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 as a model for Arthrospira platensis. A. platensis is 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

We are growing two different strains of S. elongatus: 7002 and 7942. While PCC 7942 will act as our host organism due to its metabolic similarity to A. platensis, we plan to use PCC 7002, which requires external B₁₂ for survival, as a growth reference for the detection of vitamin B₁₂. 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.

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