Team:Paris Bettencourt/Project/Eliminate Smell

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BACKGROUND What if you could smell just like yourself? Sweat is initially odorless but bacteria present on your sking process these compounds and release stinky sulphure volatiles.

AIMS Find the bacteria responsible for body odor in human samples Develop CRISPRs that target the bacteria responsible for body malodor to find a natural odorless strain Formulate a probiotic deodorant cream that contains natural odorless bacteria to cure body odor

RESULTS Lorem ipsum dolor sit amet, consectetur adipiscing elit. Cras elementum mauris interdum lacus venenatis aliquet. In ac nulla semper, convallis orci non, vestibulum nisi. Maecenas quis ultricies tellus. Sed vulputate tempor ultrices.

BIOBRICKS AgaA gene responsible for body odor in pSB1C3.

Smell test:

Figure 2:14 people smelled two tubes of E. coli grown to saturation in LB. One culture carried synthetic agaA and other an empty vector control. 13 people out of 14 rated the E. coli carrying agaA as more smelly (pink) than the control(violet). Experiments were double-blind. Significance was confirmed by Chi square test (p-value = 0.001341).

GC:
We performed a GC analysis in the E. coli expressing agaA and used E. coli transformed with an empty vector as a control. COMPOUND X was detected in the E. coli expressing agaA .

(GC FIGURE)

Corynebacterium striatum was found in natural sweat samples. Via 16s sequencing, we found different Corynebacterium species:

(PHILOGENETIC TREE)

Additionally, we conducted a smell test on these samples.

(FIGURE)

Figure 1:Enzymes responsible for body odor in the human axile (Tauch (2013). Daily battle against body odor: towards the activity of the axillary microbiota. Trends in Microbiology 21(6):305–312).

1) Microbiome study: looking for genes responsible for body odor

There are several enzymes responsible for body odor: agaA, aecD, L-Ldh, D-Ldh, Ldh, and AckA. We Sanger-sequenced human axilary sweat samples for these genes.
(MÉGANE'S FIGURE)
(MÉGANE'S RESULTS: XXX genes are responisble for body odor).

The AgaA enzyme of Coryneacterium striatum is a major source of "pungent" or "musky" aromatics in human body odor (Acuna G. (2003). A specific bacterial aminoacylase cleaves odorant precursors secreted in the human axilla. J Biol Chem. 278 (8), 5718-27). It hydrolyzes 3-methyl-2-hexenoyl-glutamine (3M2H-gln) into 3-methyl-2-hexenoic acid (3M2H) and free glutamine. The enzyme is known to have a low specificity for the acyl group, and to act on a range of glutamine conjugates. We cloned agaA into the standard BioBrick vector, and expressed it in E. coli.

agaA was successfully cloned into E. coli.

A noticeable odor was produced by agaA-expressing E. coli grown in selective LB, described variously as "beer-like" or "cheese-like". We took this to be evidence that the enzyme was functional and acting on a non-native substrate in LB media. We confirmed this observation with a formal smell text (Figure 3) and GC analysis (Figure 4).

2) CRISPRs: finding natural odorless mutants

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3) Probiotic cream: a cure for body odor

(Figure for the cream.)

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