Coding

Part:BBa_K3731000

Designed by: Haoru Song   Group: iGEM21_Nanjing-China   (2021-09-02)
Revision as of 14:25, 6 September 2021 by Hao-Yin (Talk | contribs)


ppk1 in E.Coli BL21

In bacteria, polyP is synthesized by polyphosphate kinase (PPK). PPK consists of PPK1 and PPK2. PPK1 can lengthen the polymer by using the γ-Pi phosphate bond of ATP, and its reversible reaction is to synthesize ATP from ADP and Pi. PPK2 can use polyP to synthesize GTP or ATP reversibly, and mainly synthesize GTP. The ppk1 codes PPK1, which can promote the synthesis(major function) and decomposition(minor function) of polyP with the residue of ATP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


iGEM2020_Nanjing-China Experiment

Group: Nanjing-China 2020
Author: Guangyu Hu

We found that polyP productivity would reach the highest after inoculation for 14 hours and 100ml of synthetic wastewater in a 200ml Erlenmeyer flask has the highest output and the highest conversion efficiency.

PolyP formation is closely associated with ppk1 gene in CPP. This means the cultivation time and volume will affect P consuming a lot. After comparing different cultivation conditions, we found that polyP productivity will reach the highest after inoculation for 14 hours, that is approximately 20mg/L. Besides, 100ml of synthetic wastewater in a 200ml Erlenmeyer flask has the highest output and the highest conversion efficiency. Without bacterial contamination and normal bacterial viability，it can almost convert all inorganic phosphates into polyphosphates.


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.

We test plasmid-borne ppk1 expression in CPP via qRT-PCR analysis, supernatant Pi concentration and optical density of CPP and CWT in Synthetic municipal wastewater(SMW).

Ps: SMW means Synthetic municipal wastewater

CPP means solo medium-copy C. f reundii ATCC8090 ppk in C. f reundii ATCC8090

CWT means wild type C. f reundii ATCC8090

Figure 1）qRT-PCR analysis of plasmid-borne ppk1 expression in CPP

Figure 2）Pi absorption&release

Figure 3）Optical density of CPP grown in SMW compared with CWT

Reference: 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.

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