Difference between revisions of "Team:UCSC/Safety"
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| − | <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B<sub>12</sub> we sought out an FDA approved microbe; however, we resorted to using a non-toxic cyanobacteria as a model for | + | <p><span class="firstword">When</span> choosing an organism to help us produce acetaminophen and vitamin B<sub>12</sub> we sought out an FDA-approved microbe; however, we resorted to using a non-toxic cyanobacteria, <i>Synechococcus elongatus</i> strain PCC 7942, as a model for <i>Arthrospira platensis</i>. <i>A. platensis</i> is a generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.</p> |
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| − | <p>We are growing two different strains of <i>S. elongatus</i>: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to <i>A. platensis</i>, we plan to use PCC 7002, which requires external B<sub>12</sub> for survival, as a growth reference for the detection of vitamin B<sub>12</sub>. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.</p> | + | <!--<p>We are growing two different strains of <i>S. elongatus</i>: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to <i>A. platensis</i>, we plan to use PCC 7002, which requires external B<sub>12</sub> for survival, as a growth reference for the detection of vitamin B<sub>12</sub>. PCC 7942 was obtained from the University of California, San Diego (UCSD) iGEM team and PCC 7002 was obtained from Dr. Donald Bryant at Pennsylvania State University.</p>--> |
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<h3>VITAMIN B<sub>12</sub></h3> | <h3>VITAMIN B<sub>12</sub></h3> | ||
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| − | The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH<sub>2</sub>). FMNH<sub>2</sub> is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the <i>ssuE</i> gene [19,20]. | + | The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH<sub>2</sub>). FMNH<sub>2</sub> is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the <i>ssuE</i> gene<sup>[19,20]</sup>. |
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<h3><b>T O X I C I T Y</b> TESTS</h3> | <h3><b>T O X I C I T Y</b> TESTS</h3> | ||
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| − | <p>In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.</p> | + | <p>In order to judge whether our project would possibly be safe for consumption, we performed toxicity tests for the acetaminophen pathway in PCC 7942. Varying amounts of acetaminophen were added to liquid culture and the OD of each sample was observed. Liquid culture samples containing 1.32 mM, 6.62 mM, and 13.23 mM of acetaminophen were left to oxidize overnight and the color change was compared with liquid cultures samples containing similar concentrations of 4-aminophenol.</p> --> |
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Latest revision as of 23:58, 1 November 2017
SAFETY
When choosing an organism to help us produce acetaminophen and vitamin B12 we sought out an FDA-approved microbe; however, we resorted to using a non-toxic cyanobacteria, Synechococcus elongatus strain PCC 7942, as a model for Arthrospira platensis. A. platensis is a generally recognized as safe organism and our genes were chosen from bio-safety level 1 (BSL-1) organisms.
L A B SAFETY
Proper PPE for BSL-1 organisms are worn in our laboratory. Within the lab space, every person is expected to be wearing gloves, long pants, close-toed shoes, a lab coat, and protective goggles. All personnel also must be accompanied with one other person so that there is no one alone in lab.
P R O J E C T SAFETY
VITAMIN B12
G E N E S U S E D:
I. ssuE --> produces FMN reductase
Organism: Synechoccocous elongatus strain PCC 7002
Biosafety Level I
The synthesis of 5,6-DMB first requires the presence of reduced flavin mononucleotide (FMNH2). FMNH2 is reduced from flavin mononucleotide (FMN) by FMN reductase, which is encoded with the ssuE gene[19,20].
FMNH2 + NADP+ = FMN + NADPH
II. bluB --> 5,6-DMB synthase
Organism: Sinorhizobium meliloti 1021
Biosafety Level I
ACETAMINOPHEN
G E N E S U S E D:
I. 4ABH --> 4-aminophenol
Organism: Agaricus bisporus
Biosafety Level I
II. nhoA --> converts 4-aminophenol into acetaminophen
Organism: Escherichia coli
Biosafety Level I
