Give the role of two named enzymes in the production of GM organisms. (2 marks)
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Some higher plants, and some animals (termites), have formed associations (symbioses) with diazotrophs. Diazotrophs are microbes. They are intensively studied by microbiologists. Biological nitrogen fixation was discovered by the German agronomist Hermann Hellriegel and Dutch microbiologist Martinus Beijerinck. Biological nitrogen fixation (BNF) occurs when atmospheric nitrogen is converted to ammonia by an enzyme called nitrogenase. Nitrogenases are enzymes used by some organisms to fix atmospheric nitrogen gas (N2). There is only one known family of enzymes that accomplishes this process. All nitrogenases have an iron – and sulfur-containing cofactor that includes a heterometal complex in the active site (e.g., FeMoCo). In most species, this heterometal complex has a central molybdenum atom. However, in some species it is replaced by a vanadium or iron atom. Enzymes responsible for nitrogenase action are very susceptible to destruction by oxygen. Many bacteria cease production of the enzyme in the presence of oxygen. Many nitrogen-fixing organisms exist only in anaerobic conditions, respiring to draw down oxygen levels, or binding the oxygen with proteins. Biological nitrogen fixation (BNF) occurs when atmospheric nitrogen is converted to ammonia by an enzyme called nitrogenase. The reaction for BNF is: N2 + 8 H+ + 8 e− → 2 NH3 + H2. This type of reaction results in N2 gaining electrons (see above equation) and is thus termed a reduction reaction. The exact mechanism of catalysis is unknown due to the technical difficulties biochemists have in actually visualizing this reaction in vitro, so the exact sequence of the steps of this reaction are not completely understood. Despite this, a great deal is known of the process. While the equilibrium formation of ammonia from molecular hydrogen and nitrogen has an overall negative enthalpy of reaction (i.e. it gives off energy), the energy barrier to activation is very high without the assistance of catalysis, which is done by nitrogenases. The enzymatic reduction of N2 to ammonia therefore requires an input of chemical energy, released from ATP hydrolysis, to overcome the activation energy barrier.
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Agricultural plants are one of the most frequently cited examples of genetically modified organisms (GMOs). Some benefits of genetic engineering in agriculture are increased crop yields, reduced costs for food or drug production, reduced need for pesticides, enhanced nutrient composition and food quality, resistance to pests and disease, greater food security, and medical benefits to the world's growing population. Advances have also been made in developing crops that mature faster and tolerate aluminum, boron, salt, drought, frost, and other environmental stressors, allowing plants to grow in conditions where they might not otherwise flourish (Table 1; Takeda & Matsuoka, 2008). Other applications include the production of nonprotein (bioplastic) or nonindustrial (ornamental plant) products. A number of animals have also been genetically engineered to increase yield and decrease susceptibility to disease. For example, salmon have been engineered to grow larger (Figure 1) and mature faster (Table 1), and cattle have been enhanced to exhibit resistance to mad cow disease (United States Department of Energy, 2007).
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