Biologically produced pigments for tattoo and cosmetic use

This report relates to the project initially discussed in week 1 of HTGAA.

Section 1: Abstract

• Synthetic biology, as a relatively novel field, is full of possibility. It can help us improve efficiency at multiple levels, from gene regulation to bioproduction, or the development of novel molecules of interest.
• Tattoos represent a significant industry, with a revenue of 1.5 bn $ and an approximate yearly increase of 8% since 2007. Despite that, there is a significant lack of regulation on ink composition, which would represent a good way of carving a space into the market, offering cheaper, standardised and safer inks.
• With this project, I want to harness the power and possibilities offered by Synbio to produce pre-existing bacterial pigments in a more efficient and scaled-up way. Although the initial idea for such pigments is for them to be used as the base for novel tattoo inks, their versatility and easy solubility could provide alternative dye uses down the road.
• By designing a genetic vector containing the necessary proteins required for pigment production and finding the optimal regulation, we could have pigment producing bacteria, with an easy purification process.

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A short overview

Strain development:

• Pigment selection and pathway research

• Vector selection, possible regulatory elements*

  *There is always the possibility of including elements for two different pathways sharing precursors, and using inducers to select for on/other

• Primer design, PCR and cloning strategy (Cell library of regulators)

• Growth assays, colorimetric assays for ideal strains

• Pigment extraction and purification

Health and safety assays for project:

• Fit with current regulation
• Pass any required biosafety tests
• Ensure safe production and proper training
• Efficient waste management
• Scale-up
• Test ideal culture conditions, ensuring production without toxicity
• Test bioreactors, turbidity conditions and aeriation

Marketability

• Public perception
• Ecological

Unexpected challenges

• Determine regulation for different constructs
• Determine lasting time
• Possible assumptions

Future Work

• The current objective is to develop the know-how to produce colorant compounds from biological sources and outsource the scale-up and purification processes. Once the purified product is obtained, the formulation could be handled in-house or further outsourced if necessary.
• Initially, the offered portfolio would be smaller and limited to specific existing bacterial compounds, with the hopes of eventually increasing, and using for example plant or fungal pigments. And through protein design, even produce custom orders for our clients. Maybe even obtain the capabilities for scale-up and purification in-house. </aside>

Background and motivation

• I am pursuing this project because it is an idea I first came up with 10 years ago, but never had the chance to properly develop, even after winning competitions with it and receiving praise for it.
• Currently, due to their multiple useful properties, bacterial pigments are being researched for their use in multiple different fields, such as for food safety, cosmetics, medical uses, or antibiotic properties. Many different pigments and their pathways are well-known, allowing for a good stating point for the project.

• There are other companies using biotechnology to produce color compounds, such as Colorifix, and huue, but they are focused on textiles. There is also LivingInk, focusing on the creation of algae-based pigments for different uses, none of which our own. To the best of my knowledge, this would be the one company to be involved in this particular type of production. And there is always the hope that our findings could be used in other settings as well, such as cosmetics or hair dyes.

SECTION 2: PROJECT AIMS

1. Outline three aims of your final project

1st Aim.

The first and more straightforward aim of my project is to obtain bacterial-produced pigments that could be properly purified and formulated in order to create an ink suitable for human tattooing. To do that, I would use different genetic constructs containing the sequences codifying for the proteins involved in the different pigment-producing pathways. In most cases, bacteria transformed with one construct would produce one pigment, but in cases where different pigments share precursor molecules, genetic switches could be incorporated to redirect the production in favour of the desired molecule.

These constructs can be designed in silico and commercially synthesised, then assembled via PCR and transformed. However, this approach is not as straightforward, as it would require a previous serious of tests to determine the ideal regulation mechanisms for each case. The most obvious way to do so is to establish a combinatorial library of regulators and genes, with the corresponding primers, amplify each compatible fragment, and automate the assembly process via Opentrons.

As a case study, one known genetic construct capable of synthesising the pigment violacein has already been ordered, and will be assembled, transformed, and tested at LifeFabs, as a way to confirm the required steps.