Microorganisms such as cyanobacteria can trap the energy in sunlight through the process of photosynthesis and store it in the chemical bonds of carbohydrate molecules. The principal carbohydrate formed in photosynthesis is glucose. Other types of microorganisms such as nonphotosynthetic bacteria, fungi, and protozoa are unable to perform this process. Therefore, these organisms must rely upon preformed carbohydrates in the environment to obtain the energy necessary for their metabolic processes. Cellular respiration is the process by which microorganisms obtain the energy available in carbohydrates. They take the carbohydrates into their cytoplasm, and through a complex series of metabolic processes, they break down the carbohydrate and release the energy. The energy is generally not needed immediately, so it is used to combine ADP with phosphate ions to form ATP molecules. During the process of cellular respiration, carbon dioxide is given off as a waste product. This carbon dioxide can be used by photosynthesizing cells to form new carbohydrates. Also in the process of cellular respiration, oxygen gas is required to serve as an acceptor of electrons. This oxygen gas is identical to the oxygen gas given off in photosynthesis. The overall mechanism of cellular respiration involves four subdivisions: glycolysis, in which glucose molecules are broken down to form pyruvic acid molecules; the Krebs cycle, in which pyruvic acid is further broken down and the energy in its molecule is used to form high-energy compounds such as NADH; the electron transport system, in which electrons are transported along a series of coenzymes and cytochromes and the energy in the electrons is released; and chemiosmosis, in which the energy given off by electrons is used to pump protons across a membrane and provide the energy for ATP synthesis. Glycolysis. The process of glycolysis is a multistep metabolic pathway that occurs in the cytoplasm of microbial cells and the cells of other organisms. At least six enzymes operate in the metabolic pathway. In the first and third steps of the pathway, ATP is used to energize the molecules. Thus, two molecules of ATP must be expended in the process. Further along in the process, the six-carbon glucose molecule is converted into intermediary compounds and then is split into two three-carbon compounds. The latter undergo additional conversions and eventually form pyruvic acid at the conclusion of the process. During the latter stages of glycolysis, four ATP molecules are synthesized using the energy given off during the chemical reactions. Thus, four ATP molecules are synthesized and two ATP molecules are inserted into the process for a net gain of two ATP molecules in glycolysis. Also during glycolysis, another of the reactions yields enough energy to convert NAD toNADH. The reduced coenzyme (NADH) will later be used in the electron transport system, and its energy will be released. During glycolysis, two NADH molecules are produced. As glycolysis does not use oxygen, the process is considered to be anaerobic. For certain anaerobic organisms, such as certain bacteria and fermentation yeasts, glycolysis is the sole source of energy. It is a somewhat inefficient process because much of the cellular energy remains in the two molecules of pyruvic acid. The Krebs cycle. Following glycolysis, the mechanism of cellular respiration then involves another multistep process called the Krebs cycle, also called the citric acid cycle and the tricarboxylic acid cycle. The Krebs cycle uses the two molecules of pyruvic acid formed in glycolysis and yields high-energy molecules of NADH and FADH and some ATP and carbon dioxide (Figure 1 ). Figure 1 An overview of the processes of cellular respiration showing the major pathways and the places where ATP is synthesized. The Krebs cycle occurs at the cell membrane of bacterial cells and in the mitochondriaof eukaryotic cells. Each of these sausage-shaped organelles of eukaryotic microorganisms possesses inner and outer membranes, and therefore an inner and outer compartment. The inner membrane is folded over itself many times; the folds are called cristae. Along the cristae are the important enzymes necessary for the proton pump and for ATP production. Prior to entering the Krebs cycle, the pyruvic acid molecules are processed. Each three-carbon molecule of pyruvic acid undergoes conversion to a substance called acetyl-coenzyme A, or acetyl-CoA. In the process, the pyruvic acid molecule is broken down by an enzyme, one carbon atom is released in the form of carbon dioxide, and the remaining two carbon atoms are combined with a coenzyme called coenzyme A. This combination forms acetyl-CoA. In the process, electrons and a hydrogen ion are transferred to NAD to form high-energy NADH. Acetyl-CoA now enters the Krebs cycle by combining with a four-carbon acid called oxaloacetic acid. The combination forms the six-carbon acid called citric acid. Citric acid undergoes a series of enzyme-catalyzed conversions. The conversions, which involve up to 10 chemical reactions, are all brought about by enzymes. In many of the steps, high-energy electrons are released to NAD. The NAD molecule also acquires a hydrogen ion and becomes NADH. In one of the steps, FAD serves as the electron acceptor, and it acquires two hydrogen ions to become FADH2. Also, in one of the reactions, enough energy is released to synthesize a molecule of ATP. Since there are two pyruvic acid molecules entering the system, two ATP molecules are formed. Also during the Krebs cycle, the two carbon atoms of acetyl-CoA are released and each forms a carbon dioxide molecule. Thus, for each acetyl-CoA entering the cycle, two carbon dioxide molecules are formed. Since two acetyl-CoA molecules enter the cycle, and each has two carbon atoms, four carbon dioxide molecules will form. Add these four molecules to the two carbon dioxide molecules formed in the conversion of pyruvic acid to acetyl-CoA, and the total is six carbon dioxide molecules. These six CO2molecules are given off as waste gas in the Krebs cycle. They represent the six carbons of glucose that originally entered the process of glycolysis. At the end of the Krebs cycle, the final product formed is oxalo-acetic acid, identical to the oxaloacetic acid which begins the cycle. The molecule is now ready to accept another acetyl-CoA molecule to begin another turn of the cycle. All told, the Krebs cycle forms (per two molecules of pyruvic acid) two ATP molecules, a large number of NADH molecules, and some FADH2 molecules. The NADH and the FADH2 will be used in the electron transport system. The electron transport system. The electron transport system occurs at the bacterial cell membrane and in the cristae of the mitochondria in eukaryotic cells. Here, a series of cytochromes (cell pigments) and coenzymes exist. These cytochromes and coenzymes act as carrier molecules and transfer molecules. They accept high-energy electrons and pass the electrons to the next molecule in the system. At key proton-pumping sites, the energy of the electrons is used to transport protons across the cell membrane or into the outer compartment of the mitochondrion. Each NADH molecule is highly energetic. It accounts for the transfer of six protons across the membrane. Each FADH2 molecule accounts for the transfer of four protons. Electrons pass from NAD to FAD, to other cytochromes and coenzymes, and eventually they lose much of their energy. The final electron acceptor is an oxygen atom. The electron-oxygen combination then takes on two protons to form a molecule of water(H2O). As a final electron receptor, oxygen is responsible for removing electrons from the system. If oxygen were not available, electrons could not be passed among the coenzymes, the energy in electrons could not be released, the proton pump could not be established, and ATP could not be produced. Chemiosmosis. The actual production of ATP in cellular respiration takes place duringchemiosmosis. As previously noted, chemiosmosis involves the pumping of protons through special channels in the membranes of mitochondria from the inner to the outer compartment. In bacteria, the pumping occurs at the cell membrane. The pumping establishes a proton gradient. Once the gradient is established, protons pass down the gradient through molecular particles. In these particles, the energy of the protons is used to generate ATP, using ADP and phosphate ions as the starting points. The energy production in cellular respiration during chemiosmosis is substantial. Most biochemists agree that in prokaryotic microorganisms, a total of 36 molecules of ATP can be produced during cellular respiration. In eukaryotic cells, the number is 34 molecules of ATP. Two molecules of ATP are produced as the net gain of glycolysis, so the grand total is 38 molecules of ATP (36 in eukaryotes). These ATP molecules may then be used in the cell for its needs. Fermentation. Fermentation is an anaerobic process in which energy can be released from glucose even though oxygen is not available. Fermentation occurs in yeast cells, and a form of fermentation takes place in bacteria. In yeast cells, glucose can be metabolized through cellular respiration, as in other cells. When oxygen is lacking, however, glucose is still changed to pyruvic acid via glycolysis. The pyruvic acid is first converted to acetaldehyde and then to ethyl alcohol. The net gain of ATP to the yeast cell is two molecules–-the two molecules of ATP normally produced in glycolysis. Yeasts are able to participate in fermentation because they have the necessary enzyme to convert pyruvic acid to ethyl alcohol. This process is essential because it removes electrons and hydrogen ions from NADH during glycolysis. The effect is to free the NAD so that it can participate in future reactions of glycolysis. Yeasts are therefore used in both bread and alcohol production. Alcohol fermentation is the process that yields beer, wine, and other spirits. The carbon dioxide given off supplements the carbon dioxide given off during the Krebs cycle and causes bread to rise.