Team:Paris Bettencourt/Project/Eliminate Smell
From 2014.igem.org
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
|
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. |
Part1 | Part2 | Part3 | Part4 | Part5 |
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)
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).
Motivation
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Ut imperdiet diam eget quam imperdiet imperdiet. Mauris dapibus risus felis, sed ornare diam accumsan aliquet. Sed eu turpis porta, porttitor tortor et, condimentum augue. Curabitur a maximus nisi. Vivamus vitae magna ex. Donec congue auctor odio vitae tempus. In a gravida neque, et tristique tortor. Phasellus a odio sit amet enim ornare lobortis. Morbi sodales, diam non rutrum aliquam, ligula mauris consectetur urna, sed interdum quam risus sit amet enim. Aenean euismod enim magna, id pretium eros molestie non. Proin rutrum lobortis leo, sit amet congue erat. Nulla congue pellentesque augue porta dignissim. Pellentesque quis ex sollicitudin, condimentum risus varius, aliquet ipsum. Ut pulvinar aliquet maximus. Praesent imperdiet interdum commodo.
Aims
Instead of killing these bacteria -such as deodorants do-, we want to find a a natural strain that does not have agaA in it, and create a probiotic cream.
In order to do so, the 'Don't sweat it team' has followed these steps:
1)agaA BioBrick: Create a E. coli strain that contains agaA
2)CRISPRs selection: Select a Corynebacterium strain that has a mutant agaA gene.
3) Natural strains: find a natural strain that contains a mutant agaA or no agaA at all. We will use samples taken from volunteers and perform deep sequencing and do bacteria identification and isolation.
4) Probiotic cream: Design a product that can be used as a probiotic deodorant using a natural Corynebacterium strain that does not release sulphur compounds.
Results
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Ut imperdiet diam eget quam imperdiet imperdiet. Mauris dapibus risus felis, sed ornare diam accumsan aliquet. Sed eu turpis porta, porttitor tortor et, condimentum augue. Curabitur a maximus nisi. Vivamus vitae magna ex. Donec congue auctor odio vitae tempus. In a gravida neque, et tristique tortor. Phasellus a odio sit amet enim ornare lobortis. Morbi sodales, diam non rutrum aliquam, ligula mauris consectetur urna, sed interdum quam risus sit amet enim. Aenean euismod enim magna, id pretium eros molestie non. Proin rutrum lobortis leo, sit amet congue erat. Nulla congue pellentesque augue porta dignissim. Pellentesque quis ex sollicitudin, condimentum risus varius, aliquet ipsum. Ut pulvinar aliquet maximus. Praesent imperdiet interdum commodo.