The Future of Food
May 25, 2020
Open-source 3D printing in COVID-19 response
May 27, 2020
Biotechnology Behind Culina
There is a need for a less resource-intensive practice
of meat, fish, and dairy production.
Feedstock production for agricultural farming accounts for 18 % of anthropogenic greenhouse gas emissions – a statistic coinciding with meat production, forecasted to more than double from 228 to 465 million tonnes as the population exceeds 9 billion by 2050 (1).
• The failure of plant-based start-ups to produce alternatives with a flavour and texture similtude to natural food products has discouraged consumers preventing prospective growth and replacement competition in the agri-food market.
• In line with current trends in biotechnology for animal-free protein production, a dynamic area of R&D means we can expect to see further optimisation of the technology used to achieve high yields, efficiency, and scale up.
• Yet, we differ to trends such as cellular agriculture as we still depend on microbes to express and secrete proteins which we harvest and develop into meat tissue with our perfusion bioreactor technology
• Pichia pastoris, a methylotrophic yeast, is an established eukaryotic chassis for metabolic engineering and protein expression on both laboratory and industrial scales (4).
• Factors making P. pastoris a competitive expression host include its secretory expression pathway reducing timely and costly means of protein separation and purification; stable genome integration of exogenous genes; and efficient downstream upscaling (4) - made possible due to advances in metabolic engineering and synthetic biology (5).
• Our 3D scaffold chamber with multilayering and spatial and temporal patterning allows harvested protein to reflect meat tissue topography. Petri dish grown meat is limited to ‘minced meat’ structures – we give our protein products a real meat feel.
• Opposed to commonly used gelatine scaffolds for tissue generation, we use synthetic or nature (chitosan/cellulose) biomaterials – we are completely meat-free.
• In vitro meat uses stem cell technology for clean meat production such as iPSC which require gene editing (10) – we want to offer a GM free product!
• In vitro meat maintains cell lines typically in animal sera such as fetal bovine serum (FBS) (11) – our yeast cell line is maintained by processing organic biomass from our hydroponic indoor form.
• In vitro methods face large-scale technical challenges such as cell source, culture media and yield (11). Yeast cell maintenance is well established, and secretion optimisation combats challenges with yield. Our products have no source to dead animals, thanks to the power of synthetic DNA.
• Our technology is absent of the ethical concerns surrounding embryonic, and adult stem cell technology used by most cell-based meat companies.
• In line with current trends in biotechnology for animal-free protein production, a dynamic area of R&D means we can expect to see further optimisation of the technology used to achieve high yields, efficiency, and scale up.
• Yet, we differ to trends such as cellular agriculture as we still depend on microbes to express and secrete proteins which we harvest and develop into meat tissue with our perfusion bioreactor technology
• Pichia pastoris, a methylotrophic yeast, is an established eukaryotic chassis for metabolic engineering and protein expression on both laboratory and industrial scales (4).
• Factors making P. pastoris a competitive expression host include its secretory expression pathway reducing timely and costly means of protein separation and purification; stable genome integration of exogenous genes; and efficient downstream upscaling (4) - made possible due to advances in metabolic engineering and synthetic biology (5).
• Our 3D scaffold chamber with multilayering and spatial and temporal patterning allows harvested protein to reflect meat tissue topography. Petri dish grown meat is limited to ‘minced meat’ structures – we give our protein products a real meat feel.
• Opposed to commonly used gelatine scaffolds for tissue generation, we use synthetic or nature (chitosan/cellulose) biomaterials – we are completely meat-free.
• In vitro meat uses stem cell technology for clean meat production such as iPSC which require gene editing (10) – we want to offer a GM free product!
• In vitro meat maintains cell lines typically in animal sera such as fetal bovine serum (FBS) (11) – our yeast cell line is maintained by processing organic biomass from our hydroponic indoor form.
• In vitro methods face large-scale technical challenges such as cell source, culture media and yield (11). Yeast cell maintenance is well established, and secretion optimisation combats challenges with yield. Our products have no source to dead animals, thanks to the power of synthetic DNA.
• Our technology is absent of the ethical concerns surrounding embryonic, and adult stem cell technology used by most cell-based meat companies.
We propose a biotechnology and biodesign approach for the development of microbial cell technology for sustainable meat production. The current ‘free from’ food market’s estimated worth is £837 million, according to Mintel (2018). However, failure to produce alternatives with a flavour and texture similar to natural food products has discouraged consumers, preventing prospective growth and replacement competition in the agri-food market. Therefore, current trends in R&D have embarked on metabolic engineering and synthetic biology solutions for meat and dairy production to tackle this climate-consumer crisis.
Meet the micromachinery behind recombinant protein production - yeast!
Meet the micromachinery behind recombinant protein production - yeast!
Meet the micromachinery behind recombinant protein production - yeast!
Historically, baker’s yeast, Saccharomyces cerevisiae, has been the epicentre of recombinant protein production in the biotech industry – valuing at tens of billions of US dollars each year (2).
Culina uses Pichia yeast cell factories to produce a high-value animal protein. A current target for protein production at Culina is crustacyanin (CRCN) protein (see below) found in Crustacea, Homarus gammarus, more commonly known as the European lobster, for shellfish meat alternatives such as crab, shrimp, and lobster (3). Protein design is fundamental to the process of meat production at Culina, see more here.
Culina uses Pichia yeast cell factories to produce a high-value animal protein. A current target for protein production at Culina is crustacyanin (CRCN) protein (see below) found in Crustacea, Homarus gammarus, more commonly known as the European lobster, for shellfish meat alternatives such as crab, shrimp, and lobster (3). Protein design is fundamental to the process of meat production at Culina, see more here.
Pichia pastoris, a methylotrophic yeast, is an established eukaryotic chassis for metabolic engineering and protein expression on both laboratory and industrial scales (4). Factors making P. pastoris a competitive expression host include its secretory expression pathway, reducing timely and costly means of protein separation and purification; stable genome integration of exogenous genes; and efficient downstream upscaling (4) - made possible due to advances in metabolic engineering and synthetic biology (5). The Pichia toolkit (PTK) (6), a development of the yeast toolkit (YTK) (7), consisting of genetic parts, will be used to assemble various recombinant Pichia expression systems and optimise design for protein titre and secretion.
Basic SBOL: Synthetic Biology Open Language of genetic parts based on the Pichia Synthetic Toolkit (6).
Pro-Yeast: Perfusion Bioreactor Technology
Our Pro-Yeast technology is in trend with tissue engineering bioreactors often used in combination with 3D scaffolds (7). Perfusion bioreactor technology has proven successful in tissue formation and quality (9), and in regard to up-scaling, similar technologies have developed cell volumes of up to 20,000 L (8), and scaffold technology has proven a success in cellular agriculture. Our perfusion bioreactor technology allows proteins secreted by our engineered yeast cell line to be harvested and sequestered through a perfusion protein pipeline. Proteins are transported into the surrounding culture medium and selectively filtered into the pipeline where they travel to a protein reservoir under high rates of perfusion. Proteins arrive at a second chamber, where larger target proteins are separated first by size-exclusion chromatography. With the help of gravity and additional vacuum enhancements, the protein is forced down the column, whilst smaller nutrient molecules are recycled back into the reactor vessel for continuous fermentation. Finally, protein molecules diffuse into a scaffold chamber where they bind to a macroporous 3D scaffold. The 3D matrix provides a structure alike that of meat tissue where immobilised proteins begin to collect - the product of 'clean' meat.
Our Pro-Yeast technology is in trend with tissue engineering bioreactors often used in combination with 3D scaffolds (7). Perfusion bioreactor technology has proven successful in tissue formation and quality (9), and in regard to up-scaling, similar technologies have developed cell volumes of up to 20,000 L (8), and scaffold technology has proven a success in cellular agriculture. Our perfusion bioreactor technology allows proteins secreted by our engineered yeast cell line to be harvested and sequestered through a perfusion protein pipeline. Proteins are transported into the surrounding culture medium and selectively filtered into the pipeline where they travel to a protein reservoir under high rates of perfusion. Proteins arrive at a second chamber, where larger target proteins are separated first by size-exclusion chromatography. With the help of gravity and additional vacuum enhancements, the protein is forced down the column, whilst smaller nutrient molecules are recycled back into the reactor vessel for continuous fermentation. Finally, protein molecules diffuse into a scaffold chamber where they bind to a macroporous 3D scaffold. The 3D matrix provides a structure alike that of meat tissue where immobilised proteins begin to collect - the product of 'clean' meat.
3D Scaffold Fabrication: Solvent Casting Method
- Solvent casting fabricates a macraporous scaffold; other methods include 3D printing and electrospinning (7)
- Polymer matrix creates structural architecture and network of support for macromolecule binding (7)
- Multilayering and spatial and temporal patterning reflects tissue topography (7)
- Choice of biomaterial: synthetic polymers, natural biopolymers e.g. cellulose or hybrids of the two (7)
- Synthetic polycaprolactone (PCL) is FDA approved and will allow mechanical dissociation of the final 3D protein product (10)
- Polymer matrix creates structural architecture and network of support for macromolecule binding (7)
- Multilayering and spatial and temporal patterning reflects tissue topography (7)
- Choice of biomaterial: synthetic polymers, natural biopolymers e.g. cellulose or hybrids of the two (7)
- Synthetic polycaprolactone (PCL) is FDA approved and will allow mechanical dissociation of the final 3D protein product (10)
All that is now required of you is to cook in your choice of specialist rooms!
References:
1. Z. B. Bhat et al., Journal of Food Science and Technology 48, 125-140 (2011).
2. M. Huang et al., PNAS 115, 11025-11032 (2018).
3. N. M. Wade et al., Molecular Biology and Evolution 26, 1851-1864 (2009).
4. T. Zhu et al., Biotechnology Journal 14, 1800694 (2019).
5. O. P. Ishchuk et al., Recombinant Protein Production in Yeast 1923, (2019).
6. U. Obst et al., ACS Synthetic Biology 6, 1016-1025 (2017).
7. S. Ahmed et al., Biotechnology Letters 41, 1-25 (2019).
8. R. P. Harrison et al., Biointerphases 12, (2018).
9. J. Zhao et al., Biochemical Engineering Journal 109, 268-281 (2016).
10. T. B. Arye et al., Front. Sustain. Food Syst (2019).
11. S. J. Allan et al., Front. Sustain. Food Syst (2019).
1. Z. B. Bhat et al., Journal of Food Science and Technology 48, 125-140 (2011).
2. M. Huang et al., PNAS 115, 11025-11032 (2018).
3. N. M. Wade et al., Molecular Biology and Evolution 26, 1851-1864 (2009).
4. T. Zhu et al., Biotechnology Journal 14, 1800694 (2019).
5. O. P. Ishchuk et al., Recombinant Protein Production in Yeast 1923, (2019).
6. U. Obst et al., ACS Synthetic Biology 6, 1016-1025 (2017).
7. S. Ahmed et al., Biotechnology Letters 41, 1-25 (2019).
8. R. P. Harrison et al., Biointerphases 12, (2018).
9. J. Zhao et al., Biochemical Engineering Journal 109, 268-281 (2016).
10. T. B. Arye et al., Front. Sustain. Food Syst (2019).
11. S. J. Allan et al., Front. Sustain. Food Syst (2019).
