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<h2 class="content-1" id="titleI" style="color:#FFFF82">II. The Life History of Moth</h2> | <h2 class="content-1" id="titleI" style="color:#FFFF82">II. The Life History of Moth</h2> | ||
<div class="modelingPartContent" id="partI"> | <div class="modelingPartContent" id="partI"> | ||
− | <p class="content">After understanding the life history of the moths from Dr. Huang in Taiwan Agriculture Research Institute | + | <p class="content">After understanding the life history of the moths from Dr. Huang in Taiwan Agriculture Research Institute, we can know that the time of pupa becoming moth ranges from six days to 14 days. So the moths we caught in 20 days must be the larvae in our farm right now. Therefore, we utilized the data of the accumulated number of moth in the next 20 days as the target of our prediction system, and we can use the output to know the time that requires spraying Pantide for prevention.</p> |
</div> | </div> | ||
</div> | </div> | ||
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<h2 class="content-1" id="titleL" style="color:#FFFF82">I. Summary</h2> | <h2 class="content-1" id="titleL" style="color:#FFFF82">I. Summary</h2> | ||
<div class="modelingPartContent" id="partL"> | <div class="modelingPartContent" id="partL"> | ||
− | <p class="content"></p> | + | <p class="content">The aim of this modeling was to predict and simulate the degradation rate of Pantide. Practically, the results would be integrated into our device to promote automatic control system. Once Pantide degraded below the effective level, it will spray the solution to replenish.</p> |
+ | <p class="content">After discussion and experimental verification, we show that UV radiolytic oxidation contributes to the degradation of Pantide. We tried to build a model for simulating the relationship between applied UV light and degradation rate and then combined with UV intensity sensor to calculate the period of spraying Pantide.</p> | ||
</div> | </div> | ||
</div> | </div> | ||
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<h2 class="content-1" id="titleM" style="color:#FFFF82">II. Pantide degradation process</h2> | <h2 class="content-1" id="titleM" style="color:#FFFF82">II. Pantide degradation process</h2> | ||
<div class="modelingPartContent" id="partM"> | <div class="modelingPartContent" id="partM"> | ||
− | <p class="content"></p> | + | <p class="content">For the proteins used as Pantide, the inhibitor cystine knot (ICK) is significant to their function. Pantide with several disulfide bonds is often more stable in solution. If “native protein” is denatured, it loses disulfide bonds and becomes a less stable form, normally called “linear protein,” which is easily degradable.</p> |
+ | <p class="content">Recent research shows that the spider toxin proteins containing ICK structure, for example, ω-hexatoxin-Hv1a (Hv1a), have high stability against temperature, pH, solvents and protease. In contrast, when Hv1a is denatured to linear form, it loses its stability and then degrades rapidly. [1]</p> | ||
+ | <p class="content">There are many possible processes of Pantide degradation we discussed below. (Figure 1) Pantide may have a chance to be reduced to a linear form by reductants or reductases. For both native form and linear form proteins, it may suffer hydrolysis and proteolysis, resulting in denaturing or amino acid cleavage. Also, UV light of sun also leads to Pantide degradation. Though the energy of UV light may not be not enough to break the covalent bonds efficiently, proteins still could undergo radiolytic oxidation.</p> | ||
+ | <div> | ||
+ | <img src=”” class=”picture”> | ||
+ | <p class=”Figure 1. Pantide degradation process”></p> | ||
+ | </div> | ||
+ | <p class="content">According to this degradation process, we attempted to build up our model. The degradation rate of Pantide is contributed by hydrolysis, proteolysis, UV radiolytic oxidation and the reduction to linear form. We considered that the rate Pantide transformed to linear form is a part of degradation rate because linear form protein loses its function and it is also likely to be degraded.</p> | ||
+ | <p class="content">However, since the whole process is too complex to verify, we divided the experiment into three parts, hydrolysis test, proteolysis test and UV radiolytic oxidation test, and further summarized the results to conclude. The purpose is to find out the degradation rate of Pantide and to verify the less stable linear form protein has.</p> | ||
+ | <p class="content">On the other hand, because ICK structure domain mainly contributes the stability of these proteins; we could assume that the degradation processes of three target proteins are roughly the same, for the reason that, we chose Hv1a and Hv1a with lectin (Hv1a-lectin) to demonstrate.</p> | ||
</div> | </div> | ||
</div> | </div> | ||
+ | |||
+ | |||
<!--3--> | <!--3--> | ||
<div class="modelingPart"> | <div class="modelingPart"> | ||
<h2 class="content-1" id="titleN" style="color:#FFFF82">III. Hydrolysis test</h2> | <h2 class="content-1" id="titleN" style="color:#FFFF82">III. Hydrolysis test</h2> | ||
<div class="modelingPartContent" id="partN"> | <div class="modelingPartContent" id="partN"> | ||
+ | <p class="content-1" style="color:#00E600"> i. Theory</p> | ||
+ | <p class="content">Like the reverse direction of polypeptide formation, proteins can be hydrolyzed into their constituent amino acids. The mechanism undergoes an E2 elimination since a nucleotide attach to sp2 hybridized acyl carbon (amide). [2] Therefore, the reaction rate depends on the concentration of both nucleotides and the protein: </p> | ||
+ | <div> | ||
+ | <img src=”” class=”picture”> | ||
+ | <p class=”(1)”></p> | ||
+ | </div> | ||
+ | <p class="content">Where, </p> | ||
+ | <p class="content">[<i>P</i>] is the concentation of Pantide, <i>K<sub>h</sub></i> is the reaction constant of hydrolysis, and <i>k<sub>A</sub></i>, <i>k<sub>N</sub></i>, <i>k<sub>B</sub></i> are the three contributing components of overall hydorlysis which are dependent to condition.</p> | ||
+ | <p class="content"> For Pantide, we would apply the protein solution in the farm at a constant pH. Thus, the rate of hydrolysis is proportional to the concentration of Pantide by a constant <i>K<sub>h</sub></i>, and the concentration of Pantide undergoes an exponential decay as time goes on.</p> | ||
+ | <p class="content-1" style="color:#00E600"> ii. Experimental proof</p> | ||
+ | <p class="content">We tested the chemical stability of both native and linear types of Hv1a and Hv1a-lectin in neutral PBS solvent (phosphate buffered saline, pH=7.4) in 4 ℃ for one day and seven days. SDS-PAGE showed the remained protein concentration and calculated by software ImageJ. (Figure 2, Figure 3)</p> | ||
+ | <p class="content"></p> | ||
<p class="content"></p> | <p class="content"></p> | ||
</div> | </div> |
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