Chlamydomonas reinhardtii, a green algae for plant and animal proteins
By Xun Wang, President and CEO
BIOWC18 – Company & Technical Presentation - 19 July 2018 Our world is facing increasing demands for proteins
• Global population will grow from today’s 7.6 billion to 9.8 billion by 2050 • Global meat consumption is projected to increase by 76% • Future crop yields need to grow 32% faster than before • Meat production adds pressure to crops, water resources and the environment • Plant proteins impact on arable lands, lack of essential amino acids and taste
Start-up companies are building momentums in meat and dairy substitute innovations
Source: Maeve Henchion, 2017. Future Supply and Demands
2 Chlamydomonas reinhardtii
• Chlamydomonas reinhardtii is a single-cell green alga.
• Chlamydomonas species are distributed worldwide.
• Chlamydomonas reinhardtii has been a model organism, with a flexible suite of synthetic biology tools: an ideal host to produce mammalian proteins, monoclonal antibodies, vaccines, and hormones.
• Previously, Chlamydomonas lacked an economic large scale production process and food safety record.
3 For the first time, Triton has developed a commercial viable production process for Chlamydomonas reinhardtii
• Productivity: Increased from < 0.4 to > 20 grams/liter/day • Titers: Increased from < 2 grams/liter to > 120 grams/liter • Validations: Process validation by two independent contract manufactures in 200 liter tanks, and one in 3,000 liter tanks • Production costs: Estimated between $2.17 (by 2021) to $7.76 per kg (today) as dried powder when scaled up
4 Chlamydomonas (Chlamy) is rich in protein, omega oils, vitamin A, fiber, and essential amino acids
Fat Cholesterol Carb Fiber Protein** Vit A Ca++ Iron Ω-oil* daily daily daily daily daily daily value daily value daily value daily value grams value mg value grams value grams value grams value Wheat 1.4 2% 0 0% 71 25% 11 40% 4 7% 0% 3% 6% 0% Rice 1.4 2% 0 0% 80 28% 2.4 9% 6 11% 0% 1% 2% 0% Corn 3.9 5% 0 0% 77 27% 7.3 26% 7 13% 0% 1% 13% 0% Soybean 20 24% 0 0% 30 11% 9.3 34% 36 65% 0% 28% 78% 0% Beef*** 10 12% 90 30% 0 0% 0 0% 20 36% 0% 2% 20% 0% Cricket 15 18% 300 100% 9 3% 9 33% 25 45% 0% 24% 33% 0% Chlamy 7 8% 0 0% 34 12% 8 29% 48 86% 24% 4% 80% 847%
Serving size: 100 grams Omega-oil*: 250 to 500 mg of omega oil for healthy adult, 1 gram for coronary heart disease adult Protein** daily value: Adult men need about 56 grams a day. Adult women need about 46 grams a day Beef***: 90% lean ground beef. 69% water. Essential amino acids: Soybean = 47%. Chlamy = 58%.
5 Chlamydomonas is safe to eat: Triton has submitted a notification of GRAS to the FDA in April 2018
• Three in vitro assays were completed and showed no adverse or toxic effects o Mutagenicity (Ames Bacterial Reverse Mutation Assay) o Clastogenicity (Chromosomal Aberration Assay) o Genotoxicity (Mouse Micronucleus Assay)
• 28-day rat feeding trial was completed and showed no adverse or toxic effects even at the highest dose level
6 Chlamydomonas reinhardtii is becoming the newest member in nutritional food ingredients
Safe to eat Good taste Nutritious Two different colors Long shelf life Scalable production Environment friendly No pesticides No bacteria contaminations
7 Chlamydomonas was first introduced into restaurants in May 2018
8 Although a few other algae species are also safe to eat, Chlamydomonas reinhardtii is the only one with advanced proprietary synthetic biology tools for alternative milk and meat
Chlamydomonas Arthospira spp. Chlorella spp. Dunaliella spp. Haematococcus Nanochloropsis spp. spp. Advance Genetic Tools Scalable Production Mating/Breeding
Fermentation Capable GRAS
9 Leveraging on Chlamydomonas food applications, Triton will further differentiate itself through synthetic biology to enter three fast growing markets
Gut Health Infant Formula Meat Imitations
Triton’s • Base strain improves gut health • Like Osteopontin, many colostrum • Triton has colorless strain, Advantages • Many colostrum proteins proteins can maintain bioactivities ideal for leghemoglobin benefit gut health when synthesized in Chlamydomonas and myoglobin • No need to purify from the host • Triton has 14 colostrum proteins in the productions • Strain + colostrum proteins pipeline • Unlike other alternatives, build product series • Lead protein, Osteopontin, has been no purification is required, • Have patents on processes and successfully produced, purified, therefore much lower cost utilities to protect Triton validated for its biological activities to compete
10 Three independent animal trials have demonstrated wildtype Chlamydomonas supports gut health. Human trial is underway.
1. Mouse Trial - DSS challenge (14 days) • Reduces severity (41%) • Number of mice: 22 mice • Supports recovery of colitis • Dependent variables: Weight in mice (44%)
2. Piglet Trial – E.coli challenge (21 days) • Diarrhea incidence dropped • Number of piglets: 24 piglets from 35% to 24%, a • Dependent variables: Diarrhea reduction in the incidence incidence of diarrhea by 31%
3. Piglet Trial – Exposure to natural • Algae performed as well as disease (14 days) Zinc and Aureomycin • Number of piglets: 118 piglets • Algae reduced the • Dependent variables: Diarrhea incidence of diarrhea over incidence a control diet by a statistically significant 63%
confidential 11 Triton has developed a robust pipeline of milk proteins
Osteopontin (OPN), future • Like osteopontin (OPN), ingredient in infant formula many colostrum proteins can maintain bioactivities when synthesized in Chlamydomonas • OPN is a highly acidic, phosphorylated protein • Triton has 14 colostrum OPN has very high concentration in human proteins in the pipeline • • Lead protein, OPN, has been colostrum (1.4 grams/liter), high concentration in successfully produced, human term milk (138 mg/liter), low concentration purified, validated for both in cow milk (9 mg/liter). immune protection and brain development biological • A large scale infant clinical trial activities has concluded OPN improves immune protection of infants • OPN has been shown to improve rat learning ability by regulating myelination
12 Triton has successfully produced hemoproteins in its yellow strain. This is a cost effective solution towards future clean meat
Impossibl Use Pichia (a Purify Add purified yeast) to make legehemoglbin legehemoglbin e Foods legehemoglbin from Pichia into burger
Use yellow Chlamy to make legehemoglbin Add whole algae Triton with hemoproteins Algae into clean meat Use yellow products Innovatio Chlamy to make beef or other ns myoglobin
13 Triton was elected as one of the top 30 “Cool Companies” by San Diego Venture Group in 2018
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