Functional anatomy of Bacteria

MANISH BANSAL

Comparing prokaryotic and eukaryotic cells



Prokaryotes
DNA is not enclosed in a membrane Lack membrane-bound organelles Cell walls contain peptidoglycan Reproduce by binary fission

Eukaryotes
DNA is surrounded by nuclear membrane Have many membrane-bound organelles Cell walls, when present, are usually simple Usually divide by mitosis

Proks and euks are similar in chemical composition and reaction

Proks lack membrane bound organelles Only Proks have peptidoglycan

Euks have membrane bound organelles Euks have paired chromosomes Euks have histones

Bacterial cell

Bacterial cell

The prokaryote
  

Unicellular Multiply by binary fission Classified by

Morphology Chemical composition Nutritional requirements Biochemical activates Sources of energy Other tests

Size
 

0.2 to 2um in diameter 2-8um in length In biological systems there are always exceptions these are general sizes.

Shape


Coccus
Diplococci Streptococci Staphylococci

  

Bacillus Spiral Other pleomorphic shapes

Cocci

Bacilli

Vibrio, spirillum, spirochete.

Structures external to cell wall



Glycocalyx
Capsule Slime layer

   

Flagella Axial filaments Fimbriae Pili

Parts not seen



Glycocalyx
Capsule Slime layer Extracellular polysaccharide

  

 

Function Toxicity

Protect from phagocytosis Allow adherence Reduce water loss Collect nutrients

Flagella
 

Used in movement Can present taxis

Negative Positive

 

Flagellar H protein acts as an antigen Flagellin

Flagella Arrangement

Figure 4.7

Flagella: Structure Long filamentous appendages with filament, hook and basal body

Fimbriae/pili


Shorter and less complex than flagella Helps adhere to surfaces Used for sex and communication

Cell Walls
Why study bacterial cell walls? They are essential structures in bacteria. They are made of chemical components found nowhere else in nature. They may cause symptoms of disease in animals. They are the site of action of some of our most important antibiotics.

 

 

Cell wall
  

Major difference between eukaryotic and prok orgs. Surrounds plasma membrane provides protection Peptidoglycan
Polymer of
  

NAG NAM Short amino acid chain

  

Prevents osmotic damage Damage to cw is almost always lethal except Production inhibited by antibiotics

Profile of the bacterial cell envelope

Gram-positive cell wall is thick homogeneous monolayer Gram-negative cell wall is thin heterogeneous multilayer

Cell wall


Gram-positive cell wall

Made of peptidoglycan in multiple layers (complex) Contains techoic acids

Gram-negative cell wall

Made of one layer of peptidoglycan (simple) No techoic acid is present Outer membrane with lipopolysaccharides Lipid portion of lipopolysaccharide is endotoxin

Structure of cell wall in gram positive bacteria

Gram Positives have thick cell wall and Teichoic acids

Gram negative


Lipoprotein phospholipid outer membrane surrounding a thin peptidoglycan Makes gram neg resistant to
Phagocytosis Antibiotics Chemical reactions Enzymes (lysozyme) Has lipid A endotoxin O polysaccaride antigen.

Gram-Negative Outer Membrane

Figure 4.13c

Chemical nature of bacterial cell walls

Bacterial cell walls always contain murein, which is a type of peptidoglycan Chemical nature of murein accounts for the function of the cell wall Murein is only found in the cell walls of bacteria
E. coli peptidoglycan

Chemical nature of bacterial cell walls

Peptidoglycan is made up of  2 amino sugars N-acetyl-glucosamine = G N- acetylmuramic acid = M


4 amino acids L-alanine = L-ala D-glutamic acid = D-glu diaminopimelic acid = DAP D-alanine = D-ala

Chemical nature of bacterial cell walls

Penicillin prevents formation of this Interpeptide bond

Lysozyme breaks this glycoside bond between M and G

Gram-negative murein showing the sites of action of the antibiotic penicillin and the enzyme lysozyme

Chemical nature of bacterial cell walls

Gram-positive murein has a thicker glycan a backbone and there are interpeptide bridges that join amino acid side chains together.

Chemical nature of bacterial cell walls

Penicillin blocks the Insertion of the interpeptide bridge

Lysozyme breaks the glycoside bond between M and G

Gram-positive murein showing the sites of action of the antibiotic penicillin and the enzyme lysozyme

Other characteristics of bacterial cell walls

Gram-positive cell walls contain teichoic acids

Teichoic acids are thought to stabilize the Gram positive cell wall and may be used in adherence.

Other characteristics of bacterial cell walls

Outer membrane of Gram-negatives has two important properties 1. It protects the cells from permeability by many substances including penicillin and lysozyme. 2. It is the location of lipopolysaccharide (endotoxin) which is toxic for animals.

Table: Correlation of the Gram stain with properties of bacterial cell walls
Property Thickness of wall Number of layers Peptidoglycan (murein) content Teichoic acids in wall Protein/lipoprotein content Lipopolysaccharide content Sensitivity to penicillin Sensitivity to lysozyme Gram-positive thick (20-80 nm) 1 >50% present 0-3% 0 sensitive sensitive Gram-negative thin (10 nm) 2-3 10-20% absent >50% 13 resistant resistant

Primary function of the bacterial cell wall



To prevent rupture or osmotic lysis of the cell protoplast

Lysis of a pair of dividing E. coli cells

Cell walls can be removed for genetic transfer



Protoplast
Gram-positive cell without cell wall

Spheroplast
Gram-negative cell without cell wall

Nontypical cell walls



Mycoplasma (acid fast) do not have ppt containing cell wall. Archaea contain another chemical called pseudomurein

Structures internal to cell wall



Plasma membrane
Selectively permeable fluid mosaic model Movement of materials across plasma membrane


Passive processes
Diffusion Facilitated diffusion Osmosis

Active processes
Active transport  Group translocation

Plasma membrane


Defines the living and nonliving parts of the cell

Everything on the inside is living Everything on the outside is not living

 

Is selectively permeable Workspace for enzymes of metabolic reactions

Cell (cytoplasmic) membrane



Completely encloses the bacterial cell protoplast Composed of 60% protein and 40% phospholipid Arranged as a bilayer

Section of a cytoplasmic membrane

Plasma Membrane
   

Phospholipid bilayer Peripheral proteins Integral proteins Transmembrane proteins

Figure 4.14b

  

Fluid Mosaic Model Membrane is as viscous as olive oil. Proteins move to function Phospholipids rotate and move laterally

Figure 4.14b

Membrane structure and assembly



The membrane bilayer is formed by phospholipidm olecules made up of glycerol and fatty acids

Membrane structure and assembly

Phospholipids arrange themselves spontaneously in water: lipid tails inward; glycerol heads outward

Transport systems in bacteria

Structures internal to cell wall



Cytoplasm
Nuclear area
 

Chromosomes Plamid(s)

Ribosomes Inclusions and granules

  

Metachromatic phosphate Sulfur Magnetosomes

Endospores

Membrane activity
     

Diffusion Osmosis Passive diffusion Facilitated diffusion Active transport Know the relationships

Movement Across Membranes

Active transport of substances requires a transporter protein and ATP. Group translocation of substances requires a transporter protein and PEP.

PM Workspace
Nutrient breakdown Energy production Photosynthesis Afforded by mesosomes which are regular infoldings of the plasma membrane


Weaknesses: destroyed by actions of alcohols, quaternary ammonium (detergents) and polymyxins antibiotics. Damage to the membrane causes leakage of cell contents.

Functions of the cytoplasmic membrane



Osmotic or permeability barrier: the membrane is impermeable to molecules that are charged or greater than molecular weight of 100 Location of transport systems to import all the needed molecules that are charged or greater than molecular weight 100 Energy generation: location of the electron transport system (ETS) and the ATP synthsizing enzyme ATPase Specialized functions involving cell wall synthesis, cell division and DNA replication.

Cytoplasmic Constituents of Bacterial Cells

 

 

Cytoplasm Genetic material: chromosome and Plasmids (DNA) Ribosomes Inclusions

Cytoplasm's


The liquid component of the cell within the PM Mostly water, dissolved ions, DNA ribosomes and inclusions Concept of homeostasis

Nuclear area
 

Contains the bacterial chromosome Bacteria may also have plasmids with up to 25% of the genetic materials

The Bacterial Chromosome or Nucleoid

Bacterial DNA released from a gently lysed E. coli cell

DNA (deoxyribonucleic acid) is the genetic material of the cell. It can be replicated in a semiconservative fashion and passed on to progeny cells.

Ribosomes

Figure 4.19

Ribosome Structure and Composition

Ribosomes are made of two subunits, a large subunit and a small subunit. Each subunit is made up of RNA and various proteins.

Ribosome Function

Ribosomes function in protein synthesis. Amino acids are assembled into proteins according to the genetic code on the surfaces of ribosomes during the process of translation.

Inclusions


    

Typically reserve deposits of excess materials like inorganic phosphate Polysaccharide granules Lipids Sulfur Gas iron

Small molecules present in a growing bacterial cell Molecules Amino acids, their precursors and derivatives Approximate number of kinds 120

Nucleotides, their precursors and derivatives Fatty acids and their precursors Sugars, carbohydrates and their precursors or derivatives quinones, porphyrins, vitamins, coenzymes and prosthetic groups and their precursors Ions (PO4, NH3, SO4, etc.)

100 50 250

300 20

Some inclusions in Bacterial Cells

Inclusion Glycogen Poly-betahydroxybutyric acid (PHB) Poly-phosphates Sulfur globules Magnetosomes Gas vesicles Parasporal crystals Composition poly-glucose lipid polymers of PO4 elemental S magnetite (iron oxide) protein shells inflated with gases protein

Function
Reserve carbon and energy source Reserve carbon and energy source Reserve phosphate, possibly high-energy PO4 Reserve energy and or electrons Provide orientation in magnetic field Provide buoyancy in aquatic environments Produced by endosporeforming Bacilli - toxic to insects

Some inclusions in Bacterial Cells

Bacterial Inclusions. A. PHB granules; b. a parasporal BT crystal in the sporangium of Bacillus thuringiensis; c. carboxysomes in Anabaena viriabilis, showing their polyhedral shape; d. sulfur globules in the cytoplasm of Beggiatoa.

Endospores


Resting and waiting stage Resistant to drying and other harsh conditions

Formation of spores

Endospores are produced as intracellular structures within the cytoplasm of certain bacteria, most notably Bacillus and Clostridium species.

Endospore forming bacteria left to right: Clostridium botulinum, Bacillus brevis, Bacillus thuringiensis

Under favorable nutritional and environmental conditions, an endospore germinates into a vegetative cell.

Medically-important Endosporeforming Bacteria

    

Bacillus anthracis causes anthrax Bacillus cereus causes food poisoning Clostridium tetani causes tetanus Clostridium botulinum causes botulism Clostridium perfringens causes food poisoning and gas gangrene Clostridium difficile causes antibiotic-induced diarrhea and pseudomembranous colitis

Properties of Endospores


Endospore formation is NOT a mechanism of reproduction. Rather it is a mechanism for survival in deleterious environments. During the process of spore formation, one vegetative cell develops into one endospore. Resting (dormant) cells - cryptobiotic i.e., show no signs of life..primarily due to lack of water in the spore Several unique surface layers not found in vegetative cells: exosporium, spore coat, cortex, and core wall Highly resistant to heat (boiling), acids, bases, dyes ( dont stain) irradiation, disinfectants, antibiotics, etc.

Properties of Endospores

Parasporal crystal

Endospore

Spores and parasporal crystals produced by some bacteria are toxic to insects