Contribution

Streptomyces bacteria are responsible for various antibiotics available in the market. Much of the potential for potential drugs is hidden because the genes producing these compounds belong to cryptic clusters. In an effort to increase the production of these compounds, we have developed parts to optimize the expression of the genes responsible for their synthesis, using the cosmomycin produced by Streptomyces olindensis as a proof of concept.

Sections on SARP

The cosmomycin SARP, Streptomyces Antibiotic Regulatory Protein (BBa_K4590001), is responsible for regulating the genes within the cluster that synthesizes cosmomycin in the genome of Streptomyces olindensis. (Example of a SARP [1]).

Sections on Efflux Pumps

The ABC Transporter ATP Binding Protein (BBa_K4590002) and the ABC Transporter Membrane Protein (BBa_K4590003) constitute an ABC transporter (BBa_K4590004), responsible for exporting cosmomycin to the extracellular environment. Reducing the concentration of the secondary metabolite inside the microorganism would limit negative feedback and inhibit the production of more compound, functioning as a resistance mechanism [2].

In addition to cosmomycin export, other mechanisms of resistance to the toxicity of secondary metabolites include glutathione peroxidase (BBa_K4590005) and UvrA-like protein (BBa_K4590006), which act as antioxidants for the free radicals generated by anthracyclines and repair DNA damage, respectively. Both are derived from the genome of Streptomyces olindensis[3].

The composite part (BBa_K4590007) represents the combination of all these strategies to protect against the toxicity caused by the production of the secondary metabolite.

Sections on Malonate Incorporation

Streptomyces olindensis does not have a documented malonate transporter, based on a previous study [4]. Therefore, we proposed using the di-Tricarboxylate Transporter (BBa_K4590008) from the genome of Streptomyces hygroscopicus, a similar organism, to incorporate malonate from the extracellular medium, to be used as a precursor for the secondary metabolite. To handle the increased malonate inside the cell, we included Malonyl-CoA Synthase (BBa_K4590009), which will be used in the synthesis of anthracyclines. Together, they form the part Di-Tricarboxylate Transporter and Malonyl-CoA Synthase (BBa_K4590010).

We also included a Plasmid backbone of malonate production (BBa_K4590011) for assembling a Malonate incorporation vector (BBa_K4590012).

Sections on scientific divulgation

Furthermore, our team has also developed a comprehensive synthetic biology handbook that serves as a fundamental introduction guide for new teams who starting their journey in synthetic biology and iGEM. In terms of scientific development, encompassing both dry lab" and "wet lab" aspects, as well as a valuable contribution to future iGEM teams and society at large. This handbook has been designed to provide a solid starting point for those looking to embark on the journey of forming an iGEM team and adquire more knowledge in synthetic biology, and was first shared at an educational event at the Universidade de Brasília (UnB).

The handbook covers a wide range of essential topics, including metabolic engineering, genetic analysis, primer design, biological circuit construction, structural modeling of proteins, molecular docking, and others. It represents a valuable tool for skill development in synthetic biology, enabling new teams to understand and apply complex concepts in an accessible manner.