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Microbial Chemistry Dr.Mohammad Shakeeb,MD Specialist in clinical pathology/microbiology and immunology INTRODUCTION • chemistry is an important component of a microbiology. • A microbe can be thought of as a “bag” of chemicals that interact with each other in various ways. • The various ways microorganisms function and survive in their environment depend on their chemical makeup. INTRODUCTION • Even the most simple procaryotic cells consist of very large molecules (macromolecules). Deoxyribonucleic acid (DNA). Ribonucleic acid (RNA). Proteins. Lipids. Polysaccharides. INTRODUCTION • These macromolecules can be broken down into smaller units or “building blocks”. monosaccharides (simple sugars). fatty acids. amino acids. nucleotides. • The macromolecules and building blocks found in cells are collectively referred to as biological molecules. INTRODUCTION • building blocks can be broken down into even smaller molecules such as water, carbon dioxide, ammonia, sulfides, and phosphates, which in turn can be broken down into atoms of carbon (C), hydrogen (H), oxygen (O),nitrogen (N), sulfur (S), phosphorus (P). INTRODUCTION • Organic chemistry is the study of compounds that contain carbon. • inorganic chemistry involves all other chemical reactions. • biochemistry is the chemistry of living cells. ORGANIC CHEMISTRY ORGANIC CHEMISTRY • Organic compounds are compounds that contain carbon. • organic chemistry is that branch of the science of chemistry that specializes in the study of organic compounds. • Although some organic compounds are associated with living organisms, many are not. ORGANIC CHEMISTRY Carbon Bonds • Carbon atoms have a valence of four. • carbon atom can bond to four other atoms. ORGANIC CHEMISTRY Carbon Bonds • The uniqueness of carbon lies in the ability of its atoms to bond to each other to form a multitude of compounds. • There are three ways in which carbon atoms can bond to each other: ORGANIC CHEMISTRY Carbon Bonds • When atoms of other elements attach to available bonds of carbon atoms, compounds are formed. • if only hydrogen atoms are bonded to the available bonds, compounds called hydrocarbons are formed. ORGANIC CHEMISTRY Carbon Bonds • When more than two carbons are linked together, longer molecules are formed. ORGANIC CHEMISTRY Carbon Bonds • A series of many carbon atoms bonded together is referred to as a chain. • Long-chain carbon compounds are usually liquids or solids, whereas short-chain carbon compounds are gases. ORGANIC CHEMISTRY Cyclic Compounds • Carbon atoms may link to carbon atoms to close the chain, forming rings or cyclic compounds. • An example is benzene, which has six carbons and six hydrogens. • some compounds contain fused rings (e.g., double- or triple-ringed compounds). ORGANIC CHEMISTRY BIOCHEMISTRY BIOCHEMISTRY • Biochemistry is the study of biology at the molecular level and can, thus, be thought of as the chemistry of life or the chemistry of living organisms. • Biomolecules are usually large molecules (called macromolecules) and include carbohydrates, lipids, proteins, and nucleic acids. BIOCHEMISTRY • Microorganisms also absorb their essential nutrients into the cell by various means. • These nutrients are then used in metabolic reactions as sources of energy and as building blocks for enzymes, structural macromolecules, and genetic materials. BIOCHEMISTRY • Carbohydrates • Carbohydrates are biomolecules composed of carbon, hydrogen, and oxygen, in the ratio of 1:2:1, or simply CH2O. Monosaccharides • The simplest carbohydrates are sugars, and the smallest sugars. • The most important monosaccharide in nature is glucose (C6H12O6). BIOCHEMISTRY BIOCHEMISTRY • Monosaccharides may contain from three to nine carbon atoms. BIOCHEMISTRY Disaccharides • Disaccharides (di meaning “two”) are double-ringed sugars that result from the combination of two monosaccharides. • synthesis of a disaccharide from two monosaccharides by removal of a water molecule is called a dehydration synthesis reaction. • The bond holding the two monosaccharides together is called a glycosidic bond; it is a type of covalent bond. BIOCHEMISTRY BIOCHEMISTRY • Sucrose, lactose, and maltose are examples of disaccharides. • Disaccharides react with water in a process called a hydrolysis reaction, which causes them to break down into two monosaccharides: BIOCHEMISTRY • Peptidoglycan is a complex macromolecular network found in the cell walls of all members of Bacteria. • Peptidoglycan consists of a repeating disaccharide, attached by polypeptides (proteins) to form a lattice that surrounds and protects the entire bacterial cell. • Several antibiotics (including penicillin) prevent the final cross-linking of the rows of disaccharides, thus weakening the cell wall and leading to lysis (bursting) of the bacterial cell. BIOCHEMISTRY • Polysaccharides • Polysaccharides can be defined as carbohydrates that contain many monosaccharides. • Polysaccharides, such as glycogen, starch, and cellulose, are examples of polymers—molecules consisting of many similar subunits. In the case of polysaccharides, the repeating subunits are monosaccharides. BIOCHEMISTRY • Many bacteria produce polysaccharide capsules, which protect the bacteria from being phagocytized (eaten) by white blood cells. • algal cells have cellulose cell walls to provide support and shape as well as protection against the environment. BIOCHEMISTRY • When polysaccharides combine with other chemical groups (amines, lipids, and amino acids), extremely complex macromolecules are formed that serve specific purposes. • Glucosamine and galactosamine (amine derivatives of glucose and galactose, respectively) are important constituents of the supporting polysaccharides in connective tissue fibers, cartilage, and chitin. BIOCHEMISTRY • Chitin is the main component of the hard outer covering of insects, spiders, and crabs, and is also found in the cell walls of fungi. • The main portion of the rigid cell wall of bacteria consists of amino sugars and short polypeptide chains that combine to form the peptidoglycan layer. BIOCHEMISTRY Lipids • Lipids are essential constituents of almost all living cells. • lipids can be classified into the following categories: • Waxes • Fats and oils • Phospholipids • Glycolipids • Steroids • Prostaglandins and leukotrienes BIOCHEMISTRY Fatty Acids • the building blocks of lipids. • Fatty acids are long-chain carboxylic acids that are insoluble in water. • Saturated fatty acids contain only single bonds between the carbon atoms. • Monounsaturated fatty acids have one double bond in the carbon chain. • Polyunsaturated fatty acids contain two or more double bonds. BIOCHEMISTRY Waxes • A wax consists of a saturated fatty acid and a longchain alcohol. • The waxes that are present in the cell walls of Mycobacterium tuberculosis (the causative agent of tuberculosis) are responsible for several interesting characteristics of this bacterium. • M. tuberculosis cell be phagocytized by a phagocytic white blood cell (a phagocyte), the waxes protect the cell from being digested. BIOCHEMISTRY • the waxes in the cell walls of M. tuberculosis make the organism difficult to stain. • In the acid-fast staining procedure, for example, it is necessary to heat the carbolfuchsin dye to drive it into the cell. • once the cell has been stained, the waxes prevent decolorization of the cell when a mixture of acid and alcohol is applied. • Because the cell does not decolorize in the presence of acid, the organism is described as being acid-fast. BIOCHEMISTRY Fats and Oils • Fats and oils are also known as triglycerides, because they are composed of glycerol (a three-carbon alcohol) and three fatty acids. BIOCHEMISTRY Phospholipids • Phospholipids contain glycerol, fatty acids, a phosphate group, and an alcohol. • There are two types: glycerophospholipids (also called phosphoglycerides) and sphingolipids. BIOCHEMISTRY • In addition to phospholipids, the outer membrane of Gram-negative bacterial cell walls contains lipoproteins and lipopolysaccharide (LPS). • LPS consists of a lipid portion and a polysaccharide portion. • The lipid portion is called lipid-A or endotoxin. • When endotoxin is present in the human. bloodstream, it can cause very serious physiologic conditions (e.g., fever and septic shock). • The cell walls of Gram-positive bacteria do not contain LPS. BIOCHEMISTRY Proteins • All proteins are polymers of amino acids. • Some proteins are the structural components of membranes, cells, and tissues. • others are enzymes and hormones that chemically control the metabolic balance within both the cell and the entire organism. • they vary widely in the number of amino acids present and in the sequence of amino acids as well as their size, configuration, and functions. BIOCHEMISTRY • Proteins contain carbon, hydrogen, oxygen, nitrogen, and sometimes sulfur. Amino Acid Structure • A total of 23 different amino acids have been found in proteins, 20 primary or naturally occurring amino acids plus 3 secondary amino acids. • Each amino acid is composed of carbon, hydrogen, oxygen, and nitrogen. • 3 of the amino acids also have sulfur atoms in the molecule. BIOCHEMISTRY BIOCHEMISTRY BIOCHEMISTRY Protein Structure • When water is removed, by dehydration synthesis, amino acids become linked together by a covalent bond, referred to as a peptide bond. BIOCHEMISTRY Enzymes • Enzymes are protein molecules produced by living cells as “instructed” by genes on the chromosomes. • Enzymes are referred to as biological catalysts— biologic molecules that catalyze metabolic reactions. • catalyst is defined as an agent that speeds up a chemical reaction without being consumed in the process. BIOCHEMISTRY Nucleic Acids • DNA is the “hereditary molecule”—the molecule that contains the genes and genetic code. • DNA makes up the major portion of chromosomes. • The information in DNA must flow to the rest of the cell for the cell to function properly. • this flow of information is accomplished by RNA molecules. • RNA molecules participate in the conversion of the genetic code into proteins and other gene products. BIOCHEMISTRY Structure • The building blocks of nucleic acid polymers are called nucleotides. • Nucleotides consist of three subunits: a nitrogencontaining (nitrogenous) base, a five-carbon sugar (pentose), and a phosphate group, joined together. • The building blocks of DNA are called DNA nucleotides, whereas the building blocks of RNA are called RNA nucleotides. BIOCHEMISTRY BIOCHEMISTRY BIOCHEMISTRY • three types of RNA, which are named for the function they serve: messenger RNA (mRNA), ribosomal RNA (rRNA), and transfer RNA (tRNA). BIOCHEMISTRY BIOCHEMISTRY BIOCHEMISTRY • The bonding forces of the double stranded polymer cause it to assume the shape of a double-helix, which is similar to a right-handed spiral staircase. BIOCHEMISTRY DNA Replication • It occurs by separation of the DNA strands and the building of complementary strands by the addition of the correct DNA nucleotides. • The most important enzyme required for DNA replication is DNA polymerase. • Other enzymes are also required, including DNA helicase and DNA topoisomerase (which initiate the separation of the two strands of the DNA molecule), primase (which synthesizes a short RNA primer), and DNA ligase (which connects fragments of newly synthesized DNA). BIOCHEMISTRY • The duplicated DNA of the chromosomes can then be separated during cell division. • so that each daughter cell contains the same number of chromosomes, the same genes, and the same amount of DNA as in the parent cell. BIOCHEMISTRY Gene Expression • It was Francis Crick who, in 1957, proposed what is referred to as the Central Dogma to explain the flow of genetic information within a cell. • DNA → mRNA → protein • The Central Dogma (also known as the “one gene– one protein hypothesis”) states that: 1. The genetic information contained in one gene of a DNA molecule is used to make one molecule of mRNA by a process known as transcription. BIOCHEMISTRY 2. The genetic information in that mRNA molecule is then used to make one protein by a process known as translation. • The primary enzyme involved in transcription is called RNA polymerase (DNA-dependent RNA polymerase). BIOCHEMISTRY • The process by which the genetic information within an mRNA molecule is used to make a specific protein is called translation. • Translation occurs at a ribosome.