Escherichia coli Polyphosphate Kinase Enzyme
This part is a protein generator device producing the Polyphosphate Kinase (PPK) gene from Escherichia coli.
Usage and Biology
PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain (Akiyama et al., 1992). This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long) - see illustration below.
The E. coli PPK coding sequence was derived from the pBC29 plasmid, published by Akiyama et al., 1992. iGEM York 2015 obtained pBC29 with Dr Jay Keasling's kind assistance.
Akiyama, Masahiro, E. Crooke, and Arthur Kornberg. "The polyphosphate kinase gene of Escherichia coli. Isolation and sequence of the ppk gene and membrane location of the protein." Journal of Biological Chemistry 267.31 (1992): 22556-22561.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 396
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 1750
- 1000COMPATIBLE WITH RFC
iGEM2019_Nanjing China Experiment
This year our team develops a simple solo medium-copy plasmid-based polyphosphate kinase (PPK1) overexpression strategy for achieving maximum intracellular polyphosphate accumulation, so the data can provide some reference to this part.
To test whether the C. f reundii derivative that wasconstructed on the basis of the solo medium-copy strategy could perform well in uptaking of exogenous Pi from the SMW, we compare DH5a, DH5a-MDPP and CF-MCPP
Ps: SMW means Synthetic municipal wastewater
DH5a-MDPP means dual-plasmid which contain high and medium copy DH5a ppk in DH5a
CF-MCPP means solo medium-copy C. f reundii ATCC8090 ppk in C. f reundii ATCC8090
DH5a-HCPP means solo high-copy DH5a ppk in DH5a
CF-MDPP means dual-plasmid which contain high and medium copy C. f reundii ATCC8090 ppk in C. f reundii ATCC8090
Wang X , Wang X , Hui K , et al. Highly Effective Polyphosphate Synthesis, Phosphate Removal and Concentration Using Engineered
Environmental Bacteria Based on a Simple Solo Medium-copy Plasmid Strategy[J]. Environmental Science & Technology, 2017:acs.est.7b04532.
Kato, J.; Yamada, K.; Muramatsu, A.; Ohtake, H. Genetic improvement of Escherichia coli for enhanced biological removal of phosphate from wastewater. Appl. Environ. Microbiol. 1993, 59 (11),3744−3749.
Jones, K. L.; Kim, S.-W.; Keasling, J. Low-copy plasmids can perform as well as or better than high-copy plasmids for metabolic engineering of bacteria. Metab. Eng. 2000, 2 (4), 328−338.
Liang, M. Z.; Frank, S.; Lunsdorf, H.; Warren, M. J.; Prentice,M. B. Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli. Biotechnol. J.2017, 12 (3),1600415.