Difference between revisions of "Team:SDSZ-China"

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<h1> Project Description </h1>
 
<h1> Project Description </h1>
 
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Bear bile, as one of the most famous animal drugs in Traditional Chinese Medicine (TCM), has been recorded in ancient Chinese medicine book as a significant method to treat hepatic and biliary disorders including cancers. Also, its marked pharmacological effects on fever fighting, detoxification and pain reduction have been confirmed through modern investigations. However, the increasing demand of bear bile has caused bears to be in an endangered state: bear poaching and illegal animal trade have greatly dwindled the number of the wild Asiatic black bear. Apart from that, bear bile farming industry has established in Asia to extract bile through “milking” from the bears, which is basically operated through surgically implanting a permanent catheter in the animal’s gall bladder then obtain the drips. It is unquestionable that the bear bile farming process will lead to both physically and psychologically damage in bears.  
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Bear bile, as one of the most famous animal drugs in Traditional Chinese Medicine (TCM), has been recorded in ancient Chinese medicine book as a significant method to treat hepatic and biliary disorders. Also, its marked pharmacological effects on fever fighting, detoxification and pain reduction have been confirmed through modern investigations. However, the increasing demand of bear bile has caused bears to be in an endangered state: bear poaching and illegal animal trade have greatly dwindled the number of the wild Asiatic black bear. Apart from that, bear bile farming industry has established in Asia to extract bile through “milking” from the bears, which is basically operated through surgically implanting a permanent catheter in the animal's gall bladder then obtain the drips. It is unquestionable that the bear bile farming process will lead to both physically and psychologically damage in bears.  
 
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We found that it is possible to convert the main component of goose bile, CDCA, into UDCA, by using two enzymes to catalyst the reaction. The reaction will proceed in two steps. First, CDCA is oxidized by the enzyme 7α-HSDH, coupled by the regeneration of NAD+ with the enzyme LDH, and pyruvate. Similarly, in the second reduction step, the reaction is catalyzed by the enzyme 7β-HSDH, with the reaction that generates NADPH, which takes glucose as reactant and GDH as enzyme.  
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We found that it is possible to convert the main component of goose bile, CDCA, into UDCA, by using two enzymes to catalyst the reaction. The reaction will proceed in two steps. First, CDCA is oxidized in the presence of 7α-HSDH, coupled by the regeneration of NAD+ with the enzyme LDH, and pyruvate. Similarly, in the second reduction step, the enzyme 7β-HSDH catalyze the reaction of 7-oxo-LCA to UDCA with glucose dehydrogenase (GDH) and glucose for the regeneration of cofactor (NADPH) .
 
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Our mission now is to get the four enzymes (7α-HSDH, 7β-HSDH, GDH, and LDH) by expressing their gene in E.coli. After expression, 7α-HSDH and 7β-HSDH will be able to bind to a chitin column because a gene encoding a chitin binding domain is inserted, so no father purification process will be needed. Then we will examine the activity of the enzymes by measuring the rate of NAD+, NADPH production. At last a machine will be made, including the reaction efficiency measuring system, and the enzyme addition controlling system.
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Our mission is to express the four enzymes (7α-HSDH, 7β-HSDH, GDH, and LDH) in the E.coli and test their activities. After expression, all the enzymes will be able to bind to a chitin column because a fragment encoding a chitin binding domain is fused with the target genes. This specific design excels in two specific ways: first, by controlling the  presence of the chitin column in the solution, we can control the process of the reaction. Second, this design simplified the effect of the specific target protein in the extraction of purified bacteria. At last a machine including the reaction efficiency measuring system and the enzyme addition controlling system will be made.
 
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Revision as of 01:54, 1 July 2017


Project Description

Bear bile, as one of the most famous animal drugs in Traditional Chinese Medicine (TCM), has been recorded in ancient Chinese medicine book as a significant method to treat hepatic and biliary disorders. Also, its marked pharmacological effects on fever fighting, detoxification and pain reduction have been confirmed through modern investigations. However, the increasing demand of bear bile has caused bears to be in an endangered state: bear poaching and illegal animal trade have greatly dwindled the number of the wild Asiatic black bear. Apart from that, bear bile farming industry has established in Asia to extract bile through “milking” from the bears, which is basically operated through surgically implanting a permanent catheter in the animal's gall bladder then obtain the drips. It is unquestionable that the bear bile farming process will lead to both physically and psychologically damage in bears.

To find substitutes or alternative of bear bile farming, our team will be working on biological synthesis of the main effective component of this important medicine, UDCA. And this biological method will be more efficient and also cheaper than the original chemical approach, which is used in the current chemical UDCA synthesis industry.

We found that it is possible to convert the main component of goose bile, CDCA, into UDCA, by using two enzymes to catalyst the reaction. The reaction will proceed in two steps. First, CDCA is oxidized in the presence of 7α-HSDH, coupled by the regeneration of NAD+ with the enzyme LDH, and pyruvate. Similarly, in the second reduction step, the enzyme 7β-HSDH catalyze the reaction of 7-oxo-LCA to UDCA with glucose dehydrogenase (GDH) and glucose for the regeneration of cofactor (NADPH) .

Our mission is to express the four enzymes (7α-HSDH, 7β-HSDH, GDH, and LDH) in the E.coli and test their activities. After expression, all the enzymes will be able to bind to a chitin column because a fragment encoding a chitin binding domain is fused with the target genes. This specific design excels in two specific ways: first, by controlling the presence of the chitin column in the solution, we can control the process of the reaction. Second, this design simplified the effect of the specific target protein in the extraction of purified bacteria. At last a machine including the reaction efficiency measuring system and the enzyme addition controlling system will be made.