Description: We intend to design, construct and introduce a synthetic Vitamin B12 exporter (“Synporter”) into a production organism. Thereby, we aim for facilitated and higher yields in the industrial Vitamin B12 production without requiring cell lysis. The B12 Synporter consists of a signal for a Twin Arginine Transporter (Tat) mediated export that is linked to a B12-binding domain. This construct will be expressed in S. typhimurium TA100, R. planticola and S. blattae. We will not only test these different production organisms but also different B12-binding domains.Vitamin B12 is involved in metabolic functions in all organisms, and is therefore an essential nutrient. However, it can only be synthesized by some bacteria and archaea, thus, animals have to obtain it through their diet. Thereby, only animal products like meat and dairy contain B12 in general. This B12 was synthesized by microorganisms colonizing the gastrointestinal tract of those animals and accumulated in the animals’ tissues. Vitamin B12 is one of the most essential biochemicals in the world, and its synthesis is extraordinarily complex. Since the chemical production of Vitamin B12 requires 70 synthesis steps, it is far too technically challenging and expensive. Therefore, its production is facilitated by genetically engineered microorganisms. These are able to produce Vitamin B12 in industrial amounts and achieve a high product quality. However, the produced Vitamin B12 is harvested by cell lysis, which prevents a continuous production. The efficiency of production could be increased by exporting Vitamin B12 outside the cells. To date, a natural cellular Vitamin B12 exporter is unknown.An adult needs approximately 3.0 µg Vitamin B12 per day which is essential for certain functions: it is involved in processes concerning synthesis of DNA, hormones and neurotransmitters and is involved in the formation of the nervous system and blood. Hence, a B12 deficiency can cause diverse diseases like cancer, dementia, depression, pernicious anemia and polyneuropathy.Nowadays, many people do not eat meat or animals products at all. Therefore, these people have a high risk to suffer from B12 deficiency if their diet does not provide enough B12.Vitamin B12 is also used in the industry as it is needed in the biotechnical production of various organic substances. Furthermore, it is added to diverse daily products like toothpaste, fruit gum, non-diary milk or cleaning solution for contact lenses.When we have successfully expressed our constructs in our productions organisms, we will try to maximize the Vitamin B12 yield, and possibly find ways to utilize the periplasmic space to facilitate cytotoxic B12 dependent reactions.
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Year: 2016Visit Wiki
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Updated at: 8/9/16