Difference between revisions of "Team:CCU Taiwan/Safety"

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<a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design"><li class="list" id="project2">Design</li></a>
 
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                     <li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a><hr>
 
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<p class="description">&emsp;&emsp;According to the literature [3], heating at 70 °C for 30 minutes is sufficient to kill the cells. We used a temperature of 110 ° C for 10 minutes to kill any cells that might pass through the filter.
 
<p class="description">&emsp;&emsp;According to the literature [3], heating at 70 °C for 30 minutes is sufficient to kill the cells. We used a temperature of 110 ° C for 10 minutes to kill any cells that might pass through the filter.
 
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<p class="description">Following were our heating test:</p>
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<p class="description">Figure 1. P. pastoris heated at 30 °C for 30 minutes</p>
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<p class="description">Figure 2. P. pastoris heated at 50 °C for 30 minutes</p>
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<p class="description">Figure 3. P. pastoris heated at 70 °C for 30 minutes</p>
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<p class="description">&emsp;&emsp;We found that our P. pastoris not survival after were heated at 50 °C for 30 minutes. Thus, our production line heat process would kill P. pastoris passing through the filter.</p>
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<p class="second"><br><br>Reference</p>
 
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<li>Bacher, G., Szymanski, W. W., Kaufman, S. L., Zöllner, P., Blaas, D., & Allmaier, G. (2001). Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses.<br> Journal of Mass Spectrometry, 36(9), 1038–1052. <br>doi:10.1002/jms.208</li>
 
<li>Bacher, G., Szymanski, W. W., Kaufman, S. L., Zöllner, P., Blaas, D., & Allmaier, G. (2001). Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses.<br> Journal of Mass Spectrometry, 36(9), 1038–1052. <br>doi:10.1002/jms.208</li>
 
<li>3. Martínez, D., Menéndez, C., Echemendia, F. M., Hernández, L., Sobrino, A., & Trujillo, L. E. (2015). Kinetics of sucrose hydrolysis by immobilized recombinant Pichia pastoris cells in a batch reactors. J Microb Biochem Technol, 7, 294-6.</li>
 
<li>3. Martínez, D., Menéndez, C., Echemendia, F. M., Hernández, L., Sobrino, A., & Trujillo, L. E. (2015). Kinetics of sucrose hydrolysis by immobilized recombinant Pichia pastoris cells in a batch reactors. J Microb Biochem Technol, 7, 294-6.</li>
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<li>4. Cregg, J. M., Barringer, K. J., Hessler, A. Y., & Madden, K. R. (1985). Pichia pastoris as a host system for transformations. Molecular and Cellular Biology, 5(12), 3376–3385.<br> doi:10.1128/mcb.5.12.3376</li>
 
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<li>5. Ahmad, M., Hirz, M., Pichler, H., & Schwab, H. (2014). Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. Applied Microbiology and Biotechnology, 98(12), 5301–5317.<br> doi:10.1007/s00253-014-5732-5</li>
 
 
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Revision as of 04:12, 15 October 2018

Safety



  Genetically modified organisms escaping from lab is a serious problem since it will bring unpredictable impacts to our ecosystem. In our part design and production line, we were very conscious of biosafety. The yeast will pass the following process to achieve complete destruction. We also dealt cautiously with waste produced from the experiment and participated in safety training to ensure everyone conducting experiments had a minimal probability to cause biohazard pollution.

Filtering system

  The average size of the P. pastoris used is about 4–6 μm [1], and the filter we used is 33 kDa, which is equivalent to the size of tens of nanometers [2], more than sufficient to trap all the yeast.

Heating system

  According to the literature [3], heating at 70 °C for 30 minutes is sufficient to kill the cells. We used a temperature of 110 ° C for 10 minutes to kill any cells that might pass through the filter.

Following were our heating test:

Figure 1. P. pastoris heated at 30 °C for 30 minutes

Figure 2. P. pastoris heated at 50 °C for 30 minutes

Figure 3. P. pastoris heated at 70 °C for 30 minutes

  We found that our P. pastoris not survival after were heated at 50 °C for 30 minutes. Thus, our production line heat process would kill P. pastoris passing through the filter.



Reference

  1. 1. Gmeiner, C., Saadati, A., Maresch, D., Krasteva, S., Frank, M., Altmann, F., … Spadiut, O. (2015). Development of a fed-batch process for a recombinant Pichia pastoris Δoch1 strain expressing a plant peroxidase. Microbial Cell Factories, 14(1).
    doi:10.1186/s12934-014-0183-3
  2. Bacher, G., Szymanski, W. W., Kaufman, S. L., Zöllner, P., Blaas, D., & Allmaier, G. (2001). Charge-reduced nano electrospray ionization combined with differential mobility analysis of peptides, proteins, glycoproteins, noncovalent protein complexes and viruses.
    Journal of Mass Spectrometry, 36(9), 1038–1052.
    doi:10.1002/jms.208
  3. 3. Martínez, D., Menéndez, C., Echemendia, F. M., Hernández, L., Sobrino, A., & Trujillo, L. E. (2015). Kinetics of sucrose hydrolysis by immobilized recombinant Pichia pastoris cells in a batch reactors. J Microb Biochem Technol, 7, 294-6.