Part:BBa_K5143022

A BioGlue secreted by Saccharomyces cerevisiae and attached to the cellulose

Description

Our project aims to create a fully bio-sourced and biodegradable insect trap. To achieve this, the base of our trap will be made of cellulose produced by a bacterium in a co-culture system. The other organism in this co-culture is Saccharomyces cerevisiae, a yeast that will secrete a bioglue composed of spider silk proteins (BBa_K5143001) and barnacle cement proteins (BBa_K5143002). This bioglue will adhere to the cellulose through a Cellulose Binding Domain (CBD) (BBa_K5143008). The adhesion of the bioglue to the cellulose, which forms the base of our trap, will make it sticky, allowing insects to become trapped upon contact.

We selected this bioglue due to its strong adhesive properties as reported in the scientific literature, and all of this is achieved naturally! (See there for more details (BBBa_K5143003)

The BioGlue (Cp19k-MaSp1) is secreted by Saccharomyces cerevisiae, then it binds to the cellulose to functionalize it.

• Saccharomyces cerevisiae is genetically engineered to produce a specialized bioglue, which is a fusion of spider silk proteins and barnacle cement proteins.
• The bioglue is linked to an alpha-factor within the yeast cell, facilitating its secretion into the extracellular environment.
• Once secreted, the bioglue interacts with cellulose through its Cellulose Binding Domain (CBD), effectively binding to the cellulose surface.
• This interaction functionalizes the cellulose, transforming it into a sticky surface that captures insects.
• The CBD ensures strong adhesion between the bioglue and the cellulose, optimizing the trap’s effectiveness.

Construction

The complete sequence of this biobrick was optimized for optimal expression in Saccharomyces cerevisiae and then synthesized. After synthesis, this fragment was combined with another fragment (see yellow fragment) to produce our final plasmid. This integrative plasmid is designed to incorporate the bioglue genes into the yeast genome.

References

1) A bioinspired synthetic fused protein adhesive from barnacle cement and spider dragline for potential biomedical materials - PubMed.
2) Gilbert, C. et al. Living materials with programmable functionalities grown from engineered microbial co-cultures. Nat Mater 20, 691–700 (2021).
3) A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization of Heterologous Protein Secretion and Surface Display in Saccharomyces cerevisiae | ACS Synthetic Biology

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1001
  Illegal PstI site found at 1304
  Illegal PstI site found at 1397
  Illegal PstI site found at 1403
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 51
  Illegal NheI site found at 211
  Illegal PstI site found at 1001
  Illegal PstI site found at 1304
  Illegal PstI site found at 1397
  Illegal PstI site found at 1403
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 832
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1001
  Illegal PstI site found at 1304
  Illegal PstI site found at 1397
  Illegal PstI site found at 1403
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1001
  Illegal PstI site found at 1304
  Illegal PstI site found at 1397
  Illegal PstI site found at 1403
  Illegal AgeI site found at 1552
  
• 1000
  COMPATIBLE WITH RFC[1000]