and is delivered to the
cytoplasm. This substance needs to pass through the
cytoplasmic membrane. In the membrane, there is
another protein (Membrane-spanning protein) that will
facilitate entry of this substance. Then to release the
substance into the cytoplasm, another protein called
ATP-hydrolyzing protein that breaks down an ATP
molecule into ADP and inorganic phosphate so that the
transported substance is now released to the cytoplasm.
GROUP TRANSLOCATION/PHOSPHOTRANSFERASE SYSTEM
● Chemically modifies molecules as they are
being transferred into the cell.
● Transports sugars while phosphorylating them
using PEP (phosphoenolpyruvate)
● Modification of glucose sugar maintains
concentration gradient
● ONLY IN BACTERIA AND ARCHAEA
Here, a phosphate group is transferred from one enzyme
to another enzyme. We mentioned earlier that the
compound is chemically modified when it enters the cell.
Glucose needs to pass through enzyme IIC. Once it
enters the membrane, it is phosphorylated (phosphate
group is added to 6 carbon glucose). The phosphate
comes from 1 molecule of phosphoenolpyruvate. There
is a release of pyruvate molecules. This phosphate
group is translocated from one enzyme to another
enzyme called HPr, transferring to enzymes IIA and IIb for
phosphate to be added to glucose. Some components of
the phosphotransferase system are non-specific. This
means it's not only for glucose but other sugars. IIa and
IIb are specific. Many sugars are transported via group
translocation system such is the case for glucose and
mannitol.
IRON UPTAKE - SIDEROPHORES
● Ferric iron is insoluble
● Iron-chelating molecules
● Iron-siderophore complex reaches the cell surface
and binds to a siderophore receptor protein –
released to enter the cell directly – or enters via
ABC transporters
● Many pathogenic microbes have siderophores.
Recalling, when we talk about macro and micronutrients,
Iron is an important element for metabolism. These
pathogenic microorganisms get iron from red blood cell
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MICROBIAL NUTRITION AND TRANSPORT
MCROBIO (Lecture)
BIOMED | PROF. ESPIRITU | SEM 1 2023
as there is a heme ring made out of iron by means of
producing siderophores. They are secreted in
Pseudomonas as pyoverdine. Since these are
iron-chelating, these are complex with irons. These
siderophores bind to iron and goes back to the cell and
binds to a siderophore receptor protein present in the
cell.
MECHANISMS ON HOW CELL WOULD SECRET PROTEINS
● Microbes are able to secrete extracellular
proteins into the environment.
● Breakdown large compounds on the outside
● Secrete toxins during pathogenesis (e.g.
haemolysis)
● Some examples would be agarase, caraginase,
etc.
PROTEIN SECRETION
THE SEC AND TAT SYSTEMS
● Gram Positive: Protein must cross the plasma
membrane through a porous cell wall
● Gram Negative: Cross the plasma membrane,
relatively porous cell wall, and across the outer
membrane.
● Sec and Tat Pathways:
○ Secretion through cytoplasm membrane
○ Most proteins are secreted via
Sec-dependent secretion pathway
SEC-DEPENDENT PATHWAY
● Proteins to be transported across the membrane
exist as “pre-secretory” proteins
● Pre-protein has a signal peptide (N-Terminus)
recognized by SecA or signal recognition
particles
○ Normally, secA-exported proteins
secreted to periplasm while
SRP-exported are secreted to the
cytoplasmic membrane
○ ATP driven
○ When the pre-protein emerges outside
the membrane, a signal protein removes
the signal peptide (N-Terminus) and
protein folds into its normal
conformation.
DOC ESPIRITU’S EXPLANATION
As protein is being synthesized, the first few AA
synthesized are called the signal sequence. Signal
sequence functions as where Sec-A protein binds to.
When Sec-A protein binds to the signal sequence, the
fate of the protein is to be secreted to the periplasm. If
the protein is attached to signal recognition particle, the
fate of the protein is that it will be inserted into the
membrane (probably taking part in membrane secretion
system). We notice the protein is not fully folded. In TAT
pathway, the protein is folded before its secreted. If
protein does not contain signal sequence, it will not be
secreted.
PROTEIN SECRETION SYSTEMS OF GRAM NEGATIVE
Type I-VI Transport to the outside
Type I, III, IV & VI Form pores through both membranes
It forms a pore in the host membrane (protein is
directly injected – type III) or it can inject DNA
(type IV), or it can retract the needle forming a
pore in the host membrane (type VI). It helps the
microbe establish infection to a host cell by means
of secretion systems.
Type II and V Sec-dependent or tat-dependent
pathway used to secrete to periplasm
then protein secrete outside.
Before proteins are delivered into these systems,
these are delivered via the sec or tat system.