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Discuss how a recombinant enzyme can be expressed, purified and analyzed.

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Section Problem to be addressed Metabolism – role of fructose-1,6-bisphosphatase DNA Technology – how a gene for an enzyme can be isolated, cloned and placed into a plasmid vector. Proteins – how a recombinant enzyme can be expressed, purified and analysed. Metabolism – how these energy supplies are maintained by metabolic processes. Context Fructose-1,6-bisphosphatase deficiency is inherited in an autosomal recessive manner. Inherited mutations in the FBP1 gene cause a deficiency of the enzyme. The deficient enzyme contains a point mutation at position 122 where an aspartic acid residue is mutated to alanine. Fructose-1,6-bisphosphatase (FBPase). This course work will look at how this enzyme can be cloned, purified and studied. Research on metabolic reprogramming in cancer is progressing at an unprecedented pace. FBPase, is usually found to be downregulated in many cancers and is treated as a tumour-suppressor gene. FBPase, has also been found to play critical roles in tumour initiation and progression in several cancer types and loss of FBPase facilitates the Warburg effect in cancer. FBP inhibitors are also studied as drug candidates for the treatment of diabetes type 2 In a biological setting, the enzyme also plays a key role in hibernation, specifically in bats which requires strict regulation of metabolic processes to facilitate entry into hibernation, maintenance, arousal from hibernation, and adjustments to allow long-term dormancy. The temperature has been shown to decrease the Km of this enzyme slows down the metabolisms in this setting and the way in which glucose is processed.

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