To demonstrate the power and versatility of the precision fermentation approach, we used microbial fermentation to produce three model proteins of potential value for human nutrition.
These protein examples come from a plant, an animal and from a probiotic gut bacterium respectively. As every microbial platform has its preferences when it comes to expressing proteins, we took advantage of our own range of production platforms to choose the ones best suited.
Case 1
Developing a plant protein with great taste
Goal: to provide a low-calorie, plant-based, high potency sweetener as a sugar substitute
Origin: edible plant
Size: 6.5 kDa
Host: Pichia pastoris
Status: developmental
Partner: open
Case 2
Developing a microbial protein for nutrition
Goal: to turn a microbial protein from fermented food or healthy human gastrointestinal flora into a source of nutrition in its own right
Origin: human gut bacterium
Size: 70 kDa
Host: Aspergillus niger
Status: developmental
Partner: open
Case 3
Developing an animal protein sourced from a microorganism
Goal: to replace an animal protein of desirable functionality and/or nutritional value with an identical mimic produced by microbial fermentation
Origin: farm animal
Size: 15 kDa
Host: Yarrowia lipolytica
Status: commercial
Partner: portfolio Biocatalysts
Our approach:
Having identified a desirable protein of choice that matches a list of criteria including functionality, safety and nutritional value, the next key step is selecting a matching microbial production host from our range of platforms.
Our production hosts are the result of continuous improvement programs at BRAIN involving a genetic toolbox, proprietary genome editing and a wealth of experience.
Promising initial productivities seen for a target protein are further improved by developing suitable fermentation and downstream processes. These need to match requirements for cost-efficient production when scaled up to commercial scale at our ISO-certified production plant or a selected toll manufacturer.
Every microbial expression platform has its performance characteristics, strengths and weaknesses. It is therefore critical to select the right platform that matches specific requirements for a given target protein.
Production strains are selected from bacteria (E. coli, Bacillus), yeasts (Pichia, Yarrowia) and filamentous fungi (Aspergillus). Depending on provenance and category of the target protein in question, an initial host screening will be performed to determine preferable choices from BRAIN’s range of platforms.
Strain
development
Strain development
The development of a microbial production host targets its biosynthetic machinery, which is critical in defining its performance as an efficient protein producer. Complex cellular processes governing gene expression, protein folding, secretion and stability involve an array of factors that may need to be introduced or improved. Our experience in setting up expression systems, a comprehensive genetics toolbox and our proprietary genome editing system are key to achieving these goals.
Process
development
Process development
Gene expression and protein secretion are natural processes of microbial physiology determined by the cell’s genetic setup, available nutrients and environmental stimuli. Critical parameters for a fermentative production process therefore include growth media composition, feeding rates, agitation and aeration, all of which require careful monitoring and adjustment to ensure process productivity.
Production
Production
Scaling up a microbial fermentation process for protein production from lab scale to commercial production can literally mean moving from drop volumes to swimming pool dimensions of controlled fermentation media, with living cells that need to be kept happy and productive.
Within the BRAIN Biotech Group we are equipped to perform submerged fermentation up to the 10 cbm scale using ISO-certified equipment and processes. We have established partnerships for going larger.
The results
A great-tasting protein
Brazzein is a small, sweet-tasting plant protein and is 1000x sweeter than sucrose. The high-potency sweetener brazzein illustrates the potential of alternative proteins produced by precision fermentation to be used as a valuable food ingredient to improve taste. This is relevant because sucrose, which is commonly used, is known to have negative health effects when consumed in excess.
A protein for precise nutrition
Protein A1 is an example for taking a microbial protein and turning it into a source of nutrition. Naturally produced by a health-promoting (probiotic) microbe of the human digestive system, A1 is a candidate for evaluation as a food ingredient.
A protein to improve food functionality
Originally sourced from animals, this functional protein was precisely produced by microbial fermentation in yeasts and is used to improve food functionality by modifying fats in egg yolk. Acting as an enzyme, this particular protein’s function is limited to the food preparation process (processing aid) and is inactivated before consumption.
Are you looking for support in making #FutureFood a reality?
Tap into the competencies and resources of the BRAIN Group! Don’t hesitate to reach out to us. Our expert for alternative proteins is Patrick Lorenz.
More about our capabilities for alternative proteins
Enabling #Futurefood
Nutrition in industrialized countries will move away from meat and dairy consumption towards alternative sources of protein and foods. These will rely on plants, microorganisms and cell cultures. Biotechnology will be the key enabler. This is where we come in.
