Microbial Cell Organisation ‘Traditionally, cellular organisms have been divided into two broad categories, based on their cell type. They are either prokaryotic or eukaryotic, In general, prokaryotes are smaller, simpler, with a lot less stuff, while eukaryotes are larger, more complex, and more cluttered. The crux of their key difference can be deduced from their names “Pro” means “before,” while means “true,” indicating that prokaryotes lack a true nucleus while eukaryotes have a true nucleus. More recently, microbiologists have been against the term prokaryote because it lumps both bacteria and the more recently discovered Archaea in the same category. Both cells are prokaryotic because they lack a nucleus and other cell structures (such as mitochondria, Golgi apparatus, endoplasmic reticulum, etc), but they aren’t closely related genetically. So, to honor these differences they refer to the groups as the archaea, the bacteria, and the eukaryotes.. Prokaryotic Cells Organisation A prokaryote is a simple, unicellular organism that lacks an organized nucleus or other ‘membrane-bound organelle. © Prokaryotes lack an organized nucleus and other membrane-bound organelles. + Prokaryotic DNA is found in a central part of the cell called the nucleoid. © The cell wall of a prokaryote acts as an extra layer of protection, helps maintain cell shape, and prevents dehydration. '* Prokaryotic cell size ranges from 0.1 to 5.0 pm in diameter. * The small size of prokaryotes allows quick entry and diffusion of ions and molecules to other parts of the cell while also allowing fast removal of waste products out of the cell Cell Morphology Cell morphology is a reference to the shape of a cell. The shape dictates how that cell will grow, reproduce, obtain nutrients, move, and it’s important to the cell to maintain that shape to function properly. Cell morphology can be used as a characteristic to assist in identifying particular microbes but it’s important to note that cells with the same morphology are not necessarily related. Bacteria tend to display the most representative cell morphologies, with the most common examples listed below:Diversity of Morphology in Prokaryotes Coceus os C Spiritum wi Stak Hypha ‘Budcing and appendaged bacteria Cocci (Spheres) Arrangement: - Diplococci: pairs - Chains (e.g. streptococci) - Clusters (e.g. Staphylococci) in a square) ina cube) Bacilli - Rods - Coccobacilli (very short rods) - Filamentous (long rods) - Vibrios (curved rods) ~ Blunt or Square ends Arrangements: cells arranged singly, in short chains, palisade or Chinese Lettering Less Common Shapes: + Spirilla rigid helices - Spirochetes - flexible helices (usually quite long, internalized flagella) - Coryneform - variable appearance (usually rod-like) - Pleomorphie - variable, irregular shapes, occasionally branched - Flattened rectangles, triangles & trapezoids (rare) Less Common Shapes: - Spirilla - rigid helices - Spirochetes - flexible helices (usually quite long, internalized flagella) - Coryneform - variable appearance (usually rod-like) - Pleomorphic - variable, irregular shapes, occasionally branched - Flattened rectangles, triangles & trapezoids (rare)Cell Size Cell size, just like cell morphology, is not a trivial matter to a cell. There are reasons why most archaeal/bacterial cells are much smaller than eukaryotic cells. Much of it has to do with the advantages derived from being small. These advantages relate back to the surface-to-volume ratio of the cell, a ratio of the external cellular layer in contact with the environment compared to the liquid inside. This ratio changes as a cell increases in size. Let us look at a 2 im cell in comparison with a cell that is twice as large at 4 jum. r=2pm ‘7 surface area = 50.3 um2 r= 1m wolume = 335 jms \surface area = 12.6 ym2 = volume = 4.2 m3 The surface-to-volume ratio of the smaller cell is 3, while the surface-to-volume ratio of the larger cell decreases to 1.5. Think of the cell surface as the ability of the cell to bring in nutrients and let out waste products. The larger the surface area, the more possibilities exist for engaging in these activities. Based on this, the larger cell would have an advantage. Now think of the volume as representing what the cell has to support. As the surface-to-volume ratio goes down, it indicates that the cell has less of an opportunity to bring in the nutrients needed to support the cell’s activities — activities such as growth and reproduction. So, small cells grow and reproduce faster. This also means that they evolve faster over time, giving them more opportunities to adapt to environments. Microbial Cell Components All cellular components consist of macromolecules such as proteins, polysaccharides, phospholipids, DNA, RNA, etc. These macromolecules are made up of subunits (primary structure) of sugars, amino acids and nucleotides. The subunits are in turn made up of macronutrients such as C, H, O, N, S and P, etc. The arrangement/sequence of the subunits in the macromolecule determines the properties and the function of these macromolecules. For example the amino acid sequence in a protein determines the properties and functions of that protein, same is with the other macromolecules. The immense diversity of life is as a result of the difference within the primary structure of biological macromolecules. Macromolecules that make up Cell Components Macromolecule Primary subunits | Cell components Proteins ‘Amino acids Plasma membrane, cytoplasm, Ribosomes, Flagella, etc. Polysaccharides Sugar Cell wall, capsules, inclusions Phospholipids Fatty acids Membranes Nucleic acids Nucleotide Nucleoid, plasmids, ribosomes Microbial Cell structure Examination of the internal structure of cells reveals two patterns, called prokaryote and eukaryote. Prokaryotes include the Bacteria and the Archaea and consist of small and structurallyrather simple cells, Eukaryotes are typically much larger than prokaryotes and contain an assortment of membrane enclosed cytoplasmic structures, These include, most prominently, the DNA-containing nucleus but also mitochondria and chloroplasts, organelles that specialize in supplying the cell with energy, and_—_various Structure of Prokaryotic and Eukaryotic Cells Bacteria cell structures and their functions Cell structure | Function Plasma Selectively permeable barrier, mechanical boundary membrane _| of cell, nutrient and waste transport, location of many metabolic processes (respiration, photosynthesis), detection of environmental cues for chemotaxis ‘Gas vacuole | Buoyaney for floating in aquatic environments Ribosomes | Protein synthesis Inclusion ‘Storage of earbon, phosphate, and other substances bodies ‘Nucleoid ‘Localization of genetic material (DNA) Periplasmic | Contains hydrolytic enzymes and binding proteins for nutrient processing and uptake Cell wall ‘Provides shape and protection from osmotic sess Capsules and | Resistance to phagocytosis, adherence to surfaces slime layers Fimbriac and | Attachment to surfaces, bacterial mating [pili [ Flagella ‘Swimming motility ‘Endospore | Survival under harsh environmental conditions other organelles.Structure of the Cell Components All cells (bacterial, archaeal, eukaryotic) share four common components: Cytoplasm, Deoxyribonucleic acid, Ribosomes, Cytoplasmic Membrane Cytoplasm — The cytoplasm, the inside of the cell, is a gel-like aqueous mixture of macromolecules—proteins, lipids, nucleic acids, and polysaccharides—small organic molecules (mainly precursors of macromolecules), various inorganic ions, and ribosomes. It provides an aqueous environment for the chemical reactions that take place in a cell. The cytoplasm of all cells is. separated by a permeability barrier called the cytoplasmic membrane from the outside Deoxyribonucleic acid or DNA is the genetic material of the cell, the instructions for the cell’s abilities and characteristics. This complete set of genes, referred to as a genome, is localized in an iregularly-shaped region known as the nucleoid in bacterial and archaeal cells, and enclosed into a membrane-bound nucleus in eukaryotic cells. Ribosomes —. Composed of both RNA and protein, ribosomes are the work benches (factories) of protein synthesis of the cell. ‘There are some distinct differences between the ones found in bacteria/archaea and the ones found in eukaryotes, in terms of size and location. The ribosomes of bacteria and archaea are found floating in the cytoplasm, while many of the eukaryotic ribosomes are organized along the endoplasmic reticulum, a eukaryotic organelle. Ribosomes are measured using the Svedberg unit, which corresponds to the rate of sedimentation when centrifuged. Bacterial/archaeal ribosomes have a measurement of 70S as a sedimentation value, while eukaryotic ribosomes have a measurement of 80S, an indication of both their larger size and mass. Cytoplasmic Membrane ~ one of the outer boundaries of every cell is the plasma membrane or cell membrane. (A plasma membrane can be found elsewhere as well, such as the membrane that bounds the eukaryotic nucleus, while the term cell membrane refers specifically to this boundary of the cell proper). The plasma membrane separates the cell's inner contents from the surrounding environment. While not a strong layer, the plasma membrane participates in several crucial processes for the cell, particularly for bacteria and archaea, which typically only have the one membrane: © * Acts as a semi-permeable barrier to allow for the entrance and exit of select molecules. It functions to let in nutrients, excrete waste products, and possibly keep out dangerous substances such as toxins or antibiotics. * Performs metabolic processes by participating in the conversion of light or chemical energy into a readily useable form known as ATP. This energy conservation involves the development of a proton motive force (PMF), based on the separation of charges across the membrane, much like a battery. * “Communicates” with the environment by binding or taking in small molecules that act as signals and provide information important to the cell. The information might relate to nutrients or toxin in the area, as well as information about other organismsStructure of the Plasma Membrane When talking about the details of the plasma membrane it gets a little bit complicated, since bacteria and eukaryotes share the same basic structure, while archaea have marked differences. The plasma membrane is often described by the fluid-mosaic model, which accounts for the movement of various components within the membrane itself. The general structure is explained by the separation of individual substances based on their attraction or repulsion of water. The membrane is typically composed of two layers (a bilayer) of phospholipids, which form the basic structure, Each phospholipid is composed of a polar region that is hydrophilie (“water loving”) and a non-polar region that is hydrophobie (“water fearing”). The phospholipids will spontaneously assemble in such a way as to keep the polar regions in contact with the aqueous environment outside of the cell and the cytoplasm inside, while the non-polar regions are sequestered in the middle, much like the jelly in a sandwich, The phospholipids themselves are composed of a negatively-charged polar head which is a phosphate group, connected by a glycerol linkage to two fatty acid tails. The phosphate group is hydrophilic while the fatty acid tails are hydrophobic. While the membrane is not considered to be particularly strong, it is strengthened somewhat by the presence of additional lipid ‘components, stich as the steroids in eukaryotes and the sterol-like hopanoids in bacteria. Embedded and associated with the phospholipid bilayer are various proteins, with myriad functions, Proteins that are embedded within the bilayer itself are called integral proteins while proteins that associate on the outside of the membrane are called peripheral proteins. Some of the peripheral proteins are anchored to the membrane via a lipid tail, and many associate with specific integral proteins to fulfill cellular functions. Integral proteins are the dominant type, representing about 70-80% of the proteins associated with a plasma membrane, while the peripheral proteins represent the remaining 20-30%. J Ula Baayer Integra Protein Peripheral Protein Plasma Membrane Structure.‘The membrane contains various enzymes involved in respiration, and metabolism and in synthesis of capsular and cell wall component. Bacterial Cell Walls About 90% of bacteria have a cell wall and they typically have one of two types: a gram positive cell wall or a gram negative cell wall. The two different cell wall types can be identified in the lab by a differential stain known as the Gram stain. After the invention of the electron microscope in the 1940s, it was found that the staining difference correlated with differences in their cell walls. After this stain technique is applied the gram positive bacteria will purple, while the gram negative bacteria will stain pink, Overview of Bacterial Cell Walls The cell wall is found outside of the cell membrane. It’s an additional layer that typically provides some strength that the cell membrane lacks, by having a semi-rigid structure. Both gram positive and gram negative cell walls contain an ingredient known as peptidoglycan (also known as murein). This particular substance hasn't been found anywhere else on Earth, other than the cell walls of bacteria. But both bacterial cell wall types contain additional ingredients as well, making the bacterial cell wall a complex structure overall, particularly when compared with the cell walls of eukaryotic microbes. The cell walls of eukaryotic microbes are typically composed of a single ingredient, like the cellulose found in algal cell walls or the chitin in fungal cell walls. The bacterial cell wall performs several functions as well, in addition to providing overall strength to the cell. It also helps maintain the cell shape, which is important for how the cell will grow, reproduce, obtain nutrients, and move. It protects the cell from osmotic lysis, as the cell moves from one environment to another or transports in nutrients from its surroundings. Since water can freely move across both the cell membrane and the cell wall, the cell is at risk for an osmotic imbalance, which could put pressure on the relatively weak plasma membrane. Studies have actually shown that the internal pressure of a cell is similar to the pressure found inside a fully inflated car tire. That is a lot of pressure for the plasma membrane to withstand! The cell wall can keep out certain molecules, such as toxins, particularly for gram negativebacteria. And lastly, the bacterial cell wall can contribute to the pathogenicity or disease — causing ability of the cell for certain bacterial pathogens. Structure of Peptidoglyean The peptidoglycan, is the main ingredient that both bacterial cell walls have in common. Peptidoglycan is a polysaccharide made of two glucose derivatives, N-acetylglucosamine (NAG) and N-acetyimuramie acid (NAM), alternated in long chains, The chains are cross- linked to one another by a tetrapeptide that extends off the NAM sugar unit, allowing a lattice- like structure to form. The four amino acids that compose the tetrapeptide are: L-alanine, D- glutamine, L-lysine or meso-diaminopimelic acid (DPA), and D-alanine. Typically only the L-isomeric form of amino acids are utilized by cells but the use of the mirror image D-amino acids provides protection from proteases that might compromise the integrity of the cell wall by attacking the peptidoglycan. The tetrapeptides can be directly cross-linked to one another, with the D-alanine on one tetrapeptide binding to the L-lysine) DPA on another tetrapeptide. In many gram positive bacteria there is a cross-bridge of five amino acids such as glycine (peptide interbridge) that serves to connect one tetrapeptide to another. In either case the cross-linking serves to increase the strength of the overall structure, with more strength derived from complete cross-linking, where every tetrapeptide is bound in some way to a tetrapeptide on another NAG-NAM chain, Neacetylglucosamine (NAG) Nacetylmuramic acid (NAM) GH Podtutamine Lipsine Tetrapeptide Cross-Linking Dealanine Structure of the PeptidoglycanGram Positive Cell walls The cell walls of gram positive bacteria are composed predominantly of peptidoglycan. In fact, peptidoglycan represents up to 90% of the cell wall, with layer after layer forming around the cell membrane. The NAM tetrapeptides are typically cross-linked with a peptide interbridge and complete cross-linking is common, All of this combines together to create an incredibly strong cell wall. The additional component in a gram positive cell wall is teichoic acid, a glycopolymer, which is embedded within the peptidoglycan layers. Teichoic acids can either be covalently linked to peptidoglycan referred to as wall teichoie acids (WTA) or connected to the cell membrane via a lipid anchor, in which case it is referred to as lipoteichoic acid. TTeichoie Acid e@ eee @ @ ¢ }@ |Pertcoarvcan 3 Peptide Coll membrane Cytorlasm ——_Prowohina Structure of the Cell Wall of Gram Positive Bacteria Teichoic acid play several important roles for the cell, such as; © generation of the net negative charge of the cell, which is essential for development of a proton motive force. © contributes to the overall rigidity of the cell wall, which is important for the maintenance of the cell shape, particularly in rod-shaped organisms. © resistance to adverse conditions such as high temperatures and high salt concentrations, as well as to B-lactam antibiotic Gram Negative Cell Walls The cell walls of gram negative bacteria are more complex than that of gram positive bacteria, with more ingredients overall. They do contain peptidoglycan as well, although only a couple of layers, representing 5-10% of the total cell wall. What is most notable about the gram negativecell wall is the presence of a plasma membrane located outside of the peptidoglycan layers, known as the outer membrane. This makes up the bulk of the gram negative cell wall. The outer membrane is composed of a lipid bilayer, very similar in composition to the cell membrane with polar heads, fatty acid tails, and integral proteins. It differs from the cell membrane by the presence of large molecules known as lipopolysaecharide (LPS), which are anchored into the outer membrane and project from the cell into the environment. Lipopolysaccharide (O-Polysaccharide Porin Protein Core Polysaccharide 6 ¢ Lipid A |_outer Membrane | @ 9 Piel — a : so3g-— Pepidootycan Coll Membrane: Structure of the Cell Wall of Gram Negative Bacteria LPS is made up of three different components: 1) the O-antigen or O-polysaccharide, which represents the outermost part of the structure , 2) the core polysaccharide, and 3) lipid A, which anchors the LPS into the outer membrane. LP’ known to serve many different functions for the cell, such as ‘* contributing to the net negative charge for the cell, © helping to stabilize the outer membrane, and * providing protection from certain chemical substances by physically blocking access to other parts of the cell wall. In addition, LPS plays a role in the © host response to pathogenic gram negative bacteria, The O-antigen triggers an immune response in an infected host, causing the generation of antibodies specific to that part of LPS (think of E. coli 0157). © Lipid A acts as a toxin, specifically an endotoxin, causing general symptoms of illness such as fever and diarrhea.Unusual and Wall-less Bacteria Having emphasized the important of a cell wall and the ingredient peptidoglycan to both the gram positive and the gram negative bacteria, it is equally important to point out a few exceptions as well. Bacteria belonging to the phylum Chlamydiae appear to lack peptidoglycan, although their cell walls have a gram negative structure in all other regards (ie. outer membrane, LPS, porin, ete). It has been suggested that they might be using a protein layer that functions in much the same way as peptidoglycan. This has an advantage to the cell in providing resistance to B-lactam antibiotics (such as penicillin), which attack peptidoglycan. Bacteria belonging to the phylum Tenericutes lack a cell wall altogether, which makes them extremely susceptible to osmotic changes. They often strengthen their cell membrane somewhat by the addition of sterols, a substance usually associated with eukaryotic cell membranes, Many members of this phylum are pathogens, choosing to hide out within the protective environment of ahost.