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

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<p class="first">Safety</p>
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<p class="first">Engineering</p><br><br>
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<p class="second">All members</p>
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<p class="description">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.
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<p class="description">All of our team members have basic experimental operation exercises and have a basic understanding of the experiment. In addition, all team members attended experimental safety lectures held by our school, we all have certain concepts for laboratory safety.</p>
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</p>
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<p class="second">Wet Lab</p>
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<p class="second">Filtering system</p>
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<p class="description">In the biological experiment part, the methanolic yeast and Escherichia coli were used, and the experimental contents were all in the first level of Biosafety Level l lab. The operation sites were all in the aseptic operation table, and the personnel were in the experiment. Gloves were worn during the period and equipment were all disinfected with alcohol before and after the experiment.</p>
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<p class="description">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.
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<p class="second">Dry Lab</p>
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</p>
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<p class="description">In the dry experimental group, we conducted experiments that adhered to laboratory safety rules (such as wearing gloves, using volatile drugs in the draft cabinet, etc.) and having relevant professional guidance. Before using the instrument, the yeast will be filtered out in the laboratory to prevent the yeast from leaving the laboratory. All reactions in the design of the future production line will be carry out in the fermentation tank. Keep the reaction environment in a sealed state, then filtering out the polymer. By two protective measures, filter and heat, we can prevent any leakage. For the filter, we will be using a 33kDa filter, the yeast will not pass. As for the heater, all products will be heated 110 ° C for 10 minutes.</p>
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<p class="second">Heating system</p>
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<p class="description">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="second"><br><br>Reference</p>
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<p class="description"><ol>
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<li>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). <br>doi:10.1186/s12934-014-0183-3</li>
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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>
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<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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</ol>
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Revision as of 12:29, 5 October 2018

Engineering



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.



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.
  4. 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.
    doi:10.1128/mcb.5.12.3376
  5. 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.
    doi:10.1007/s00253-014-5732-5