Production of

Heterotrophic Microalgae
Presenter: Miller Tran, Ph.D.
Associate Director of Research and Development
Triton Algae Innovations
San Diego, California USA

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Microalgae are versatile group of organisms

Algae can perform photosynthesis to reduce CO2 or consume and organic

carbon source and undergo respiration to facilitate heterotrophic growth
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Definitions
• Heterotrophic: An organism that cannot manufacture its own
food and instead obtains its food and energy by taking in and
utilizing organic substances.
• Synonyms: heterotroph
• Heterotrophs include: all animals, protozoan, fungi, most
bacteria, some aquatic and marine algae, and some
cyanobacteria
• Heterotrophs: require nitrogen (N), phosphorus (P), other trace
elements and oxygen for their growth and productivity
• Aerobic Fermentation: Production of heterotrophic algae in the
presence of O2

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Differences between phototrophic and
heterotrophic growth processes

Heterotrophic
1 Growth in dark
2 Requires O2 to undergo respiration
3 Requires reduced carbon
4 High densities above 100g/L
5 Complete containment
Phototrophic
Growth in light
Requires CO2 to undergo
photosynthesis
No reduced carbon source necessary
Low densities below 10g/L
Contained or open reactors

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Examples of Different Organic Carbon Sources
• Monosaccharides: Glucose, fructose, & galactose
• Disaccharides: Sucrose, lactose, maltose
• Acetate
Monosaccharides Disaccharides Acetate

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How Heterotrophic Algae Metabolize Organic Carbon
Glucose Acetate

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Examples of Algae that can grow heterotrophically
Strain Microalgae Carbon Source Reference

1 Brachiomonas submarina Acetate Tsavalos & Day, 1994

2 Chlamydomonas reinhardtii Acetate Chen & Johns, 1996

3 Chlorella protothecoides (Auxenochlorella protothecoides) Glucose Chen & Walker 2012

4 Chlorella pyrenoidosa Acetate/glucose/lactose Running et al., 1994

5 Chlorella reguaris Acetate/glucose Endo et al. 1977

6 Chlorella saccharophila Glucose Tan & Johns, 1991

7 Chlorella sorokiniana Glucose Chen & Johns 1991

8 Chlorella vulgaris Acetate/glucose/lactose Glaude & Maxey, 1994

9 Crypthecodinium cohnii Glucose Barclay et al., 1994

10 Dunaliella salina Acetate/glucose/lactose Glaude & Maxey, 1994

11 Dunaliella tertiolecta Glucose Glaude & Maxey, 1994

12 Euglena gracilis Acetate/glucose Cook & Heinrich, 1965

13 Haematococcus pluvialis Acetate Kobayashi et al., 1997

14 Nannochloropsis oculata Glucose Glaude & Maxey, 1994

15 Nitzschia alba Glucose Barclay et al., 1994

16 Poterioochromonas malhamensis Glucose Kobayashi et al., 1997

17 Tetraselmis chuii Glucose Glaude & Maxey, 1994

18 Tetraselmis suecica Acetate/glucose/lactose Glaude & Maxey, 1994

19 Tetraselmis verrucosa Glucose Glaude & Maxey, 1994

20 Tertraselmis tetrathele Acetate/glucose/lactose Glaude & Maxey, 1994 7

Traits of Algae that grow well heterotrophically
• The ability to consume an organic carbon source
- In the absence of photosynthesis cells need reduced carbon to grow
• The ability to grow in light limited conditions
- High density aerobic fermentation is performed in stainless steel
production vessels that are devoid of light
• The ability to be sheer resistant
- Cultures are mixed with propellers going at high speeds to ensure
efficient gas exchange

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Examples of Systems to Produce Heterotrophic Algae

Research / Laboratory-Scale Pilot-Scale Systems

Triton: 500mL

Triton: 20L Triton Partner: 3,000L

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Examples of Systems to Produce Heterotrophic Algae
Commercial Scale Systems

Triton Partner: 100,000L Alltech: 100,000L

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Examples of Systems to Produce Heterotrophic Algae
Commercial Scale Systems

TerraVia & Bunge: 500,000L 11

Products Made From Heterotrophic Algae

DSM Omega-3 fatty acids from heterotrophic algae are added to a variety of products
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Products Made From Heterotrophic Algae

TerraVia cooking oil, butter, and flour are made from heterotrophic algae 13