Difference between revisions of "Part:BBa K936000"

Revision as of 03:34, 4 October 2012

LC Cutinase

This is the LC Cutinase gene used to break PET into terephthalic acid and ethylene glycol. The enzyme Cutinase is a lypolytic/esterolytic enzyme that degrades cutin, which is found in most plant and fungi' cuticles. They enzyme itself is more commonly found in plants and bacteria. Cutinase also can degrades water soluble esters and insoluble triglycerides. The enzyme hydrolyzes these substrates by creating an acyl enzyme intermediate. A metagenomic analysis was done in order to find more novel enzymes such as lipases, esterases, proteases, and cellulases, as well as furthering our knowledge of protein sequence space in the environment. Naturally, compost samples would have enzymes that degrade cell walls and other compounds with potentially useful applications. A novel homolog of cutinase, known as Leaf Compost Cutinase (LC) was found to have a 57.4% genetic similarity to cutinase found in T. Fusca, meriting an experiment to overproduce the protein in E. Coli. The study found that it's applications could be most valuable in the textile industry, among other material fields.

It should be noted that ethylene glycol is moderately toxic to humans and other animals.

Through p-nitrophenyl butyrate (pNPB) assays the UC Davis team gathered enough data to determine that the LC-Cutinase part (BBa_K936000) most likely exhibits its intended function as an esterase. A detailed description of these assays can be found on the Module Engineering Project page: http://2012.igem.org/Team:UC_Davis/Project/Catalyst.
Additionally, the following data is further described on the UC Davis Cutinase Activity data page: http://2012.igem.org/Team:UC_Davis/Data/Cutinase_Activity

(Above Graph) Results of pNPB assay detailing the constitutive construct as having the highest esterase activity.
(Above Graphs) The graphs above detail higher activity among the constitutive construct and some mutant variants in esterase activity compared to the control and wild type per unit cell.

(Above graph) This graph represents a possible production of cutinase by the cells upon entering stationary phase.

Sequence and Features