Industrial fermentation

Technology
Technology details
Name:	Industrial fermentation
Category:	Conversion (Biochemical processes and technologies)
Feedstock:	Garden and park waste, food waste
Product:	Biomass, bioproducts (e.g., enzymes, biopolymers, organic acids, alcohols)

Industrial fermentation is a biotechnological process which uses microorganisms (genetically modified or not), in particular bacteria, yeasts, fungi or algae, to make useful products. The cells are real "cell factories" for the industrial conversion of a wide range of renewable feedstocks into bulk chemicals, fine chemicals, platform chemicals, pharmaceutical ingredients, bio-fuels, bio-plastics, etc. It is a multidisciplinary technology and includes the integrated application of disciplines such as biochemistry, microbiology, molecular genetics and process technology to develop useful processes and products.

Feedstock

Composition and origin

Depending on the type of microorganisms and its genetic modifications, a various range of feedstocks can be used. The most commonly used feedstocks are listed below:

Lignocellulose and cellulose

Lignocellulose is present in garden and park waste. Cellulose is present in food waste such as fruit and vegetable waste. Via hydrolysis, which is usually performed through enzymatic or thermal treatment, fermentable sugars can be obtained from lignocellulose and cellulose.

Starch

Starch is present in food waste such as potatoes, corn, wheat or cassava. Starch can directly be utilized by amylase-producing microorganisms, particularly filamentous fungi. However, to allow its use in a wider range of fermentations, starch is usually converted into glucose or dextrins by enzymatic hydrolysis.

Oils and Fat

Oils and fats are present in food waste such as gravy, used cooking oil and grease. They can directly be used as fermentation substrate. As they are not water soluble, extensive mixing is required to allow a good contact between the liquid droplets and the fermentation water phase.

Dairy waste

Whey, the liquid by-product of cheese manufacturing, is used as a source of fermentable carbohydrate and nitrogen.

Sugars

Sugar-rich waste streams can be derived from food industry waste, e.g., from the candy industry.

Pre-treatment

Depending on the type of feedstock and its purity, specific pre-treatment technologies are required to provide fermentable substrates to the microorganisms. Generally, this involves a size reduction step, after which the milled biomass can be processed to separate the desired substrate by e.g., centrifugation, filtration, evaporation or crystallization.

In addition, it should be taken into account that some of the above mentioned feedstocks only provide the carbon source (which compose about 50% of the weight of most microorganisms), in that case also other nutrients such as nitrogen, phosphate and potassium need to be added.

Process and technologies

Microorganisms

Microorganisms used in industrial fermentations include: bacteria, yeast, fungi or algae. In practice, these are well-known, productive and harmless (GRAS - Generally Regarded As Safe) production organisms, equipped with the new genetic information, that are used to produce the desired products in high yield and efficiency. A major advantage is that these often genetically modified microorganisms do their work under controlled conditions in a fermenter or bio-reactor, carefully contained and separated from the outside world (contained environment). They cannot escape from the factory so that ecological problems or concerns regarding the release of genetically modified organisms in the environment are avoided.

Schematic representation of an industrial fermentation bioreactor

Equipment

A typical industrial fermenter consists of an CSTR equipped with:

• an aeration and agitation system: to provide good mixing and availability of oxygen for the cell culture
• a temperature and pH control system: to assure optimal conditions for growth or production
• a foam control system: to avoid excessive foam formation
• sampling ports
• addition ports
• a cleaning and sterilization system: to avoid contamination with other, undesired microorganism

Operating conditions

As it involves living organisms, a fermentation process is typically conducted under mild conditions (pH and temperature). As a result, the energy consumption is relatively low as well as the capital and operating costs. However, fermentation technologies are complex and sensitive requiring careful control of quality and safety of the raw materials, process parameters, contamination, etc.

Industrial fermentations may be carried out as batch, fed-batch, or continuous culture systems. Batch and fed-batch operations are quite common, continuous fermentations being relatively rare .

Scale-up of industrial fermentations

Typically, a pure starter culture (or seed), maintained under carefully controlled conditions, is used to inoculate sterile petri dishes or liquid medium in the shake flasks. After sufficient growth, the preculture is used to inoculate the seed fermenter. Because industrial fermentations tend to be large (typically 1–250 m³), the inoculum is built up through several successively larger stages, to 5–10% of the working volume of the production fermenter. However, scale-up of a fermentation process is not straightforward as an increase in fermenter size affects the various process parameters in different ways. Therefore, ample expertise is required to find a compromise between all process parameters.

Products

Depending on the type of microorganisms and its genetic modifications, a range of products can be synthesized. The most common products are listed and divided over two categories: (1) biomass, (2) bioproducts. In case of the latter, some products require complex genetic modifications.

Biomass

• Single Cell Protein
• Single Cell Oil
• Baker's yeast
• Lactic acid bacteria

Bioproducts

Enzymes

• Proteases
• Lipases
• Amylases
• Cellulases
• Peroxidases

Biopolymers

• Poly-hydroxyalkanoates (PHA)
• Polysaccharides: xanthan gum, dextran

Organic acids

• Acetic acid
• Lactic acid

• Citric acid
• Tartaric acid
• Fumaric acid

Alcohols

• Ethanol
• Butanol
• Glycerol
• Butanediol

Solvents

• Acetone

Pharmaceuticals

• Vitamins: vitamin C, B2, B12 ...
• Antibiotics: aminoglycosides, penicillins, cephalosporins, tetracyclines ...
• Hormones

Biocolorants

• cartenoids
• astaxanthins

Biosurfactants and bioemulsifiers

• glycolipids
• rhamnolipids

Amino-acids

• monosodium glutamate (MSG)
• Lysine
• Tryptophan
• Phenylalanine

Post-treatment

The first step in the post-treatment of fermentation broth cultures, also known as downstream processing (DSP), is to remove the cells from the medium. This is typically performed by a solid-liquid separation technology, such, as centrifugation or membrane filtration. Each fraction can then undergo further processing, depending on whether the product is the biomass itself or an intra- or extracellular product. While intracellular products require cell disruption to release the products, extracellular products are solubilized in the depleted fermentation medium. Cell disruption techniques can be divided into mechanical methods (f.e. homogenisation, grinding, sonication, microwave treatment, steam explosion) and non-mechanical methods (f.e. osmotic or temperature shock, enzymatic destruction). To further purify and concentrate the products several methods can be used including chromatography, solvent extraction, crystallization, distillation, drying etc. The choice of purification technology is depending on the characteristics of the desired products.

Technology providers

Technology comparison

Company name	Country	Technology category	Technology name	TRL	Capacity [kg/h]	Feedstock: Food waste	Feedstock: Garden & park waste
Company 1	[Country HQ location]	[Technology category (if different sub-categories are defined this has to be specified here, the available categories can be found on each technology page under the chapter Process and technologies)]	[Technology name (the "branded name" or the usual naming from company side)]	[4-9]	[numeric value]	●	●
Company 2	[Country HQ location]	[(if different sub-categories are defined this has to be specified here, the available categories can be found on each technology page under the chapter Process and technologies)]	[Technology name (the "branded name" or the usual naming from company side)]	[4-9]	[numeric value]	●	●
PERSEO Biotechnology SL	Spain		PERSEO Bioethanol (R)	7-8	1000	●	●

Amphi-Star

Industrial fermentation provider

General information
Company:	AmphiStar
Country:	Belgium
Contact:	info@amphistar.be
Webpage:	https://www.amphistar.be
Technology and process details
Technology name:	BioSurf Biosurfactant Technology Platform	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	1-7	Capacity:	kg·h-1
Aeration:	Yes	Agitator:	Rushton
Biosafety level:	1	Controlled parameters:	Temperature, pH, Oxygen, Stirring speed, feed rates, etc.
Microorganism:	Starmerella bombicola, Candida kuoi, Rhodotorula bogoriensis, etc. Open for collaboration on any BSL-1 biosurfactant producing strain	Reactor material:	Glass or stainless steel
		Other:
Feedstock and product details
Feedstock:	Vegetable oils and sugars from biomass	Product:	Biosurfactants e.g. glycolipids such as sophorolipids

AmphiStar has developed a proprietary technology platform for the cost-efficient and ecological production of biosurfactants. We are a founders-led spin-off company established in July 2021 that is the result of 15 years joint development between Ghent University (Inbio.be) and the Bio Base Europe Pilot Plant. We derisk the early development stage for biosurfactant production, guide and support technology transfer to industrial manufacturers and collaborate intensely for further development and improvement of the licensed technology.

Our technology platform is initially based on the fermentative production with the yeast Starmerella bombicola, producing many different biosurfactants at a high volumetric productivity. Our biosurfactants are made from sustainable, renewable feedstocks and waste streams. Microbial fermentation is a clean production technology that is safe for people and the environment. Our biosurfactants are environmentally friendly, palm oil-free, sulfate-free, mild, non-toxic and non-irritant.

Sophie's BioNutrients

Industrial fermentation provider

General information
Company:	Sophie's BioNutrients
Country:
Contact:
Webpage:
Technology and process details
Technology name:		Technology category:	Conversion (Biochemical processes and technologies)
TRL:		Capacity:	kg·h-1
Aeration:		Agitator:
Biosafety level:		Controlled parameters:
Microorganism:		Reactor material:
		Other:
Feedstock and product details
Feedstock:		Product:

Avecom

Industrial fermentation provider

General information
Company:	Avecom
Country:	Belgium
Contact:	sales@avecom.be
Webpage:	https://www.avecom.be
Technology and process details
Technology name:	PROMIC	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	4-7	Capacity:	kg·h-1
Aeration:		Agitator:
Biosafety level:		Controlled parameters:
Microorganism:		Reactor material:
		Other:
Feedstock and product details
Feedstock:	Residual side streams and co-products from the food industry	Product:	Single Cell Protein, PHB-rich biomass

Avecom has developed its PROMIC biomass fermentation platform for the efficient conversion of industrial and agricultural residual side streams and co-products towards high-value single cell proteins.

Cetaqua Galicia

Industrial fermentation provider

General information
Company:	Cetaqua Galicia
Country:	Spain
Contact:	anton.taboada@cetaqua.com
Webpage:	https://www.cetaqua.com/
Technology and process details
Technology name:	TCP (The Carboxylic Platform)	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	7	Capacity:	0.43 - 0.63 kg·h-1
Aeration:	No (Anaerobic technology)	Agitator:	Pitched blade agitator
Biosafety level:	1	Controlled parameters:	Temperature, pH, stirring speed, feed rates and moisture.
Microorganism:	Anaerobic open mixed culture (uncontrolled)	Reactor material:	Stainless steel
		Other:
Feedstock and product details
Feedstock:	Seawage sludge or urban biowaste	Product:	Acetic acid, propionic acid, butyric acid and valeric acid.

Cetaqua Galicia is a public-private research centre founded in 2011 by Viaqua, the University of Santiago de Compostela (USC) and the Superior Council of Scientific Investigations (CESIC). Through our three lines of research, we have positioned ourselves as a benchmark centre, at regional, national and European level, in the application of scientific knowledge to the water cycle, especially the fields of wastewater treatment and production and recovery of high value-added by-products in waste water.

Within the biofactory line, the Volatile Fatty Acids (VFAs) production technology from urban sewage sludge and urban biowaste has been developed.  The technologies was validated on a laboratory scale in 2020, and two pilot prototypes were designed and built. The first one installed at Conservas Dardo to recover industrial wastewater from the canning industry, and the second at the Ourense wastewater treatment plant for the recovery of sludge from urban wastewater treatment plants. In addition, this project is currently preparing innovation proposals for the development of the corresponding patents for the VFA line.

Holiferm

Industrial fermentation provider

General information
Company:	Holiferm
Country:	United Kingdom
Contact:	Joana Pereira (info@holiferm.com)
Webpage:	https://holiferm.com/
Technology and process details
Technology name:	Fermentation intensification and in-line separation	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	1-9	Capacity:	Lab scale to pilot (600 L) kg·h-1
Aeration:	Aerobic, semi-anaerobic, anaerobic	Agitator:	Rushton
Biosafety level:	Up to biosafety level 2	Controlled parameters:	Temperature, pH, pressure, stirring rate, flows, dissolved oxygen, etc.
Microorganism:	Bacteria and yeast	Reactor material:	Glass (lab scale), stainless steel (pilot and commercial)
		Other:	https://www.linkedin.com/company/holiferm/
Feedstock and product details
Feedstock:	Vegetable oil and sugar streams	Product:	Biosurfactants (other biomolecules in the future)

Holiferm develops holistically improved fermentation technology and processes to deliver massive economic improvements, with biosurfactants being the initial focus. Holiferm’s patented integrated gravity separation and fermentation technology is a plug and play system that increases fermentation process productivity by three to four times, reducing production costs by more than 50% and enabling low cost production of biosurfactants: renewable, biodegradable and mild alternatives to the petrochemical derived surfactants used in household cleaning, laundry and personal care products. Holiferm is dedicated to the commercialisation of economic biosurfactant production processes, providing a complete platform technology for production, isolation and purification, enabling significant market disruption and growth.

MOA foodtech

Industrial fermentation provider

General information
Company:	MOA foodtech
Country:	Spain
Contact:
Webpage:
Technology and process details
Technology name:		Technology category:	Conversion (Biochemical processes and technologies)
TRL:		Capacity:	kg·h-1
Aeration:		Agitator:
Biosafety level:		Controlled parameters:
Microorganism:		Reactor material:
		Other:
Feedstock and product details
Feedstock:		Product:

Nosh.bio

Industrial fermentation provider

General information
Company:	Nosh.bio
Country:	Germany
Contact:	info@nosh.bio
Webpage:	https://www.nosh.bio/
Technology and process details
Technology name:	Sustainable functional ingredients for the food industry	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	5	Capacity:	Not disclosed kg·h-1
Aeration:	Yes	Agitator:	Pneumatically agitated (e.g., airlift)
Biosafety level:	1	Controlled parameters:	Not disclosed
Microorganism:	Filamentous fungi	Reactor material:	Not disclosed
		Other:
Feedstock and product details
Feedstock:	Several	Product:	High quality functional microbial biomass

NovelYeast bv

Industrial fermentation provider

General information
Company:	NovelYeast bv
Country:	Belgium
Contact:	johan.thevelein@novelyeast.com
Webpage:	https://www.linkedin.com/in/johan-thevelein-aab60a10/
Technology and process details
Technology name:	Yeast fermentation to biofuels and bio-based chemicals. Protein production	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	3-5	Capacity:	Lab-scale kg·h-1
Aeration:	Aerobic, semi-anaerobic	Agitator:	Shake flasks, static tubes with magnetic stirring
Biosafety level:	BSL-1	Controlled parameters:	Standard parameters
Microorganism:	Saccharomyces cerevisiae, other yeast species, Trichoderma	Reactor material:	Glass
		Other:	Construction of cell factories with recombinant DNA technology
Feedstock and product details
Feedstock:	1G and 2G feedstocks	Product:	Biofuels and bio-based chemicals, proteins, specialty sugars, specialty chemicals

NovelYeast bv was founded in 2019 by Prof. Johan Thevelein (KU Leuven and VIB) to continue his R&D activities after his retirement in 2020 as emeritus. The company focusses on the development and industrial implementation of yeast cell factories for the production of biofuels, bio-based chemicals as well as specialty sugars and ingredients with first- and second-generation feedstocks. It also develops cell factories for the production of specific proteins for food applications and enzymes for saccharification of lignocellulosic biomass. In addition, it uses yeast as a tool for biomedical and agroindustrial applications, including yeast probiotics and anti-cancer drugs selected by screening in yeast. NovelYeast has several R&D service collaborations with companies world-wide.

PERSEO Biotechnology SL

Industrial fermentation provider

General information
Company:	PERSEO Biotechnology SL
Country:	Spain
Contact:	informacion@perseobiotech.com
Webpage:	https://www.perseobiotech.com/
Technology and process details
Technology name:	PERSEO Bioethanol ®	Technology category:	Conversion (Biochemical processes and technologies)
TRL:	7 - 8	Capacity:	1000 kg·h-1
Aeration:	If needed. Currently under anaerobic conditions.	Agitator:	Vertical stirrers
Biosafety level:	High, no dangerous biological material used.	Controlled parameters:	Temperature, pH, pressure, stirring rate, flows, dissolved oxygen.
Microorganism:	Yeasts and bacteria	Reactor material:	Stainless steel
		Other:	not applicable
Feedstock and product details
Feedstock:	Biodegradable waste (OFMSW, agro-industrial waste, cellulosic waste, etc.)	Product:	Advanced Bioethanol + CO2+ valuable organic byproduct

PERSEO Biotechnology SL is a Spanish SME with track experience and know-how in the development of biotechnological processes, which range from the development phase at the laboratory level to the industrial upscaling of the process and its demonstration. Likewise, PERSEO Biotechnology offers complementary services to assess the feasibility and the scalability of the biotechnological processes.

PERSEO Biorefinery has its own laboratories and a versatile semi-industrial plant (L’Alcudia, Valencia, Spain) with a treatment capacity up to 25 tons / day of organic waste whose objective is to develop, test and validate biotechnological processes to generate bioproducts and bioenergy, integrating all R&D services for the global recovery of organic waste.

PERSEO Bioethanol® (http://www.perseobiotech.com) is a patented and innovative technology to convert organic waste, such as biodegradable municipal solid waste, horticultural waste, agro-industrial waste, HORECA channel or paper and cardboard, mainly into advanced bioethanol, to be used as liquid biofuel or as raw material for the chemical industry, and in other bioproducts with high potential in the chemical industry (bioproducts from the fermentation of sugars, biosurfactants, biofertilizers or in biomethane by anaerobic digestion).

PERSEO Bioethanol® is a patented biotechnological process compatible with current existing waste treatment plants, under the concept of an integrated biorefinery. It is adaptable to each process and to the needs of each client. The process can be integrated as a previous recovery stage in existing plants, including incineration, anaerobic digestion or composting, increasing the value chain of waste treatment and significantly improving the economic and environmental results of waste management.

POLYFOODS

Industrial fermentation provider

General information
Company:
Country:
Contact:
Webpage:
Technology and process details
Technology name:		Technology category:	Conversion (Biochemical processes and technologies)
TRL:		Capacity:	kg·h-1
Aeration:		Agitator:
Biosafety level:		Controlled parameters:
Microorganism:		Reactor material:
		Other:
Feedstock and product details
Feedstock:		Product:

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