Bacterial Cell Structure and Function

The Three Domains of Life Microbiological focus Archaea The Extremophiles Bacteria- Cyanobacteria and eubacteria Eukarya Protozoans, fungi, and worms

Prokaryote and Eukaryote Cells

The Structure of the Prokaryote Cell Small size ( 0.5 to 2um) Large surface area to volume ratio A variety of shapes Outer cell wall- very thick made of specialized molecules Cell membranes may have a different constituency of molecules from eukaryote cells Ribosomes smaller No organelles, no nuclear membrane 1 ds circular loop of DNA

Shapes of the Bacteria

Bacillus Coccus Vibrio Spirillum Spirochete Pleiomorphism- Some bacterial shapes vary within a culture. This can occur when the nutrients are used and wastes have built up

Bacterial Shapes

Bacillus- E. coli

Fluorescent stain

Freeze fracture

Public enemy # 1- Anthrax

Streptococcus pyogenes- strep throat

Staphylococcus

Arrangement of bacteria
Groups of two- diplo Chains- strepto Large groups- like grape clusters- staphylo Division on 3 planes- sarcinae- 8 cells arranged in a cube Division on two planes produces tetrads

Cell Morphology

Staphylococcus

Diplococcus

Bacterial Cell Structure and Function

The Three Domains of Life Microbiological focus Archaea The Extremophiles Bacteria- Cyanobacteria and eubacteria Eukarya Protozoans, fungi, and worms

Prokaryote and Eukaryote Cells

The Structure of the Prokaryote Cell Small size ( 0.5 to 2um) Large surface area to volume ratio A variety of shapes Outer cell wall- very thick made of specialized molecules Cell membranes may have a different constituency of molecules from eukaryote cells Ribosomes smaller No organelles, no nuclear membrane 1 ds circular loop of DNA

Shapes of the Bacteria

Bacillus Coccus Vibrio Spirillum Spirochete Pleiomorphism- Some bacterial shapes vary within a culture. This can occur when the nutrients are used and wastes have built up

Bacterial Shapes

Bacillus- E. coli

Fluorescent stain

Freeze fracture

Public enemy # 1- Anthrax

Streptococcus pyogenes- strep throat

Staphylococcus

Arrangement of bacteria
Groups of two- diplo Chains- strepto Large groups- like grape clusters- staphylo Division on 3 planes- sarcinae- 8 cells arranged in a cube Division on two planes produces tetrads

Cell Morphology

Staphylococcus

Diplococcus

Streptobacilli

Spirillum

Spriochetes

 Peptidoglycan the single most important molecule in the cell walls of bacteria One immense- covalently linked molecule The molecule forms a chain- the constituents are sugar-amino molecules( glucosamines)

Peptidoglycans - Cell Wall

Gram Positive Bacteria- Cell walls The cell wall is made of Teichoic acid- glycerol+ phosphates+ribotol which an alcohol sugar. These polymers extend beyond the cell even beyond the capsule

Gram Negative Bacteria- Cell Walls The outer membrane found primarily in Gramnegative bacteria is a bilayer that forms the outermost layer of the cell wall and is attached to the peptidoglycan by lipoproteins molecules.

The Cell Membrane LPS- this is actually a part of the outer cell membrane and can b be used to help to identiry Gram- bacteria. It is also important to the cell wall and is usually not released until a bacterium is dead. It consists of polysaccharides and lipid A

The toxins gram negative bacteria release are from this portion of the cell membrane. Gram negative bacteria release endotoxin and the result is fever, the dilation of blood vessels, so the blood pressure drops and causes other related effects.

Gram-Positive Bacteria The cell wall has a thick layer of peptidoglycan 20 to 80um thick 60-90% of the cell wall is peptidoglycan Except for the strptococci most gram positive bacterial cell walls contain very little protein

Gram-negative Bacteria The wall of a Gram-negative bacterium is thinner but more complex than a Grampositive bacterium Only10 to 20 % of the cell wall is peptidoglycan- the remainder consists of various polysaccharides, protein, lipids. The cell wall contains an outer membranethe LPS

The Periplasmic Space

The periplasmic space is between the outer surface of the cell wallo and the cell membrane Enzymes and toxins remain in the periplasmic space in sufficient concentrations to help destroy substances that might harm the bacterium.

The Cell Membrane

The cell membrane consists of molecules called phospholipids. Phospholipids have two long tails consisting of hydrocarbon chains ( HYDROPHOBIC) Phospholipids have a phosphate head (HYDROPHILIC) The membrane is formed by a double layer of these molecules The membrane also contains PROTEINS

Internal Structure Ribosomes- consist of ribonucleic acid ( RNA) and protein. They are abundant in the cytoplasm as POLYRIBOSOMES

Nucleoid Region- Central The nucleoid region contains 1 ds ciruclar loop of DNA. The DNA may be attached to the cell membrane as well at some point There may be RNA associated with the DNA

Chromatophores- photosynthetic bacteria

Molecules of life

Chromatophores are derived from the cell membrane. They contain pigments used to capture light energy for the synthesis of sugars. Nitrifying bacteria also may have these internal membranes. They contain the enzymes necessary for the energy transformation process of photosynthesis

The Nucleoid Region

Mesosomes- Bacteria Bacterial cells have large infoldings in their membranes.

Mesosomes Mesosomes provide the surface area for all of the chemical reactions in the cell respiration metabolic processes
Freeze fracture

Inclusions

Small bodies in the cytoplasm- some are called granules

Glycogen pyrrophosphate granules- volutin Metachromatic granules Variable colors

 Vegetative cells of bacteria like Bacillus or Clostrium produce

resting stages These spores are designed for survival and not reproduction. These are formed within cells They are resistant to heat, drying,acids,bases,disinfectants, and radiation

Spore formation Spores form when nutrients are depleged form a culture Few spores are formed when nutrients are plentiful and environmental conditions are favorable.

Spore Structure

An endospore consists of a core, surrounded by a cortex, a spore coat and in some species a thin layer called the exosporium spore

How do spores survive ?

They contain dipicolinic acid and a large number of calium ions. These materials contribute to heat resistance The lose water content enables them to survive.

 Endospores have survived over 10,000 years Special methods may be used to skill them during sterilization When conditions are favorable they can still germinate

Flagella

Bacteria that are motile have appendages called flagella A bacteria can have one or many flagella monotrichous amphitrichous lophotrichous peritrichous

What is this type of bacteria ?

 The diameter of a prokaryotes flagellum is about one-tenth of that of a eukaryote It is made of flagellin The basal region has a hook like structure and a complex basal body The basal body consists of a central rod or shaft surrounded by a set of rings Gram negative bacteria have a pair of rings embedded in the cell membrane and another pair of rings associated with the peptidoglycan and lipopolysaccharide layer of the cell wall. Gram positive have one ring.

Turns like a mixer

Flagellar motion Flagella rotate like twirling L- shaped hooks such as a dough hook on a kitchen rotor- or a mixer

Twiddling ? HHMM!

When flagella bundle together they rotate counterclockwise and the bacteria run

When the flgella rotate clockwise the flagellar bundle comes apart and the bacteria twiddle- tumble randomly

Chemotaxis

Sometimes bacteria move toward or away from substances in their environment. This is called chemotaxis Concentrations of most molecules in the environment form a gradient. When a bacteria is running a long an increasing gradient if reduces the frequency of its twiddles

PILI

Attachment piliThese are structures on the surface of the bacteria that aid the bacterium in atttching to surfaces These acid the bacterium by allowing colonization of the mucus membranes of organisms. Some bacteria adhere to red blood cells by attachment pil and cause red blood cells to clum- this is hemagglutination

SEX PILI- Conjugation

Conjugation pili or sex pili are found only in certain groups of bacteria. This structure exists exclusively for the transfer of DNA between bacteria The DNA passes between bacteria tend to develop antibiotic resistance

Bacterial Slime Layer or Capsule

Structure of Bacteria

Size of Bacteria
Average bacteria 0.5 - 2.0 um in diam. Surface Area ~12 um^2 Volume is ~4 um Surface Area to Volume is 3:1 Typical Eukaryote Cell SA/Vol is 0.3:1 Food enters through SA, quickly reaches all parts of bacteria Eukaroytes need structures & organelles
RBC is 7.5 um in diam.

Shapes of Bacteria
Coccus
Chain = Streptoccus Cluster = Staphylococcus Chain = Streptobacillus

Bacillus

Coccobacillus Vibrio = curved Spirillum Spirochete Square Star

Bacterial Structures
Flagella Pili Capsule Plasma Membrane Cytoplasm Cell Wall Lipopolysaccharides Teichoic Acids Inclusions Spores

Flagella
Motility - movement Swarming occurs with some bacteria
Spread across Petri Dish Proteus species most evident

Arrangement basis for classification

Monotrichous; 1 flagella Lophotrichous; tuft at one end Amphitrichous; both ends Peritrichous; all around bacteria

Observe Picture in Micro Lab.

Mono- or Lophotrichorus

Pili
Short protein appendages Adhere bacteria to surfaces
E. coli has numerous types
K88, K99, F41, etc.

smaller than flagella

F-pilus; used in conjugation Flotation; increase boyancy

Pellicle (scum on water) More oxygen on surface

Antibodies to will block adherance Exchange of genetic information

F-Pilus for Conjugation

Capsule or Slime Layer

Glycocalyx - Polysaccharide on external surface Adhere bacteria to surface
S. mutans and enamel of teeth

Prevents Phagocytosis
Complement cant penetrate sugars

Cytoplasm
80% Water {20% Salts-Proteins)
Osmotic Shock important

DNA is circular, Haploid

Advantages of 1N DNA over 2N DNA More efficient; grows quicker Mutations allow adaptation to environment quicker

Plasmids; extra circular DNA

Antibiotic Resistance

No organelles (Mitochondria, Golgi, etc.)

Cell Membrane
Bilayer Phospholipid Water can penetrate Flexible Not strong, ruptures easily
Osmotic Pressure created by cytoplasm

Cell Wall
Peptido-glycan Polymer (amino acids + sugars) Unique to bacteria Sugars; NAG & NAM
N-acetylglucosamine N-acetymuramic acid

D form of Amino acids used not L form

Hard to break down D form

Amino acids cross link NAG & NAM

Cell Wall Summary

Determine shape of bacteria Strength prevents osmotic rupture 20-40% of bacteria Unique to bacteria Some antibiotics effect directly
Penicillin

Video Clip on Cell Wall

Teichoic Acids
Gram + only Glycerol, Phosphates, & Ribitol Attachment for Phages

Lipopolysaccharide (LPS)
Endotoxin or Pyrogen
Fever causing Toxin nomenclature
Endo- part of bacteria Exo- excreted into environment

Structure
Lipid A Polysaccharide
O Antigen of E. coli, Salmonella

G- bacteria only
Alcohol/Acetone removes

 Functions

LPS (contd)

Toxic; kills mice, pigs, humans Pyrogen; causes fever

G- septicemia; death due to LPS DPT vaccination always causes fevers

Heat Resistant; hard to remove Detection (all topical & IV products)

Adjuvant; stimulates immunity

Rabbits (measure fever) Horse shoe crab (Amoebocytes Lyse in presence of LPS)

LPS (contd.)
Appearance of Colonies
Mucoid = Smooth (lots of LPS or capsule) Dry = Rough (little LPS or capsule)

O Antigen of Salmonella and E. coli

2,000 different O Ags of Salmonella 100s different O Ags of E. coli
E. coli O157

O Ags differ in Sugars, not Lipid A

 Resistant structure

Endospores

Heat, irradiation, cold Boiling >1 hr still viable

Takes time and energy to make spores Location important in classification

Central, Subterminal, Terminal

Bacillus stearothermophilus -spores

Used for quality control of heat sterilization equipment

Bacillus anthracis - spores

Used in biological warfare

G+ vs. G G+
Thicker cell wall Teichoic Acids

G Endotoxin - LPS

Which are more sensitive to Penicllin? Alcohol/Acetone affects which more?

Prokaryotes vs. Eukaryotes

Cell Wall Teichoic Acids LPS Endospores Circular DNA Plasmids

Eukaryote Cell Structure

Bacteria Cell

Prokaryotes
Cells that do not have a nucleus Exist almost every where on earth Grow in numbers so great you can see them with the unaided eye Are placed in either the Eubacteria or the Archebacteria Kingdoms Make up the smaller of the two kingdoms

Eubacteria
Make up the larger of the two prokaryote kingdoms Generally are surrounded by a cell wall composed of complex carbohydrates

Cyanobacteria
Photosynthetic bacterium Bluish-greenish color Contain membranes that carry out the process of photosynthesis Do not contain the same type of chloroplasts as plants do This bluish-greenish algae can be found nearly everywhere on earth. Can survive in extremely hot environments and even extremely cold environment

Archaebacteria
Lack important carbohydrate found in cell walls Have different lipids in their cell membrane Different types of ribosomes Very different gene sequences Archaebacteria can live in extremely harsh environments They do not require oxygen and can live in extremely salty environments as well as extremely hot environments.

Identifying Prokaryotes
Cell Shape Cell Wall Movement

Bacterium Shapes
Cocci~ Sphere shaped bacteria Bacillus~ Rod shaped bacteria Spirrillium ~ Spiral shaped bacteria Flagella~ Leg-like structures that help to propel the bacterium.

Gram + and Gram Bacterium Cell Walls

Cellular Walls
Chemical nature of a cell wall can be determined by Gram Staining By finding out what color the cell produces when it is gram stained you can figure out the type of carbohydrates in the cell wall

Movement
Flagella ~ Tail like structure the whips around to propel the bacterium Cillia ~ Miniature flagella surround the cell that help to swim Non motile ~ Sticky cillia like structures that keep the bacterium from moving

Flagella

Bacteria and their energy

Autotrophs Chemotrophs Heterotrophs

Autotrophs
Make their own energy Using Solar energy Eg. Cyanobacteria

Chemotrophs
Make own Energy Using Chemical energy Eg. Archaebacteria

Heterotrophs
Obtain food By eating Eg. E-coli

Bacteria Respiration
Obligate Anaerobes Facultative Anaerobes Obligate Aerobes Live without Oxygen Can live with or without oxygen Cannot live without oxygen.

Bacteria Reproduction
Binary Fission Conjugation Spore Formation

Cellular organism copies its genetic information then splits into two identical daughter cells

Conjugation
A type of Bacteria Sex Two organism swap genetic information, that contains the information such as a resistance to penicillin

Spore Formation: Endospore

A type of dormant cell Exhibit no signs of life Highly resistant to environmental stresses such as: -High temperatures -Irradiation -Strong acids -Disinfectants Endospores are formed by vegetative cells in response to environmental signals that indicate a limiting factor for vegetative growth, such as exhaustion of an essential nutrient.

Symbiosis
Close relationship between to species in which at least one species benefits from the other Live together for LIFE

Parasitism
Bacteria exploit the host cell, injuring them Eg. Mychobacterium tuberculosis

Mutualism
Symbiosis in which two of the species live together in such a way that both benefit from the relationship Eg. E-coli

Nitrogen Fixations
Process by which nitrogen in the atmosphere is converted into a form that can be used by living things