Difference between revisions of "Part:BBa K5115034"
 
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<partinfo>BBa_K5115034 short</partinfo>
 
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<html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/5115/czh/mineral-logo.svg" alt="contributed by Fudan iGEM 2024"></html>
 
__TOC__
 
__TOC__
 
===Introduction===
 
===Introduction===
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The csoS operon, originating from the ''Halothiobacillus neapolitanus'', 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). In literature, α-carboxysomes have been extensively studied and successfully utilized in ''Escherichia coli'' for enhancing carbon fixation efficiency and optimizing metabolic pathways. The csoS operon includes key structural proteins including csoS4B, csoS1C, csoS1A, csoS1B, csoS1D, csoS4A, and CsoS2, which play crucial roles in forming the shell and encapsulating cargo enzymes, including those required for hydrogen production. The operon serves as a model for synthetic biology applications, particularly in constructing nanoreactors capable of enhancing catalytic functions through encapsulation of heterologous enzymes. The successful expression of this operon in ''E. coli'' demonstrates its potential for industrial and biotechnological applications, enabling the creation of efficient microbial systems for sustainable bioprocessing.<ref>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. https://doi.org/10.1038/s41467-020-19280-0.</ref>
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The csoS operon, originating from the ''Halothiobacillus neapolitanus'', 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)<ref>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. https://doi.org/10.1038/s41594-020-0387-7.</ref>. In literature, α-carboxysomes have been extensively studied and successfully utilized in ''Escherichia coli'' for enhancing carbon fixation efficiency and optimizing metabolic pathways. The csoS operon includes key structural proteins including csoS4B, csoS1C, csoS1A, csoS1B, csoS1D, csoS4A, and CsoS2, which play crucial roles in forming the shell and encapsulating cargo enzymes, including those required for hydrogen production. The operon serves as a model for synthetic biology applications, particularly in constructing reactors capable of enhancing catalytic functions through encapsulation of heterologous enzymes. The successful expression of this operon in ''E. coli'' demonstrates its potential for industrial and biotechnological applications, enabling the creation of efficient microbial systems for sustainable bioprocessing<ref>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. https://doi.org/10.1038/s41467-020-19280-0.</ref>.
  
 
===Usage and Biology===
 
===Usage and Biology===
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The csoS operon composite part is designed for use in ''E. coli'' to facilitate the expression and assembly of α-carboxysomes. These microcompartments are advantageous for engineering metabolic pathways, especially in enhancing the efficiency of carbon fixation and enzyme activity. The operon includes genes that encode shell proteins, such as CsoS1A and CsoS1B, which form the structural framework of the carboxysome. Additionally, the CsoS2 protein is vital for organizing Rubisco within the carboxysome, playing a role analogous to that of CcmM in β-carboxysomes, where it links cargo and shell assemblies.
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In our experiment, we don't need to use the very original csoS operon. We choose to remove the csoS3 subunit from this operon. For more details about the part we eventually choose to adopt, please check [https://parts.igem.org/Part:BBa_K5115065 BBa_K5115065(cso, without csoS3)]
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Research indicates that the CsoS2 protein exists in two isoforms—CsoS2A and CsoS2B—differing in size and functionality, with the longer form being particularly important for the proper assembly of the empty α-carboxysome shells. Experimental results have shown that deletion of CsoS2 results in the failure to form shell structures in E. coli, underscoring its necessity in carboxysome assembly. The C-terminus of CsoS2 has been identified as an encapsulation peptide (EP) that facilitates the incorporation of cargo enzymes into the shell, allowing for the construction of functional nanoreactors.
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By employing this composite part in ''E. coli'', researchers can harness the natural assembly mechanisms of carboxysomes to create efficient systems for producing biofuels and other valuable bioproducts. The incorporation of enzymes such as [FeFe]-hydrogenase into the α-carboxysome shell offers a promising strategy for enhancing hydrogen production while protecting these sensitive enzymes from oxygen, thereby optimizing catalytic activities within a controlled microenvironment.
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===Characterization===
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===Sequence and Features===
  
 
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Latest revision as of 10:15, 2 October 2024


csoS operon

contributed by Fudan iGEM 2024

Introduction

The csoS operon, originating from the Halothiobacillus neapolitanus, 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)[1]. In literature, α-carboxysomes have been extensively studied and successfully utilized in Escherichia coli for enhancing carbon fixation efficiency and optimizing metabolic pathways. The csoS operon includes key structural proteins including csoS4B, csoS1C, csoS1A, csoS1B, csoS1D, csoS4A, and CsoS2, which play crucial roles in forming the shell and encapsulating cargo enzymes, including those required for hydrogen production. The operon serves as a model for synthetic biology applications, particularly in constructing reactors capable of enhancing catalytic functions through encapsulation of heterologous enzymes. The successful expression of this operon in E. coli demonstrates its potential for industrial and biotechnological applications, enabling the creation of efficient microbial systems for sustainable bioprocessing[2].

Usage and Biology

In our experiment, we don't need to use the very original csoS operon. We choose to remove the csoS3 subunit from this operon. For more details about the part we eventually choose to adopt, please check BBa_K5115065(cso, without csoS3)

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 133
    Illegal NotI site found at 6599
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 291
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2805
    Illegal AgeI site found at 799
    Illegal AgeI site found at 1750
    Illegal AgeI site found at 2431
    Illegal AgeI site found at 4933
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 191


References

  1. ↑ 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. https://doi.org/10.1038/s41594-020-0387-7.
  2. ↑ 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. https://doi.org/10.1038/s41467-020-19280-0.