The future in animal feed - Agmondo...M. capsulatus Single cell protein The substrates for making UniProtein® are: a nitrogen source (ammonia), an energy source (methane or methanol),

The future in animal feedFeeding the world an increasingly scarce, though essential nutrient

Table of contents

About Unibio

The concept

The U-Loop® technology

UniProtein®

UniProtein® in the protein demand chain

Environmentally friendly production

Chemical composition

Seven reasons to use UniProtein® in animal diets

UniProtein® in pig diets

UniProtein® in broiler diets

UniProtein® in mink diets

UniProtein® in blue fox diets

UniProtein® in aqua diets

The proper use of UniProtein® in animal diets

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Unibio is a Danish biotechnology company with a core focus on fermentation technologies, allowing a highly scalable production of bacterial protein meal. Since its foundation the company’s goal has been to supply the world with innovative sustainable solutions to overcome the food challenge faced by the growing global population. With the use of Unibio’s biotechnological innovations the company is able to transform processes that occur in nature every day into a bioindustrial setting and produce healthy sustainable protein for animals in an eco-friendly way.

Unibio owns the intellectual property rights to a unique fermentation technology – the U-Loop® technology – producing a high-value bacterial protein product, UniProtein®. This novel protein source disrupts the traditional food value chain while mitigating the traditional challenges by decoupling protein production from the fluctuating agriculture and stressed fishing industries. Crop land can thus be used to produce food for humans instead of feed for animals.

UniProtein® is a sustainable solution to the world’s protein-scarcity challenge.

About Unibio

4

The concept

Cooling water

Fed into the fermentor:-Air-Methane-Minerals-Oxygen-Catalysts

Viscous protein broth is sprayed out

Propeller pump keeps the liquid moving

Temperature is held at 45”C

Hot air from the bottom of the fermentor dries the broth, turns itinto protein granule and makes it fall to the bottom

Protein granules are excellent animal feed

Separator separates water from the protein broth

Drying tower

Carbon dioxide and unused nitrogen from the air are let out

Separator

Process:Bacteria eat methane, and the liquid is transformed to protein broth

This process occurs in nature every day e.g. in lakes

U-Loop® fermentor

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The U-Loop® technology

The U-Loop® technology provides optimal growth conditions for the bacteria. The tailor-made technology speeds up the natural process significantly while allowing the execution of the procedure in a controlled and monitored environment. Unibio has developed a vertical fermentor design that can handle a large volumetric gas fraction, significantly improving the average productivity. The technology has been developed in cooperation with the Technical University of Denmark (DTU) after more than 30 years of R&D and is patented under process patent No PA199900690.

The U-Loop® fermentor allows for a much more energy-efficient transport of nutrients from gas phase into liquid phase, at a much higher rate compared to conventional bioreactors.

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UniProtein®

is well suited as a protein feed material for livestockis a free-flowing reddish-brown granule with a particle size of 150-200μmhas a consistent chemical composition and protein contenthas a long shelf lifedoes not contain genetically modified microorganisms (non-GMO)is a non-polluting product as it is produced by a microbial culture with methaneis free from toxins, dioxin and heavy metals due to the controlled production process and thefact that all minerals used are food gradeaddresses the world’s growing need for animal protein, which will continue to increasein line with population growthis produced without using any pesticides or fertilizersis weather-independentproduction does not require any arable land and utilizes a minimum amount of water

UniProtein® was approved for animal nutrition in the EU in July 1995 according to Commission Directive 95/33/EC. Under EU Commission Regulation No 68/2013 of 16 January 2013, UniProtein® is listed on the catalogue of feed materials (under 12.1.2: Protein from Methylococcus capsulatus (Bath), Alcaligenes acidovorans, Bacillus brevis and Bacillus firmus).

UniProtein® is a brand-specific bacterial protein meal. It is a protein-rich biomass produced by a microbial culture with methane as the sole carbon and energy source.

UniProtein® contains the following four heat-killed bacteria:

The production process always results in a uniform product.

M. capsulatus Single cell protein

The substrates for making UniProtein® are: a nitrogen source (ammonia), an energy source (methane or methanol), minerals and oxygen.

Methylococcus capsulatus (90%) (NCIMB strain 11132)Ralstonia sp. (8%) (NCIMB strain 13287, formerly known as Alcaligenes acidovorans)Aneurinibacillus danicus (1%) (NCIMB strain 13288, formerly known as Bacillus brevis)Bacillus firmus (1%) (NCIMB strain 13288)

Minerals

Ammonia

Oxygen

Methane

UniProtein®

7

UniProtein® in the protein demand chain

Protein is the building block of all life and essential for the growth of cells and tissue repair.The growing meat consumption triggers an increasing demand for animal husbandry and aquaculture which in turn require more animal feed and consequently sufficient crude protein.Unibio offers a unique alternative to existing sources of protein for animal feed by the use of an abundant raw material, methane.

Fish

Chicken

Eggs and dairy

Cattle

Pigs

Fish

Chicken

Pets

Compound feed

Soybean meal

Fishmeal

Other

By-products

Agriculture

Fishing

Rendering

Methane

Pet food

Pork

Beef

MeatAnimal

husbandryand aquaculture

Animal feed Crude protein Raw materials

UniProtein®

Imagine a future where the world’s protein supply is partly covered by a technology enabling methane to be turned into protein by bacterial fermentation

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Environmentally friendly production

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1Global land useLand use varies significantly depending on the geographical location where the crops are cultivated.

Land use1 of typically used feed ingredients in animal diets

Total water1 footprint of typically used ingredients in animal diets

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m2 / t

1Global total water consumption. 2The total water consumption (4.6 m³/t) is equal to blue water consumption during the production of UniProtein®. Total water consumption includes green, blue and grey water consumption.

Methane O2, phosphoric acid, ammonia

Main inputs By-products

Clean waterand limited CO2

Components of the production process

The production of UniProtein® results in:

Minimal use of water (which is also a by-product)12 x output per hectare of arable land compared to soybean mealNo need for pesticides

Uniform qualityStorage stabilityRegular supply at fixed prices

High % of crude protein and targeted amino acidsEnvironmentally friendly productModifiability to different animal groups

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“The simulation results from a leading feed optimization software manufacturer showed that when UniProtein® was added by up to 5% of the total piglet feed weight, 6.5% less soybean meal was required. This alone resulted in a reduction of 250 kg CO2 eq. / tonne of feed. This equals to driving a car from Southern Denmark to Venice, Italy, for just one tonne of feed”. Lorie Hamelin, October 2015Senior Scientist and PhD, Institute of Soil Science and Plant Cultivation, Department of Bioeconomy and Systems Analysis, Poland

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Compound(dry matter - DM¹)

Meanvalues

Crude protein (N x 6.25) 72.9%

Crude fat 9.1%Ash 550 8.6%N-free extracts 7.6%Nucleic acids

Ribonucleic acid (RNA) 6.7%Deoxyribonucleic acid (DNA) 2.3%

Sugars(after acid digestion)

Glucose 4.5%Other monosaccharides

Seven reasons to use UniProtein® in animal diets

2. Ideal amino acid profileUniProtein® has an ideal amino acid (AA) profile. UniProtein® has a higher concentration (% of CP) of valine, tryptophan, isoleucine and leucine compared to fishmeal. In the rest of the essential AAs there are minor differences between UniProtein® and average data for fishmeal and SBM.

AA = Amino acidCP = Crude proteinDM = Dry matterSBM = Soybean meal1 The low lysine content of UniProtein® can easily be addressed by adding feed-grade lysine to complete diets

Source of values for fishmeal and SBM: http://www.feedipedia.org

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Soybean meal, CP 53.5 (% DM) (dehulled)Fishmeal, CP 70.6 (% DM)UniProtein®, CP 72.9 (% DM)

ValTyrTrpThrSerProPheMetLys1LeuIleHisGlyGluCysAspArgAla

The content of AAs (% of CP) of UniProtein® in comparison with fishmeal and SBM

UniProtein®, CP 72.9 (% DM) Fishmeal, CP 70.6 (% DM) SBM, CP 53.5 (% DM) (dehulled)

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Ala Arg Asp Cys Glu Gly His Ile Leu Lys1 Met Phe Pro Ser Thr Trp Tyr Val

DM = Dry matter

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1. Competitive protein contentThe crude protein (CP) content of UniProtein® is superior to soybean meal (SBM) and comparable to fishmeal.

The crude protein content of UniProtein® in comparison to fishmeal (60-68% CP as fed) or SBM.

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3. Low ash and mineral contentThe ash and mineral content of UniProtein®, particularly phosphorus and calcium, is low compared to fishmeal. This makes UniProtein® the ideal protein compound to incorporate into animal feed when a high protein content and a low mineral and ash content are required.

4. Valuable fatty acid profileThe fatty acid profile of UniProtein® consists primarily of saturated C16:0 and monounsaturated C16:1 fatty acids representing approx. 85% of total fatty acids (TFA). Results of experiments indicate that the fatty acid profile of UniProtein® results in good oxidative stability of frozen pork and chicken meat.

5. UniProtein® is rich in B vitaminsB vitamins are water-soluble and are not stored in the tissues in appreciable amounts. Their regular supply is essential.

6. Neutral flavour UniProtein® has a neutral taste. The palatability of diets including UniProtein® has been shown to be as well accepted as that of diets including typical protein sources.

7. Utilization and digestibilityThe digestibility of protein and AAs of UniProtein® diets and the retention of nitrogen (N) of animals on UniProtein® diets in the experiments have been comparable to the control diets. The main protein source in the control diets has been either fishmeal or SBM.

The ash, calcium and phosphorus contents (% of DM) of UniProtein® in comparison with conventional-commodity fishmeal with a CP content of 70.6% of DM.

Source of values for fishmeal and SBM: http://www.feedipedia.org

DM = Dry matter

Ash, calcium and phosphorus (% of DM)

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Impact of increased inclusion of UniProtein® on the retention of nitrogen (N) and heat energy

PerformanceThe impact of increased inclusion of UniProtein® on pig performance

Nutrient metabolism

Average daily gain1, g

N Retention, g/kg 0.75 per day

N = Nitrogen

Heat energy, kJ/kg 0.75 per day

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1 from weanling (11.4 kg) to slaughter (107.2 kg)

The average daily gain (ADG), average daily feed intake and feed conversion ratio (FCR) were similar among diets including UniProtein® from 0% to 10%.

Increased levels of UniProtein® were balanced by a reduction of protein derived from SBM.

Source: Øverland et al. 2004, Ital. J. Anim. Sci. 3:323-336

UniProtein® incorporated by up to 15% into pig diets maintained the good N retention and showed no adverse effects on heat energy. Increased levels of UniProtein® were balanced by a reduction of protein derived from SBM.Source: Hellwing et al. 2007, Animal 1:45-54

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Meat qualityThe impact of increased inclusion of UniProtein® on the quality of frozen pork

PUFA, % of TFA in backfat

Fat firmness1

Rancid taste1

PUFA = Polyunsaturated fatty acidsTFA = Total fatty acids

1 On a scale from 1 (poorest) to 9 (best)

1 On a scale from 1 (poorest) to 15 (best)

Increased levels of UniProtein® in the diets decreased the ratio of polyunsaturated fatty acids (PUFA) to total fatty acids (TFA) in backfat and led to better oxidative stability of frozen pork as shown by the improved rancid taste.Increased levels of UniProtein® were balanced by a reduction of protein derived from SBM.

Source: Øverland et al. 2005, Meat Sci. 71:719–729

The increased levels of UniProtein® in the diets improved fat firmness in a linear manner.Increased levels of UniProtein® were balanced by a reduction of protein derived from SBM.

Source: Øverland et al. 2004, Ital. J. Anim. Sci. 3:323-336

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The impact of increased inclusion of UniProtein® on broiler performance when replacing SBM Performance

The impact of increased inclusion of UniProtein® on the amino acid digestibility of diets when replacing SBMDigestibility

AA = Amino acidAID = Apparent ileal digestibility1 Measured at day 32

The apparent ileal digestibilities (AID) of amino acids were unaffected or improved when UniProtein® replaced SBM in the diets.

Source: Schøyen et al. 2007, Arch. Anim. Nutr. 61:276-291

The body weight and feed intake were similar among the diets including UniProtein® from 0% to 12%. The FCR was statistically similar among the diets. However, UniProtein® inclusion resulted in the numerical improvement of FCR compared to the control diet.

Source: Øverland et al. 2010, Br. Poult. Sci. 51:686-695

UniProtein® 12%

UniProtein® 8%

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ValTrpThrPheMetLysLeuIleHisArg

Body weight1, g

FCR = Feed conversion ratio1 day 0 - day 32

1 day 32

FCR1, g feed/g weight gain

Coefficients of AID of essential AAs in diets1

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Meat qualityThe impact of increased inclusion of UniProtein® on the quality of chicken meat when replacing SBM

UniProtein® decreased the ratio of PUFA to TFA in chicken fat, reduced the level of oxidative products in frozen meat and had no adverse effects on the several sensory quality parameters related to odour and flavour.

Source: Øverland et al. 2011, Poult. Sci. 90:201-210.

1 in the thigh meat after 6 months of storage at −18°C2µg/g ×103µg/g

PUFA = Polyunsaturated fatty acidsTFA = Total fatty acids

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PUFA, % of TFA Oxidative products1

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PerformanceThe impact of increased inclusion of UniProtein® on broiler performance when replacing fishmeal

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Body weight, feed intake and FCR were similar among the diets including UniProtein® from 0% to 6%.Source: Schøyen et al. 2007, Arch. Anim. Nutr. 61:276-291

DigestibilityThe impact of increased inclusion of UniProtein® on the amino acid digestibility of diets when replacing fishmeal

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AID = Apparent ileal digestibility AA = Amino acid 1 Measured at day 32

The AIDs of amino acids were unaffected when UniProtein® replaced fishmeal in the diets.


Body weight1, g FCR1, kg feed/kg body weight gain

Coefficients of AID of essential AAs in the diets1

1 day 32 FCR = Feed conversion ratio 1 day 0 - day 32

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The impact of increased inclusion of UniProtein® on the protein and energy metabolism


Nutrient metabolism

The values of retained N and heat energy were similar among the diets including UniProtein® from 0% to 6% of diet. Increased levels of UniProtein® were balanced by a reduction of protein derived from fishmeal.

Source: Hellwing et al. 2006, Arch. Anim. Nutr. 60:365-281

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N retention, g /kg 0.75 per day

N = Nitrogen

Heat energy, kJ/kg 0.75 per day

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Replacing fishmeal by UniProtein® in the diet improved the sensory quality of frozen-stored chicken meat as shown by the less intense off-odour and off-flavour and increased the juiciness of frozen meat. The off-odour and off-flavour decreased, and juiciness increased numerically when UniProtein® replaced SBM in the diet.


1 On a scale from 1 (no intensity) to 9 (high intensity)2175 freeze-stored days

1 On a scale from 1 (no intensity) to 9 (high intensity)2 175 freeze-stored days

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Meat qualityThe impact of increased inclusion of UniProtein® on the quality of chicken meat

Juiciness1,2

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Off-odour Off-flavour

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The body weight gain, feed intake and FCR were similar among diets including UniProtein® by 0% to 8%.Increased levels of UniProtein® were balanced by a reduction of protein derived from fishmeal.

Source: Ahlstrøm et al. 2006, J. Anim. Feed Sci. 15:491-504

UniProtein® incorporated by up to 8% in the diets improved the total impression of fur quality characteristics compared with the control diet.Increased levels of UniProtein® were balanced by a reduction in protein derived from fishmeal.


1On a scale from 1 (poorest) to 9 (best)1from 29 June to 26 November (a total of 121 feeding days)

Body weight gain1, g

Performance and fur qualityThe impact of increased inclusion of UniProtein® on mink performance

Total impression of fur quality characteristics1

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Nutrient metabolism and digestibilityThe impact of increased inclusion of UniProtein® on N retention and the digestibility of N and AAs in the diet

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Coefficients of ATTD of AAs in the diet

N digestibility, %

N = Nitrogen

AA = Amino acidATTD = Apparent total tract digestibility

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UniProtein® incorporated by up to 8% into male mink diets maintained the good retention and digestibility of N.Increased levels of UniProtein® were balanced by a reduction in protein derived from fishmeal.


UniProtein® incorporated by up to 14.7% into mink diets maintained the good apparent total tract digestibility (ATTD) of main nutrients and AAs.Increased levels of UniProtein® were balanced by a reduction in protein derived from fishmeal.

Source: Øverland et al. 2006, Anim. Feed Sci. Tech. 129:237-2510,0

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UniProtein® 14.7%

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1.4 100

1.280

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UniProtein® in blue fox diets

Nutrient metabolism and digestibilityThe impact of increased inclusion of UniProtein® on the production performance and diet digestibility in blue foxes.

The study was conducted to investigate the potential use of UniProtein® in diets of dogs using blue foxes as a model.

Final body weight1, kgATTD of main nutrients in the diet, %

Final body weight, overall gain and feed consumption were similar among diets including UniProtein® by 0% to 12%.

The ATTDs of main nutrients were similar among the diets including UniProtein® from 0% to 12%.Source: Skrede and Ahlstrøm 2002, J. Nutr. 132: 1668S–1669S

The ATTD of crude protein

The ATTD of crude fat

The ATTD of crude carbohydrate

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PerformanceThe impact of increased inclusion of UniProtein® on the performance of Atlantic salmon when replacing fishmeal


In a 12-month experiment the salmon (at age of 112 days) fed the diet containing 5% UniProtein® weighed significantly more than salmon fed the fishmeal-based control diet, while salmon fed the diets containing more UniProtein® reached the same weight as the control group.

The salmon (at age of 364 days) fed the diet containing 19.3% UniProtein® ended up with a higher final weight than salmon fed the fishmeal-based control diet or the diets including less UniProtein®. Specific growth rate and survival of fish fed up to 19.3% UniProtein® were similar among diets during the entire experiment.

Source: Storebakken et al. 2004, Aquaculture, 241:413-425

1day 112 1day 364

1day 52

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UniProtein® 18%

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40

50

UniProtein® 19.3%UniProtein® 9.9%UniProtein® 5%UniProtein® 0%

Body weight1, g

Final weight1, g Final weight1, g

UniProtein® improved the body weight of juvenile Atlantic salmon.

Source: Aas et al. 2006a, Aquaculture, 259:365-376

UniProtein®0%

UniProtein®0%

UniProtein®0%

UniProtein®5%

UniProtein®4.5%

UniProtein®5%

UniProtein®9.9%

UniProtein®9%

UniProtein®9.9%

UniProtein®19.3%

UniProtein®18%

UniProtein®27%

UniProtein®19.3%

4 50

340

230

1

20

10

0 0

370

360

350

340

320

330

310

300

23


PerformanceThe impact of increased inclusion of UniProtein® on the performance of rainbow trout when replacing fishmeal

The specific growth rate and feed intake of trout were similar among diets including UniProtein® from 0% to 18%. The FCR was similar between the control diet and the diet including 18% UniProtein®, while the FCR of trout on a diet including 9% UniProtein® outperformed the FCR of trout on the control diet or the diet containing 18% UniProtein®.

Source: Aas et al. 2006b, Aquaculture, 261:357-368

1day 76 FCR = feed conversion ratio1day 0 – day 76

FCR1, g weight gain/g feed Specific growth rate1

1,0

1,1

1,2


1,35

1,36

1,37

1,38

1,39

1,40

1,41

UniProtein® 18%UniProtein® 9%UniProtein® 0%UniProtein®0%

UniProtein®0%

UniProtein®9%

UniProtein®9%

UniProtein®18%

UniProtein®18%

1.411.2

1.40

1.39

1.381.1

1.36

1.37

1.351.0

24

Nutrient metabolism and digestibilityThe impact of increased inclusion of UniProtein® on the diet digestibility of rainbow trout


0

10

20

30

40

50

60

70

80

90

100


AID of N, % AID of energy, %

N retention (% digested)

0

10

20

30

40

50

60

70

80

90

100


0

10

20

30

40

50

60


The AIDs of N, energy and AAs in rainbow trout differed slightly among diets including UniProtein® by 0% to 18%.

The N retention in rainbow trout were similar among diets including UniProtein® by 0% to 18%.Increased levels of UniProtein® were balanced by a reduction in content of protein from fishmeal.

Source: Aas et al. 2006b, Aquaculture, 261:357-368

AID = Apparent ileal digestibility N = Nitrogen

AID = Apparent ileal digestibility

N = Nitrogen

UniProtein®0%

UniProtein®0%

UniProtein®0%

UniProtein®9%

UniProtein®9%

UniProtein®9%

UniProtein®18%

UniProtein®18%

UniProtein®18%

100 10090 9080 8070 70

50

50

5060

60

60

40

40

4030

30

3020

20

2010

10

100

0

0

25

Maximum inclusion levels of UniProtein® in animal diets based on initial studies

The proper use of UniProtein® in animal diets

1 Based on trials in broilers and considering the low protein content of layers’ diets compared to the diets of broilers2 Based on trials conducted in pigs from approx. 10 kg to 80 – 110 kg3 Based on trial in blue foxes

Species

Mink

Blue fox

Dog and cat3

Broiler

Pig2

Laying hen1

Inclusion level (% of diet)

8

6

3

8

12

12

Rainbow trout 20

Atlantic salmon 20

26

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Documents

The future in animal feed - Agmondo...M. capsulatus Single cell protein The substrates for making UniProtein® are: a nitrogen source (ammonia), an energy source (methane or methanol),