Uptake

Introduction

There are many kinds of useful substance which we want to use in environment. For example, bitumen contains resin and aromatic hydrocarbons that are used as glues or paved road. Metals which are rare and are thought to increase needs are also useful substance we want to get. However, it needs a large amount of water and energy to pick up usable aromatic compounds and metals. We aim at picking up them using our Eco-tanker. Eco-tanker will run lower energy than existing methods. In this system, we challenged uptake and storage of iron ion in bacterial micro compartment (BMC).

Why is this device needed?

To store iron ion in BMC, we aimed at using iron-binding peptide and applying to construct a BMC fusion protein. We focused on alpha-synuclein, which is natively unfolded protein and composed of 140 amino acids. It is reported that the some metals are bound at the alpha-synuclein and Fe (III) is associated only with the 25 mer C-terminus of alpha-synuclein (MPVDPDNEAYEMPSEEGYQDYEPEA). The Kd value of this peptide was 21 µM evaluated by ITC analysis (1).

What is this device composed inside it?

How does this device work in EcoTanker

Progress

Now we are trying to evaluate the expression and function of iron binding peptide and to ligate the peptide gene to promoter (BBa_B0034) and terminator (BBa_B0016). We also aim at construction of fusion protein containing PduP which is a component of tank protein and the iron-binding peptide, which will enable to store Fe (III) in tank.

References

1. Metal binding to alpha-synuclein peptides and its contribution to toxicity, Brown DR, Biochem Biophys Res Commun (2009)
2. Engineered alpha-synuclein prevents wild type and familial Parkin variant fibril formation, Sode K, et al., Biochem Biophys Res Commun (2005)

Team:Tokyo-NoKoGen/Project/uptake

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