Difference between revisions of "Part:BBa K5115067"

Revision as of 09:24, 30 September 2024

mineral, F module

Introduction

This part is made up of Ni-Fe hydrogenase and carboxysome, the former is encapsulated into the latter by EP. All the subunits of the proteins constructed into our ribozyme-assisted polycistronic co-expression system.

The Ni-Fe hydrogenase include the part BBa_K5115020（hox and hyp operon）. Multiple subunits work together to complete the hydrogenase function. Two main subunits are hoxF and hoxH, one working as NADH dehydrogenase, another working as catalytic centre of hydrogen reaction.^[1] The hyp operon plays a critical role in the maturation of hydrogenase. The hypA and hypB are involved in nickel binding and transport, ensuring that the hydrogenase subunits receive the required metal ions for optimal activity.^[2] Through the synergistic integration of the hox and hyp operon, our system effectively enhances hydrogen production and enables the reduction of nickel ions into nanoparticles, thereby maximizing the efficiency of nickel recovery from industrial wastewater.

The carboxysome include the part BBa_K5115065（cso, without csoS3）. It encodes a series of proteins essential for the assembly of α-carboxysomes, a type of microcompartment that facilitates the sequestration and concentration of enzymes involved in carbon fixation, particularly ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco)^[3]These microcompartments are advantageous for engineering metabolic pathways, especially in enhancing the efficiency of carbon fixation and enzyme activity. The overall design not only supports nickel ion reduction but also promotes enhanced carbon capture, thereby contributing to a more sustainable bioprocess.

The EP is the part BBa_K5115002. It is designed to facilitate the effective encapsulation of enzymes within the carboxysome structure, enhancing the efficiency of biochemical reactions. It serves as a linker that connects the target enzymes to the carboxysome, ensuring proper localization and functionality.^[4] In this F module, EP is combined with hoxF. And in U module BBa_K5115066, EP is combined with hoxU. Different sites of combination can influence the effect of the carboxysome encapsulation, so we decided to choose the better one through experient.

Based on the ribozyme-assisted polycistronic co-expression system, all of the proteins above can express at a equalized level. To learn more about our pRAP system, please check part wiki of 2022 Fudan iGEM.

Usage and Biology

The F module harnesses the collaborative power of hydrogenase enzymes, carboxysome compartments, and encapsulation peptides to drive an innovative approach for nickel reduction in E. coli. This integrated module not only advances the biotechnological potential of engineered microorganisms but also addresses environmental concerns related to nickel contamination by converting harmful ions into less toxic nanoparticles.

Characterization

Sequence and Features

References

1. ↑ Löscher, S., Burgdorf, T., Zebger, I., Hildebrandt, P., Dau, H., Friedrich, B., & Haumann, M. (2006). Bias from H2 Cleavage to Production and Coordination Changes at the Ni−Fe Active Site in the NAD+-Reducing Hydrogenase from Ralstonia eutropha. Biochemistry, 45(38), 11658–11665.
2. ↑ Anne K. Jones, Oliver Lenz, Angelika Strack, Thorsten Buhrke, and, & Friedrich*, B. (2004, October 2). NiFe Hydrogenase Active Site Biosynthesis: Identification of Hyp Protein Complexes in Ralstonia eutropha† (world) [Research-article]. ACS Publications; American Chemical Society.
3. ↑ Oltrogge, L. M., Chaijarasphong, T., Chen, A. W., Bolin, E. R., Marqusee, S., & Savage, D. F. (2020). Multivalent interactions between CsoS2 and Rubisco mediate α-carboxysome formation. Nature structural & molecular biology, 27(3), 281–287.
4. ↑ Li, T., Jiang, Q., Huang, J., Aitchison, C. M., Huang, F., Yang, M., Dykes, G. F., He, H. L., Wang, Q., Sprick, R. S., Cooper, A. I., & Liu, L. N. (2020). Reprogramming bacterial protein organelles as a nanoreactor for hydrogen production. Nature communications, 11(1), 5448.