Team:Leiden/Parts

iGEM Leiden Wiki : Parts

Parts

Part	Type	Name	Designer	Size
BBa_K1938000	Coding	pcrA	Sjoerd Seekles, Charlotte van de Velde	2784 bps
BBa_K1938001	Coding	pcrB	Sjoerd Seekles, Charlotte van de Velde	1006 bps
BBa_K1938002	Coding	pcrC	Sjoerd Seekles, Charlotte van de Velde	711 bps
BBa_K1938003	Coding	pcrD	Sjoerd Seekles, Charlotte van de Velde	675 bps
BBa_K1938004	Coding	Chlorite dismutase	Sjoerd Seekles, Charlotte van de Velde	849 bps
BBa_K1938005	Coding	moaA	Valentijn Broeken, Sjoerd Seekles, Charlotte van de Velde	972 bps
BBa_K1938006	Coding	QDH-DHC	Sjoerd Seekles, Charlotte van de Velde	1487 bps
BBa_K1938009	Composite	Chlorite dismutase with Promoter T7 and RBS	Sjoerd Seekles, Charlotte van de Velde	887 bps
BBa_K1938010	Composite	Chlorite dismutase with strong Promoter and strong RBS	Sjoerd Seekles, Charlotte van de Velde	910 bps
BBa_K1938011	Composite	Chlorite dismutase with strong Promoter , medium RBS	Sjoerd Seekles, Charlotte van de Velde	911 bps
BBa_K1938012	Composite	Chlorite dismutase with strong Promoter , weak RBS	Sjoerd Seekles, Charlotte van de Velde	912 bps
BBa_K1938013	Composite	Chlorite dismutase with Constitutive Strong promoter, Strong RBS and TAT signal peptide	Sjoerd Seekles, Charlotte van de Velde	1006 bps

Figure 1. The perchlorate reduction biobrick. This scheme illustrates the genetic composition of our goal: The perchlorate reduction biobrick. Individual genetic elements are elaborated below.

Figure 2. The perchlorate reduction proteins locations in the cell and their function. Molybdenum cofactor (Moco) is built in PcrA and transported via the Tat secretion system into the periplasm together with PcrD, which acts as a chaperone. PcrAB, the catalytic domain of PcrABC, is able to convert perchlorate into chlorate and chlorate into chlorite. Chlorite dismutase (Cld) is able to break chlorite down into chloride and oxygen ions.

PcrA

Perchlorate reductase subunit alpha. ¹ Component of the perchlorate reductase that catalyzes the reduction of perchlorate to chlorite. Is probably also able to reduce chlorate to chlorite. The alpha subunit is likely the catalytic subunit.

PcrB

Perchlorate reductase subunit beta (Perchlorate reductase iron-sulfur subunit). ¹ Component of the perchlorate reductase that catalyzes the reduction of perchlorate to chlorite. The beta subunit may be responsible for electron transfer to the catalytic alpha subunit PcrA. It is probably translocated together with PcrA, which possesses a Tat-type signal.

PcrC

Perchlorate reductase subunit gamma (Perchlorate reductase heme subunit). ¹ Component of the perchlorate reductase that catalyzes the reduction of perchlorate to chlorite. The gamma subunit may be responsible for electron transfer to the beta subunit PcrB.

PcrD

Perchlorate reductase assembly chaperone protein. ¹ May function as a system-specific molybdenum chaperone protein essential for the assembly of the perchlorate reductase PcrAB complex prior to its periplasmic translocation via the Tat pathway.

Chlorite dismutase

Catalyzes the heme-dependent decomposition of chlorite to O2 and chloride with high efficiency and specificity. ² Used to detoxify chlorite, a by-product of the reduction of perchlorate, a primarily anthropogenic pollutant, in perchlorate-respiring bacteria.

MoaA

Catalyzes, together with MoaC, the conversion of 5'-GTP to cyclic pyranopterin monophosphate (cPMP or molybdopterin precursor Z). ²

Qdh-dhc

Quinol dehydrogenase tetra-heme cytochrome and di-heme c-type cytochrome. ²

References

Bender, K. S., Shang, C., Chakraborty, R., Belchik, S. M., Coates, J. D., & Achenbach, L. A. (2005). Identification, characterization, and classification of genes encoding perchlorate reductase. Journal of Bacteriology, 187(15), 5090-5096.
Melnyk, R. A., Clark, I. C., Liao, A., & Coates, J. D. (2014). Transposon and deletion mutagenesis of genes involved in perchlorate reduction in Azospira suillum PS. MBio, 5(1), e00769-13.