Introduction

1. Why do we detect benzo[a]pyrene?

   Because when food,such as fried food ,which is heated above 300 degrees celcius,food will release benzo[a]pyrene recycled oil usually contain benzo[a]pyrene[1].

2. The harm of benzo[a]pyrene

   Pathway:from skin,breathing in,eating Carcinogenic,environmental pollution, Skin irritation,eye irritation[1].

3. Taiwanese regulation
   
4. National regulation
   • Europe:The same to Taiwanese regulation[2]
   • American:The MCL has been set at 0.2 ppb[3]

Circuit Design

So far,we don’t find the protein which can combine with the Benzo[a]pyrene,so first we must degradate it by Laccase. And its product is Bap-1,6-quinone which can combine with the protein,in order that we can detect Benzo[a]pyrene.[4] We put the gene fragment which can produce the Laccase in the E.coli. Let it produce itself.

QsrR is a protein which can combine with Bap-1,6-quinone.[5]As Laccase,we put the gene fragment in the E.coli. Let it produce itself.

We design a gene part make QsrR repress sfRFP’s production. Usually,QsrR is on the Binding Site,and red fluorescent protein will not be produced. When QsrR combine with Bap-1,6-quinone, QsrR will be activated and go away. And the transcription will carry on,then it will produce red fluorescent protein.

Result

1. Whether benzo[a]pyrene can enter e.coli or not
   1. Method
   2. Result
   3. Discussion
2. Whether e.coli is alive in the poisons, condition or not
   1. Method
   2. Results
   3. Discussion
3. The relation between the concentration of benzo[a]pyrene and illumination of RFP
   1. Method
   2. Results
   3. Discussion

Reference

• [1] 黄靖芬，李来好，陈胜军，杨贤庆..烟熏食品中苯并(a)芘的产生机理及防止方法smoked foods Mechanism of benzo (a) pyrene and prevention methods
• [2] 降低食品中多環芳香族碳氫化合物含量之作業指引(草案) reduce operating guidelines in food polycyclic aromatic hydrocarbon content of the (draft)
• [3] Basic Information about Benzo(a)pyrene in Drinking Water
• [4] Hadibarata T, Kristanti RA. Identification of metabolites from benzo[a]pyrene oxidation by ligninolytic enzymes of Polyporus sp. S133. J Environ Manage. 2012 11/30;111(0):115-9.(2013 IGEM CUHK)
• [5] Ji Q, Zhang L, Jones MB, Sun F, Deng X, Liang H, et al. Molecular mechanism of quinone signaling mediated through S-quinonization of a YodB family repressor QsrR. Proceedings of the National Academy of Sciences. 2013 March 26;110(13):5010-5. (2013 IGEM CUHK)