Manufacturing Deep Dive: New frontiers

A process thousands of years in the making is receiving a high-tech twist, allowing food producers to create ingredients that address concerns such as sustainability, nutrition and animal welfare. David W. Smith catches up with 10 innovators who are pushing the boundaries of this ancient practice

Over-reliance on animals to provide enough protein to feed the world will become unsustainable as the population rises to 10 billion around 2050. There simply isn’t enough agricultural land to make it viable. But fermentation – and in particular precision fermentation – offers a solution. The technology uses microbial hosts as ‘cell factories’ to produce specific functional ingredients and is essentially a refined form of brewing used to create high-protein foods with low environmental impact. There are a host of innovative start-ups working independently or sharing their protein powders with food giants such as Mars, Nestlé and Starbucks.

Precision fermentation produces an almost limitless range of foods. When the microbes are fed on hydrogen, or methanol, they can create a flour with at least 60% protein content, which is three times the concentration found in chickpeas. They can also be bred, or programmed genetically, to produce replacements for fish, milk and eggs with superior taste, texture and nutrition to the plant-derived vegan options.

Environmental claims

The environmental footprint of precision fermentation is drastically lower than for conventional agriculture. Researchers from the Swedish University of Agricultural Sciences, for instance, estimated it used 1,700 times less land than soy grown in the USA. The gulf with farming for beef and lamb is even greater – more than 100,000 times less land is required for precision fermentation. Meanwhile, the reductions in water and GHGs are huge, especially if renewable sources of energy are used.

There is so much innovation happening in this corner of food-tech, especially in precision fermentation. Experts have created casein proteins that accurately recreate the taste of cow’s milk, or the flavor and stretchiness of cheese. Mycelium is being used as a source of high-protein powders to make meat substitutes rather than using wheat, peas or soy.

The sector has attracted eminent scientists from the fields of medicine and engineering and the creativity is ready to burst out. Some constraints remain, however. There are regulatory issues, although barriers are falling. More serious is the issue of scaling: the lack of precision fermentation technology keeps prices high.

As the issue of scaling is resolved and regulatory barriers fall, though, the potential will be felt. Imagine if you will a microbial brewery in every town making cheap protein-rich foods for local people. This would be a game-changer when it comes to the environmental issues around shipping food thousands of miles, as well as help geographical areas where land is barren, and water is scarce.

JARED RAYNES, HEAD OF PRECISION FERMENTATION & DAIRY, ALL G FOODS

Jared Raynes is reputedly the first scientist in the world to recreate the protein structure of milk using DNA sequencing from cows yet without the requirement of an animal. He achieved the feat just over a decade ago using precision fermentation. Nine years later, in 2021, he was hired as Head of Precision Fermentation & Dairy at All G Foods in Australia to transfer his foundational casein and dairy science research into animal-free dairy products. Then, in 2022, he was promoted to become the company’s Chief Scientific Officer.

Raynes was born in New Zealand and studied Biochemistry & Microbiology at the University of Canterbury in New Zealand under the renowned Professor Dame Juliet Gerrard. But it was after he relocated to Australia in 2012 and joined the Food Program of the country’s Commonwealth Scientific and Industrial Research Organisation (CSIRO) that he began to work on the structure and function of casein proteins, the major protein component of milk.

“I began generating casein proteins using precision fermentation in 2013, which was before the establishment of the first dairy precision fermentation company in 2014,” Raynes states. “With precision fermentation, I could precisely control the experimental design to see how the casein proteins assemble into the casein micelle, the large protein structure that binds calcium and phosphate and gives dairy its amazing functionality.”

Raynes was hired by the Czech Republic-born entrepreneur, Jan Pacas, who was searching for the best scientists in the field after founding All G Foods in 2020. Pacas had previously been CEO of a Swiss multinational but after relocating to Australia, he founded two successful companies, Mad Paws (Asia’s largest online marketplace for pet services) and Flare (one of Australia’s fastest-growing fintechs).

We believe we stand out from competitors due to our expertise in metabolic engineering, dairy science, protein engineering, fermentation and, in particular, the production of phosphorylated casein proteins using precision fermentation

Credit where it’s due

The success of those businesses gave Pacas credibility with the financiers and All G Foods secured backing from Clean Energy Finance Corporation (CEFC), Ellerston Capital, and Triple Star Capital, among others. In a short time, All G established its plant-based meat brand, BUDS!, in burger, mince and sausage versions. But Pacas had more radical ambitions: he wanted to develop animal-free bioidentical dairy products using precision fermentation.

“I have now been working with precision fermented caseins for 10 years, which we believe is one of the longest in the field, and we have Dr Carl Holt as a sole advisor to All G Foods,” Raynes explains. “He was affiliated to Glasgow University and is recognized as the number one casein researcher in the world. We believe we stand out from competitors due to our expertise in metabolic engineering, dairy science, protein engineering, fermentation and, in particular, the production of phosphorylated casein proteins using precision fermentation.”

This nascent industry faces several stumbling blocks, Raynes notes, but all should be resolvable. The issue of scalability is the main bugbear and there are likely to be bottlenecks for some companies for years to come. This is especially true in new markets where precision fermentation technology may not yet exist. “Through collaborations, the establishment of specialized training programs to build a competent workforce, and both public and private investments in large-scale facilities, these obstacles can be overcome,” he says. Meanwhile, All G is fortunate that one of its chief investors, Agronomics, has a company in its portfolio called Liberation Labs that has started building dedicated precision fermentation facilities.

There are also hurdles related to approvals but Raynes says it is a question of educating governments on the safety and quality of this new food technology. “Our regulator, Food Standards Australia New Zealand, is currently reviewing the definitions for ‘food produced using gene technology’,” he reveals.

Raynes expects rising demand for the technology as global sales of dairy increase. He is also encouraged by both the positive public reaction to the company’s fermentation strategy and investor confidence. “We get a tremendous response at public events, and we have strong investors who have been on board since the beginning because they share our vision for the future,” he says. “We are single-minded in our focus to become the Asia-Pacific region’s precision fermentation leader.”

ADAM LEMAN, LEAD SCIENTIST, THE GOOD FOOD INSTITUTE

“Right now, 77% of arable land is used for animal agriculture, which gives us about 23% of our protein supply. These numbers are poor and won’t work when the population increases by 25%,” says the Good Food Institute’s Adam Leman, Lead Scientist.  ”Our planetary resources will not allow us to continue on this trajectory for the next 27 years and beyond. This is where microbes can make a difference.”

An entire cow weighing hundreds of kilograms must be raised and fed before it can calve and produce milk, Leman adds, whereas microbes can be rapidly grown in a tank and induced to produce a protein or fat ingredient. “Microbes are also incredible at biosynthesis,” he says. “With just a few essential ingredients such as sugars, nitrogen and minerals, they can produce complex molecules like proteins, fats and pigments. If we move to an efficient system where microbes make these specialized ingredients without the massive inefficiencies of animal agriculture, then the utility to the food system for each precision fermentation nutrient will increase.”

Achieving scale

The biggest challenge, Leman points out, is scaling. Fermentation-derived fuel produces enormous amounts of ethanol and pharmaceuticals derived from the technology produce life-saving medicines like insulin. But until a few years ago, little attention was paid to food precision fermentation except for some food-preparation and processing enzymes. “There is now an opportunity to scale out at the level of industry and to open fermentation facilities worldwide as local production hubs. We could build facilities in the hundreds of thousands to millions of liters to enable economies of scale.”

Right now, 77% of arable land is used for animal agriculture, which gives us about 23% of our protein supply

But more R&D is required to optimize the microbes, their nutrients, the fermentation vessel design, and downstream processing equipment. “All this is going to take some time and capital, so it’s going to be dependent on finding public and private investment to build the infrastructure and the biotechnology,” Leman says. “Food needs to be made at a scale close to the fuel fermentation industry, but at the same time it needs to be safe and pure in a manner closer to pharmaceutical fermentation.”

Although precision fermentation is in its infancy as a food producer, the sector has brought in a lot of talent on the biology and engineering sides who are drawing on historical knowledge from the fermentation industry as well as food science. “Once we can harness that talent and build out the industry, there will be job creation for facility operators, new opportunities for growers to bring in their feed-stocks, and creative ways to continue to innovate,” Leman suspects.

The trend is for precision fermentation to replace common ingredients such as dairy or egg proteins, fats and oils. “This makes a lot of sense as these ingredients can be ‘dropped in’ to existing products,” the GFI man notes. “On the dairy side, whey proteins such as beta-lactoglobulin are replacing animal-derived proteins in ice cream and spreadable cream cheese. Meanwhile, ovalbumin is the primary protein in egg whites and is being produced by precision fermentation.”

The industry is also moving closer to bringing dairy proteins such as lactoferrin and casein proteins to market. Lactoferrin is a small fraction of the dairy protein profile but research is building evidence of its ability to defend against infections. TurtleTree has announced it will bring a bovine lactoferrin protein made by precision fermentation to market in 2023.

Precision fermentation also has a role in the wider bioeconomy, such as recycling food waste. Right now, the industry is mostly using highly pure sugars as a carbon source. But while microbes are proficient at biosynthesis, they’re also adept at breaking down things like peels, stalks, hulls, and lots of carbon-rich plant matter that we currently consider food waste.

“Microbes can break down those starches into usable sugars and take smaller carbon-containing molecules like ethanol or acetate and convert them into energy,” Leman advises. “If we use some of the precision fermentation microbes or even upstream microbes to convert these other carbon sources, sustainability increases substantially. Another area the industry is actively working on is to make use of the biomass – the cells left over after fermentation. These cells contain lots of nutrients that could feed a separate fermentation for a bioplastic or citric acid, or new processing techniques could allow us to use them as fertilizers.”

STAFFAN HILLBERG, CHAIRMAN, MILLOW

Swedish start-up, Millow, has developed a fermentation system to create a nutritious hybrid protein out of plants and mycelium in fewer than 24 hours. The approach is based on more than 25 years of research into mycelium microorganisms by Mohammad Taherzadeh, Professor of Biotechnology at the University of Borås in Sweden, who designed a system of ‘Controlled Environment Fermentation’. This consists of custom hardware known as the ‘S-unit’ and accompanying software.

The substrate and the mycelium is inserted into the S-unit and after 12-24 hours, it produces a finished product. Depending on the ‘recipe’, it might require minimal post-processing.

“As we can control every detail of the environment within the S-unit when it comes to temperature, humidity, time, gases, liquids, we like to call this ‘Controlled Environment Fermentation’ in a similar way to ‘Controlled Environment Agriculture’ for growing vegetables,” reveals Staffan Hillberg, Chairman.

The software can be programmed to subtly adjust the process to create different products. “We can influence aspects such as texture, taste, how the mycelium grows and in what direction, which parts of the substrate it uses and how,” Hillberg adds. “We’re also using artificial intelligence and self-learning algorithms to continuously improve production. The details are a trade secret.” Meanwhile, compared to traditional mycelium production techniques, the S-unit requires a third of the energy, a fraction (2.3%) of the water, and emits 95% less CO2.

Perfecting texture

A great difficulty in food-tech is creating the right texture, Hillberg points out. The artificial meat sector’s struggles with the issue have led many companies to create ultra-processed foods packed with additives and binders. Millow has developed a bioengineering method known as SHORT (Mycelium Utilised Texture Engineering) that uses mycelium filaments to engineer the texture. “The appealing texture allows it to be used in most types of food handling and it can even be boiled in stews without dissolving or losing its structure,” he says.

We’re also using artificial intelligence and self-learning algorithms to continuously improve production. The details are a trade secret

“It’s also important to us that it’s completely natural,” Hillberg continues. “We don’t understand why anyone would produce an alternative protein where humans can’t advantage of minerals, vitamins and other elements. They need to be at least as healthy as traditional food, or there’s going to be a backlash.”

Millow has been in ‘stealth mode’ for almost two years. During this time, the team has met with several food companies in the Nordics and prepared more than 10 different products. Recently, an agreement was signed with a large producer to bring two foods to market.

“Although the strain of mycelium we’re using does not require novel food approval, we have access to hundreds of strains that change parameters like texture, taste, color and nutrition,” Hillberg reports. “We’re focusing on oats but we’ve done trials with many other base substrates – even some food waste streams. We see an opportunity for more circular processes that lower costs.”

The initial target for Millow is food producers with a B2B strategy but with investment, it could develop a B2C strategy. An early collaborator and shareholder is Rob Janoff, famous for designing the Apple logo. His team created Millow’s name and brand identity. ”Despite the B2B strategy, it is seen as more than a simple ingredient by food producers and will be co-branded.

“There is a great deal of research and experimentation into fermentation, 3D-printed food, and cultivated meat, attracting capital and creating a positive environment for growth. We expect alternative proteins to become at least as large as traditional proteins.”

And by 2030, Hillberg predicts Millow to be well-established in the Nordics with several products and to have entered other markets. “There’s a possibility we will have spun out a separate business focusing on fish feed and pet food.”

TIM GEISTLINGER, CHIEF SCIENTIFIC OFFICER, PERFECT DAY

The growing profile of the precision fermentation sector is signaled by its ability to attract high-flying scientists from the world of medicine. California-based Perfect Day – whose animal-free milk is being used by firms such as Nestlé and Mars – hired its Chief Scientific Officer, Tim Geistlinger, after he’d made major breakthroughs in treatments for HIV-AIDS and cancer.

Geistlinger left that distinguished career in medicine in 2013 to become the first scientist at plant-based food company, Beyond Meat, where he built the R&D team from scratch. “I loved the vision of creating healthy alternatives to animal meat but also to help save our planet from the negative impacts of the meat industry,” he says. ”Launching the Beyond Burger range was “much more fun than medicine”, he notes.

A meeting of great minds

In 2016, a new challenge arose after he met Ryan Pandya and Perumal Gandhi, the co-founders of Perfect Day. The two men – bioengineers and vegans – had begun to explore precision fermentation as they were frustrated at the “lacklustre experience” of dairy alternatives. A few years earlier, they had founded Muufri, which used genetically engineered yeast to produce lactose-free milk. Having identified the DNA blueprint in literature from the dairy industry, it required no animal involvement.

Pandya and Gandhi changed Muufri’s name in 2016 after a study showed cows produced more milk when listening to the Lou Reed song Perfect Day. It was at that stage that Geistlinger came on board. “I was excited by their vision,” he recalls. “Together, we took an idea, built a platform, scaled up, and manufactured our first products. Multiple companies are now using it and our products are in 50,000 stores.”

Nestlé developed its animal-free ‘Cowabunga’ milk in Switzerland with its R&D team using Perfect Day’s animal-free whey. The ‘milk’ landed in select Safeway stores in San Francisco back in December 2022 in original and chocolate flavors. It was Perfect Day’s fourth collaboration on fluid milk products. It has previously launched with Betterland Foods, Tomorrow Farms, and Strive Nutrition.

We’re scaling by joining forces with food partners, including Mars and Bel Group, and food-service partners in campuses across the USA

Perfect Day has also worked with General Mills on its Bold Cultr cheese, Mars on milk chocolate bars, and Starbucks to test fluid milk in select Pacific Northwest stores last year.

Geistlinger expects further growth and adoption of Perfect Day’s products and services across its three business units in 2023. The first unit is B2B ingredient innovation with new partners using the dairy protein. The second is new consumer products through subsidiary, The Urgent Company. Third is the enterprise biology business, nth Bio, which allows companies to leverage their technology and understanding.

One of the greatest challenges for Perfect Day, Geistlinger concedes, is communicating to food producers the science behind their twist on making dairy protein. “Once we explain how we’re using fermentation to convert sugar into high-value, pure, functional, true-to-nature proteins – and how this will allow them to not only make their favorite products without any sacrifice but also meet their environmental goals – they’re very interested,” he says. “When they try the products and realize that they do not need to change operations, they love it.”

The issue of finding manufacturing infrastructure to support the industry’s growth is also being addressed. “Last year, we scaled our production three times, but we are also actively exploring building our owned manufacturing capabilities to allow us to produce tens of thousands of metric tons in the next five years,” he says. “We’re scaling by joining forces with food partners, including Mars and Bel Group, and food-service partners in campuses across the USA. We’re growing in places such as India – a critical market for us to scale our production capacity. In fact, the Food Safety and Standards Authority of India has already approved our application for milk protein from fermentation, opening the door for commercialization there.”

MORAN FARHI, VP OF RESEARCH, BETTER MEAT CO

The Sacramento-based Better Meat Co has created what it deems a fourth major ingredient that can act as a meat substitute. Most manufacturers rely on wheat, peas and soy but Better Meat has patented a method using fermentation to produce its ‘Rhiza’ mycoprotein from mycelium.

“Rhiza can easily be used as the basis for animal-free meat and blending with animal meat at lower-than-animal meat cost,” suggests Moran Farhi, VP of Research. “We pursue a B2B model which means we’re not trying to disrupt the meat industry but work with it.”

Feedback from partners, including Hormel Foods, Perdue Farms, and Maple Leaf Foods, led the company to develop a white ingredient that is neutral in flavor and packed with nutritious protein and fiber. “This functions as a blank canvas that food scientists can turn into different applications such as beef, chicken or fish alternatives,” Farhi reports. “Making a dry product that retains a white color is not trivial and we have recently identified and modified key aspects in our process to routinely produce large amounts of pristine product.”

Rhiza is made from a fungus species known as Neurospora crassa. In Indonesia, it’s used to ferment pressed soy, peanuts or coconuts into oncom, a tempeh-like staple food. But it’s not previously been cultivated to be eaten on its own. Farhi says it has many advantages, not least that it is meat-like in texture and grows rapidly. From a nutritional perspective, it contains more protein than eggs, more iron than beef, more fiber than oats, more potassium than bananas, and is a natural source of B12. Its flexibility means it has already been turned into substitutes for bacon, chicken nuggets, lunch meat, foie gras, and even used as a dairy and egg substitute in baking applications.

The genetic improvements we seek are also unconventional as we maximize abstract attributes such as texture, flavor and aroma

A tested development

Developing the processes to produce Rhiza was far from straightforward. Most fermentation is by-product of cell growth, whereas Better Meat needed to produce biomass from fungi to imitate the texture of meat. “We needed to develop a process that puts an emphasis on maximizing the total protein production and not a defined target, and we don’t want it to be secreted to the media,” Farhi says. “The genetic improvements we seek are also unconventional as we maximize abstract attributes such as texture, flavor and aroma. Our fermentation process also needs to be optimized to maintain color, texture, and nutritional values within the cells and not a function of one molecule.”

Operating in such a new field has both advantages and disadvantages for scientists and engineers. “The disadvantage is there’s not a lot of know-how, literature, expertise, or tools,” Farhi says. “But we’re building the knowledge and technology needed within the team. The advantage we have is we have a lot of room to innovate!”

Farhi believes the industry would benefit greatly from a change of mindset in the education system to prepare PhD and post-doc researchers to work in manufacturing. “Even industrial R&D departments often work siloed from manufacturing and don’t have direct communication with engineers,” he says. “That means that a lot of bioprocess R&D is developed in a way that does not translate at scale. Closing the gap between labs and production – between scientists and engineers – could lead to a mindset change in which R&D considers any scale-up limitations at the get-go.”

Better Meat’s 13,000ft2 facility in California can produce thousands of pounds of Rhiza a month, and the company is also expanding its R&D. “There are quite a few regulatory hurdles as it’s a new field, but I’m optimistic about the future based on the rapid advancements we’ve made, the financial investments in the company, and the positive feedback from customers.”

PATRICK LORENZ, VICE PRESIDENT OF STRATEGIC INITIATIVES BIOSCIENCE, BRAIN BIOTECH

Last year, the German industrial biotechnology specialist, BRAIN Biotech, teamed up with Formo to develop animal-free milk proteins. Formo is using the proprietary BRAIN Engineered Cas (BEC) gene-editing tool to optimize its microorganisms for commercial consumption using precision fermentation. “We deal with the entire value chain for protein production from candidate discovery to expression to fermentation scale up to manufacture,” explains Patrick Lorenz, BRAIN Biotech’s Vice President of Strategic Initiatives Bioscience. “Serving B2B industries, we also support customers in navigating their product’s regulatory journey. This is critical in this sector and particularly for precision fermentation.”

BRAIN Group started as a service provider for biotechnology in the 1990s, discovering and expressing enzymes and developing cell-based assay systems for the detection of small bioactive molecules responsible for taste. Both activities grew organically through mergers and acquisitions.

“With our knowledge of enzyme discovery, engineering and expression we can optimize and modify protein properties,” reveals Lorenz. “As not every protein can be expressed equally well in a given host system, our choice of appropriate microbial platforms is critical. We can then develop a protein production process including downstream processing up to the 60m3 scale,” Lorenz says. “These facilities are continuously expanding. We also recently launched proprietary gene-editing CrispR technologies to facilitate production strain development.”

Lorenz believes precision fermentation has the potential to provide any of the many types of protein that microbial systems are capable of synthesizing. The sustainability benefits are clear, too, he feels. “The environmental footprint of fermentation-derived proteins as obtained by precision fermentation, particularly if using renewable energy, is in an entirely different ballpark compared to current conventionally sourced proteins – both meat and dairy,” he adds. “On top of this come added direct and indirect health benefits, such as avoiding unhealthy ingredients, zoonotic diseases, and the spread of antibiotic resistance.”

With our knowledge of enzyme discovery, engineering and expression we can optimize and modify protein properties

But there are barriers to overcome to achieve commercial success on a more meaningful scale. “The critical thing is for alternative proteins to deliver on the three Ps – parity of taste, parity of texture and parity of price – with conventionally produced foods and food ingredients,” Lorenz continues. “If precision fermentation proteins are to replace their animal-derived siblings, it is likely they will be identical in performance. But as soon as price parity is discussed, I view the possibility with caution. When precision fermentation advocates propose a general trajectory leading to prices of very low double-digits in US dollars per kg of protein, I believe that will be the exception for only a few outstanding proteins.”

Building strains and acceptance

BRAIN has built a large collection of microbial strains by identifying microbes capable of fermenting edible food side streams. They can be used to produce condiments, ingredients and probiotic microbes for gut health. Lorenz argues that if precision fermentation struggles to deliver desirable proteins at reasonably economic costs, other strategies might be considered. “I’m thinking of unconventional food-associated proteins, or complementary strategies such as using whole microbial cells,” he says.

Widespread acceptance of precision fermentation among the public is another critical matter for the industry. For this reason, some of the biggest players in the field have teamed up to form the Precision Fermentation Alliance, whose goal is to promote the industry. BRAIN Bio’s partners in the Alliance include Remilk, Perfect Day, Change Foods, Imagindairy, New Culture and The Every Company.

It’s not difficult to imagine a positive future for the sector, but the precise trajectory it will follow is harder to predict. Lorenz expects a lot of experimentation with different technologies in the alternative proteins sector that could affect the direction it takes. “It’s conceivable we could get technology hybrids that – assuming they are accepted by the regulators and the public – would be game changers, including things like GM-improved single-cell proteins containing nutritious proteins.”

IRINA GERRY, CMO, CHANGE FOODS

Food technology start-up, Change Foods, thinks the biggest difference it can make to the dairy sector is in cheese. The ambition of the precision fermentation company – which is headquartered in both California, USA and Australia – is to create animal-free pizza cheese that tastes as good as the animal-derived versions.

Not only is the cheese sector the third-largest contributor of GHG emissions from agriculture sources, but even committed vegans find that plant-derived cheeses – usually made from soy, nuts, or peas – are unsatisfactory in terms of flavor and texture. Change Foods expects to launch its first products onto the market in 2024.

“Cheese is where consumers feel the biggest gap between their desire for a more sustainable food system, and the quality of the plant-based options available today,” explains Irina Gerry, CMO. “By producing milk proteins such as whey and casein through precision fermentation, we can offer products with comparable taste, texture and nutrition, but with a much smaller footprint.”

Gerry says that quite a few precision fermentation companies are focusing on whey production, but only a handful have cracked the more difficult code required to make casein – the dairy protein that gives cheese its flavor and texture. “By making casein through precision technology, we can create animal-free mozzarella cheese that melts and stretches on a pizza just like the cow-derived versions,” she says.

Cheese from cows is right at the top of the list when it comes to water-intensive food production. Academics Joseph Poore and Thomas Nemecek published research in the prestigious Science journal that revealed it took 5,605 liters of water to produce a kilo of cheese. This was more than twice as much water as the 2,714 liters required to produce a kilogram of beef, the most water-intensive meat product. “Unfortunately, global milk production continues to rise steadily as more consumers ascend into the middle-class and adopt western-style diets,” Gerry comments.

Complex picture

In the USA, the picture is a bit more complicated. Liquid milk consumption has been declining, but total dairy consumption remains steady according to the USDA. Gerry says this is because sales of cheese have risen. “It takes about 10 liters of milk to make just 1kg of cheese, so it’s easy to see how a decline in liquid milk sales is offset by a small increase in cheese sales.

By producing milk proteins such as whey and casein through precision fermentation, we can offer products with comparable taste, texture and nutrition, but with a much smaller footprint

We cannot solve the environmental issues associated with global milk production if we do not address cheese consumption,” she argues. By contrast, Change Foods’ technology uses 100 times less water, 10 times less land, and five times less energy than the animal-based version of cheese.

Change Foods was founded in 2020 and has raised more than US$15 million to date. Towards the end of 2022, it announced a deal with Kezad Group to build a 1.2-million-liter fermentation facility in the UAE. The company estimates it will replace the output of 10,000 dairy cows. The location in Abu Dhabi was chosen strategically to provide easy shipping and distribution to both Middle East and Asia-Pacific partners.

Gerry cautions this nascent industry must be careful to keep the end consumer in mind. Using precision fermentation to produce key functional ingredients, such as proteins and fats, has many benefits for the environment, animal ethics and nutrition, but comes with risks. “The main dangers are consumers who are allergic to dairy if the marketing doesn’t make clear exactly how precision fermentation works,” she warns. “As milk proteins made via fermentation are bio-identical to those from a cow, someone with an allergy to whey or casein is likely to have the same allergy when they’re made using precision fermentation. Simple and clear communication is a must.”

The need for clear and consistent messaging inspired, in part, the creation of the Precision Fermentation Alliance. Launched in February by nine leading precision fermentation tech startups, including Change Foods, the alliance will work to promote understanding of precision fermentation and educate and engage key stakeholders throughout the food industry value chain to establish best practices regarding regulatory, manufacturing, food safety, and communications standards and compliance.

AVIV WOLFF, CO-FOUNDER, REMILK

Childhood connections inspired the Israeli entrepreneur, Aviv Wolff, to co-found the animal-free dairy company, Remilk, in 2019. Wolff listened as a boy to the stories of his grandfather who was a dairy farmer. “I grew up understanding both the desire for dairy, and the heavy costs associated with it both on animal welfare and sustainability,” he says. “Years later, I wanted to take part in an initiative that could transform the way that we make our food and feed our planet’s growing population without destroying it in the process.”

In 2019, Wolff met Dr Ori Cohavi, a biochemist specializing in protein research, and they founded Remilk. They both agreed that most people were unwilling to give up foods they have always enjoyed, but that the traditional methods of producing dairy were terrible for animals and the planet. They set out to create a more sustainable product with the same quality, taste and texture as dairy milk. “As people don’t want to give up the taste and texture of dairy, we decided on a simple approach. We change the process not the food.”

The pair adapted the age-old fermentation method to recreate ‘cowless’ dairy proteins that are identical to their traditional counterparts. “Our new twist was to copy the genetic sequence – or ‘milk-making recipe’ – from a cow, inserting it into yeast and transforming these tiny, one-celled yeasts into a milk protein-making factory,” says Wolff. “The result is identical, nutritious milk proteins that form dairy products with the same taste and texture, but without lactose, cholesterol, hormones and antibiotics.”

It takes up to 1,000 liters of water to produce just one liter of milk, whereas our process requires around 1% of the land and 4% of feed, and less than 5% of the water

Replacing a 600kg animal with single-celled yeast to produce milk will make a radical difference to the sustainability of the planet, Wolff argues. “Animal agriculture relies heavily on the earth’s limited resources and is an important contributor of greenhouse gas emissions as well as the leading cause of deforestation in the Amazon rainforest. “It takes up to 1,000 liters of water to produce just one liter of milk, whereas our process requires around 1% of the land and 4% of feed, and less than 5% of the water. It emits a fraction of the greenhouse gases and it’s extremely scalable, enabling us to shift quickly from lab-scale to commercial-scale production. It’s also possible to make it anywhere in the world, whereas staple foods such as dairy are hard to produce in large quantities in a lot of geographies where heat conditions compromise the ability of cows to produce milk.”

Financially backed

Since 2019, Remilk has attracted huge investment, raising US$130 million in two fundraising rounds. In January, it announced a partnership with the US food giant General Mills – which owns brands such as Cheerios, Yoplait, Pillsbury and Haagen-Dazs. General Mills’ innovation unit has begun using Remilk’s animal-free whey protein and is already marketing a vegan cream cheese. It is Remilk’s first commercial product launch and a debut in the US market.

More good news came in February this year when Remilk received regulatory approval for sales of its protein from the Singapore Food Authority. At the same time, it received a ‘No Questions Letter’ from the US Food and Drug Administration. It indicates the unanimous opinion of an expert panel that Remilk’s whey protein can be safely used in food products under its GRAS (Generally Recognised as Safe) standards. Wolff believes the approvals were confirmation that Remilk’s animal-free milk was bioidentical to cow-derived dairy and safe for consumption.

With regulatory hurdles clearing, there is potential for rapid expansion of the fermentation sector, Wolff believes. The Remilk innovation team, he adds, is working with R&D teams at some of the world’s leading food companies. “What we’re seeing as a trend is the significant increase in the number of major manufacturers looking toward precision fermentation to power pipeline innovations,” he says. “As regulatory barriers fall, we’ll continue to see an increase in research, new product development, test and learn scenarios, and a variety of products from cheeses to dairy beverages hitting the market, and gradually transforming the dairy market as we know it.”

JENS BAYER, VICE PRESIDENT, MARKETING &  PRODUCT MANAGEMENT

The Pittsburg-based Scientific Bioprocessing Inc (SBI) contributes to modern fermentation methods by providing digital technologies to help researchers monitor bioprocessing activities. Its array of solutions, including sensors, actuators and software, has co-evolved with the food-tech sector.

When talking about food-tech sensors, many people visualize large-scale productions of high-grade proteins in massive fermenters. But a lot of time- and labor-intensive work has to take place before the final production step. The many optimization steps occur on a much smaller scale, for example in shake flasks.

“Although automation in bioreactors is the status quo, shake flask experiments are often still considered rudimentary, relying on manual sampling and thus missing crucial information,” says Jens Bayer, Vice President of Marketing & Product Management. “We seek to bridge the gap between these two vessel types. Our long-term vision is to provide bioreactor-like control and sensing options on a shake flask level but with much more simplicity and lower cost. We believe it will democratize fermentation and bioprocess development.”

Simplify your bioprocessing

SBI’s most recent breakthrough toward these goals was the launch of its DOTS platform. “The idea is to offer various sensors for several critical process parameters and automation technology, enabling real-time visualization and analysis for optimal bioprocess control,” Bayer reveals.

He contrasts the benefits of automation in fermentation with offline sampling, which he says is more complex and time-consuming, and also introduces risks of error and contamination. “This means critical information about your organism’s growth is being overlooked and could have a detrimental impact on the final product,” Bayer notes. “Automated online measurements, on the other hand, allow you to detect bioprocess changes in real-time. Although a lot of steps can already be automatized, there is still ground to cover.”

Although automation in bioreactors is the status quo, shake flask experiments are often still considered rudimentary, relying on manual sampling and thus missing crucial information

In terms of automation, certain vessel types, usually the larger ones, are often prioritized before others. That forces scientists to either accept the involvement of manual sampling or drives them to use larger-scale vessel type, such as bioreactors, even though other set-ups might fit the nature of their experiments better. “Companies must consider the entire lab, beginning with the smallest reaction vessels. Only then can a proper design of experiments be made without compromises.”

It’s also essential to be on guard for the drastic changes in conditions that can occur when bioprocesses are scaled up, Bayer cautions. It leaves the outcome unpredictable and, in many cases, the results will be disappointing. “Cultures have different nutrient or oxygen availabilities in larger scales and often different mixing or substrate feeding protocols are used compared to smaller scales. It’s vital to collect as much data as possible about the bioprocess before moving to large-scale fermenters. Mimicking the final bioprocess in smaller scales can avoid unpleasant surprises.”

SBI added important new expertise to its portfolio of technologies in 2021 when it acquired the German technology developer, aquila biolabs – a start-up whose innovations were a perfect synergy with SBI’s approach. Advanced solutions developed by aquila included its Cell Growth Quantifier (CGC), a smart sensor-based technology offering non-invasive online monitoring of biomass in shake flasks and other cultivation vessels. The company also brought with it CGQ BioR, which powers non-invasive, online monitoring of biomass in glass and single-use plastic bioreactors using backscatter measurements through the bioreactor wall. Meanwhile, SBI has also benefited from aquila’s Liquid Injection System (LIS) which enabled automated feeding of shake flasks for the first time.

Looking ahead, one of SBI’s main goals is to enlarge its DOTS platform. Bayer says they will keep two things in mind. “The first is to keep it simple,” he says. “The second is to keep the platform open for integration. We are focused on enabling scientists to have the best sensing experience possible. It is all about delivering the shortest time to value for customers, and giving them the flexibility to generate and structure their data.”

JULIE POST-SMITH, CHIEF OF COMMERCIAL, SUPERBREWED FOOD

Delaware, Minnesota-based biomass fermentation company, Superbrewed Food, is anticipating product launches toward the end of 2023 with multiple partners, including Bel Brands, the global player famous for best-selling cheese snacks such as The Laughing Cow, Boursin and Babybel.

Superbrewed Postbiotic Cultured Protein is made through the anaerobic fermentation of certain microflora found in nature, which converts plant fibers into protein, according to Julie Post-Smith, Chief of Commercial at the company. Pilot volumes are shipping from Superbrewed Food’s Delaware facility today.

“The alternative protein space is filled with challenges related to functionality and nutrition, but Superbrewed Food has developed a solution to both categories of challenges,” she says.

The company has partnered with the Bel Group to create cheese using Postbiotic Cultured Protein. “Their intention is to develop a full range of cheeses using the ingredient,” reveals Post-Smith. “When the agreement was signed last year after a screening process, Bel Group Research and Application Director, Anne Pitkowski, said she considered Superbrewed Food the ‘best in class startup’ in the field.”

As well as the agreement with Bel Brands, Superbrewed Food has and is developing additional deals with several other CPG companies in categories from dairy-free cheeses to yogurt, baked goods, and beverages.

Gorillas in the mix

Superbrewed Food was created in 2012 by Dr Bryan Tracy, who began wondering why some of the largest, strongest animals were herbivores, including silverback gorillas, which have the strength of 20 humans. Tracy and his R&D team found the answer in their digestive system, whose microbiomes are perfectly attuned to anaerobically ferment plant fibers into energy and protein.

The alternative protein space is filled with challenges related to functionality and nutrition, but Superbrewed Food has developed a solution to both categories of challenges

“From this original discovery, the team of researchers has de-risked the process of producing its postbiotic protein and it is now able to be produced at scale,” Post-Smith says. Superbrewed Food has developed a method of producing a natural, 85%-complete protein, which it claims is the highest concentration found in a single microbe. “The resulting Postbiotic Protein is better in nutrition than other alt proteins, providing high levels of vitamins, including B12, folic acid, biotin, riboflavin and pyridoxine, as well as minerals such as iron, zinc, selenium, molybdenum and phosphorus,” Post-Smith suggests. “Furthermore, it has a neutral taste, natural white color, and temperature stability, making it functionally superior to other alternative proteins.”

Superbrewed Food has identified a two-pronged production strategy. “To meet immediate demand, we are partnering for manufacturing. Originally, Superbrewed Food had planned to build out a facility in Minnesota to supplement fermentation capacity for start-ups looking to scale up operations commercially. That plan is second to the partner production plan.”

In May last year, a milestone was achieved when it received self-GRAS (Generally Recognized as Safe) status for its protein, signaling that an independent panel had accepted it posed little, or no, risk as an allergen. “It was the first microbiome protein to get approval by the FDA,” Post-Smith says.

Another step forward came in December 2022 when Superbrewed Food joined the San Francisco-based innovation platform, MISTA. The company is actively involved in the MISTA ecosystem and sharing its microflora protein technology to many other companies. The collaboration, Post-Smith concludes, will enable the company to start thinking more globally.

This article is republished from the April/May 2023 edition of Protein Production Technology International, the world's leading resource for the alternative proteins industry. If you wish to subscribe to future issues free of charge, please click here