Team:UT-Knoxville/Parts
UTK
iGEM
Parts Collection
Our parts set was developed in order serve as an elegant means of creating product libraries from a toxic byproduct of oil refinement, toluene. Our library production module, BBa_K1966003, is created via the combination of BBa_K1966000 and BBa_K1966001. The xylene monooxygenase (XO) activity of BBa_K1966000 converts the CH3 group of toluene and its derivatives into CH2OH. The benzyl alcohol dehydrogenase from Pseudomonas putida, BBa_K1966001, converts a -CH2OH group of benzyl alcohol and its derivatives into -CHOH. These parts are promiscuous toward toluene derivatives and their resultant alcohols, thus allowing a library of aldehydes to be created from various substrates. Though only partially characterized, the regulatory element of our collection, BBa_K1966004, potentially provides control over the expression of the catalytic enzymes noted above. The Pu Promoter allows for control of gene expression through the introduction of toluene and xylR, thus, creating a control system that regulates the expenditure of cellular resources.
Basic Part
The part BBa_K1966001 contains P. putida BADH (benzyl alcohol dehydrogenase). Our project has shown several functions of this enzyme, some of which are novel:
- Conversion of benzyl alcohol to benzaldehyde.
- Conversion of 3-methylbenzyl alcohol to m-tolualdehyde.
- Conversion of 4-methylbenzyl alcohol to p-tolualdehyde.
- Conversion of 3-chlorobenzyl alcohol to 3-chlorobenzaldehyde (novel function).
- Conversion of 3-methoxybenzyl alcohol to m-anisaldehyde (novel function).
- Use of P. putida BADH to enhance biosynthesis of indigo (novel function).
Composite Part
The part BBa_K1966001 contains P. putida BADH (benzyl alcohol dehydrogenase). Our project has shown several functions of this enzyme, some of which are novel:
- Conversion of toluene to benzaldehyde.
- Conversion of m-xylene to m-tolualdehyde.
- Conversion of p-xylene to p-tolualdehyde.
- Enhanced biosynthesis of indigo.
