Difference between revisions of "Part:BBa K4015005"
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<partinfo>BBa_K4015005 short</partinfo> | <partinfo>BBa_K4015005 short</partinfo> | ||
| − | NusA-Lcp is composed of two basic parts, NusA and | + | NusA-Lcp is composed of two basic parts, NusA and Lcp1VH2. NusA is bonded to Lcp1VH2 to enhance the solubility of Lcp1VH2 and in order to increase its yield. NusA is used to enhance the solubility of Lcp through fusing to binding sites of Lcp. |
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | The solubility of a protein played an important role in the production yields. The previous article brought up a solution, which claims that putting a highly soluble protein to a protein with a low solubility can facilitate protein synthesis. It states that fusing the targeted enzyme to a highly soluble protein can improve the yield of recombinant protein, prevent proteolysis, and increase its solubility. | ||
| + | In our project，our target protein is Lcp1VH2 with a low solubility，an article suggests NusA can act as a fusion partner to improve the production of Lcp1VH2. Therefore, we have inserted NusA as the fusion partner to Lcp, creating a new sequence: NusA-Lcp. Other than increase in solubility led by adding of NusA, we have observed an increase in yield led by NusA. This advantage led by NusA was not expected. Comparing to the original sequence, we have found a significant increase on the productivity of protein once NusA is added, which also benefits the later progression of our project. (image) | ||
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| + | unfortunately, the enzyme activity rate of Nus-Lcp1VH2 did not increase comparing to Lcp1VH2, and even decreased. The process for Lcp to degrade rubber requires oxygen consumption. It utilizes the process of β oxidation to break down bonds within polyisoprene. During β oxidation, Lcp adds two oxygen molecules in between the chemical bonds of polyisoprene. The oxygen consumption rate in the sample tube represents the enzyme activity of Lcp1VH2. | ||
| + | Compared to the sample tube (Supernatant containing Lcp1VH2)，the time for dissolved oxygen dropping to approximately 0mg/l increased from 6 hours to about 16.5 hours in another sample tube (Supernatant containing NusA-Lcp1VH2). This suggests the increase in yield by fusion protein does not increase the enzyme activity. | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K4015005 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4015005 SequenceAndFeatures</partinfo> | ||
Revision as of 10:31, 20 October 2021
NusA-Lcp
NusA-Lcp is composed of two basic parts, NusA and Lcp1VH2. NusA is bonded to Lcp1VH2 to enhance the solubility of Lcp1VH2 and in order to increase its yield. NusA is used to enhance the solubility of Lcp through fusing to binding sites of Lcp.
Usage and Biology
The solubility of a protein played an important role in the production yields. The previous article brought up a solution, which claims that putting a highly soluble protein to a protein with a low solubility can facilitate protein synthesis. It states that fusing the targeted enzyme to a highly soluble protein can improve the yield of recombinant protein, prevent proteolysis, and increase its solubility. In our project，our target protein is Lcp1VH2 with a low solubility，an article suggests NusA can act as a fusion partner to improve the production of Lcp1VH2. Therefore, we have inserted NusA as the fusion partner to Lcp, creating a new sequence: NusA-Lcp. Other than increase in solubility led by adding of NusA, we have observed an increase in yield led by NusA. This advantage led by NusA was not expected. Comparing to the original sequence, we have found a significant increase on the productivity of protein once NusA is added, which also benefits the later progression of our project. (image)
unfortunately, the enzyme activity rate of Nus-Lcp1VH2 did not increase comparing to Lcp1VH2, and even decreased. The process for Lcp to degrade rubber requires oxygen consumption. It utilizes the process of β oxidation to break down bonds within polyisoprene. During β oxidation, Lcp adds two oxygen molecules in between the chemical bonds of polyisoprene. The oxygen consumption rate in the sample tube represents the enzyme activity of Lcp1VH2. Compared to the sample tube (Supernatant containing Lcp1VH2)，the time for dissolved oxygen dropping to approximately 0mg/l increased from 6 hours to about 16.5 hours in another sample tube (Supernatant containing NusA-Lcp1VH2). This suggests the increase in yield by fusion protein does not increase the enzyme activity.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1377
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]