Difference between revisions of "Part:BBa K3051421"
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<partinfo>BBa_K3051421 short</partinfo> | <partinfo>BBa_K3051421 short</partinfo> | ||
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+ | <h2>Comparative Enzyme Activity Assay</h2> | ||
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− | <div align="center"><img height="85%" width="85%" src="https://2019.igem.org/wiki/images/ | + | <div align="center"><img height="85%" width="85%" src="https://2019.igem.org/wiki/images/8/83/T--Warwick--2019-EnzymeLipaseGraph.png"></img></div> |
− | <p><b>Figure 1. | + | <p><b>Figure 1. Relative Comparaison Lipase Activity Assay.</b> Candida Antarctica Lipase A, Bacillus Subtilis Lipase and Compost Lipase enzyme activity were examined using p-Nitrophenol octanoate as a substrate.</p> |
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− | + | Figure 1 compares enzyme activity of Candida Antarctica Lipase A <bbpart>BBa_K3051001</bbpart>, Bacillus Subtilis Lipase <bbpart>BBa_K3051005</bbpart> and Compost Lipase <bbpart>BBa_K3051005</bbpart>. The lipase parts were tested and characterized using a [https://2019.igem.org/Team:Warwick/Results p nitrophenol octanoate assay]. The enzyme activity was determined by measuring change in light absorbance at a wavelength of 400nm, which directly correlated to the concentration of p-Nitrophenol (pNP), caused by lipolysis of p-Nitrophenol octanoate (pNPO) substrate. The data from the pNPO lipolysis assay defined the enzyme activity. | |
− | The enzyme showed 77% identity to Thermostable Lipase A <bbpart>BBa_K258006</bbpart>, and likely conserves a similar function. The conserved domains include a hydrolase site and an RTX toxin related Ca2+ binding site - which could either cause sensitivity to Calcium ions or Calcium ions could act as a cofactor. | + | The lipase was found in a metagenomics search of a compost performed by the University of Gottingen. The enzyme showed 77% identity to Thermostable Lipase A <bbpart>BBa_K258006</bbpart>, and likely conserves a similar function. The conserved domains include a hydrolase site and an RTX toxin related Ca2+ binding site - which could either cause sensitivity to Calcium ions or Calcium ions could act as a cofactor. |
Revision as of 03:25, 22 October 2019
Compost Lipase
Comparative Enzyme Activity Assay
Figure 1. Relative Comparaison Lipase Activity Assay. Candida Antarctica Lipase A, Bacillus Subtilis Lipase and Compost Lipase enzyme activity were examined using p-Nitrophenol octanoate as a substrate.
Figure 1 compares enzyme activity of Candida Antarctica Lipase A BBa_K3051001, Bacillus Subtilis Lipase BBa_K3051005 and Compost Lipase BBa_K3051005. The lipase parts were tested and characterized using a p nitrophenol octanoate assay. The enzyme activity was determined by measuring change in light absorbance at a wavelength of 400nm, which directly correlated to the concentration of p-Nitrophenol (pNP), caused by lipolysis of p-Nitrophenol octanoate (pNPO) substrate. The data from the pNPO lipolysis assay defined the enzyme activity.
The lipase was found in a metagenomics search of a compost performed by the University of Gottingen. The enzyme showed 77% identity to Thermostable Lipase A BBa_K258006, and likely conserves a similar function. The conserved domains include a hydrolase site and an RTX toxin related Ca2+ binding site - which could either cause sensitivity to Calcium ions or Calcium ions could act as a cofactor.
Figure 2. CLMP 3D Structure. The protein has a kDa of 64.6 and an estimated molar extinction coefficient of 75290 M-1cm-1 (calculated using ProtoPram ExPaSy tool); the following 3D structure (calculated using Phyre 2).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]