NON-FOOD APPLICATIONS

● Indigo and Dye

● Compost and Agriculture
● Energy Production
● Household Products
● Pharmaceuticals
 Natural Dyes Created via Fermentation

Cotton Denim
Fermented Indigo Your favorite blue jeans
INDIGOFERA TINCTORIA - Bacteria fermentation
to reduce indigo to a dye
--
Process Outline

1) Composted / dried indigo

2) Paste
3) Water
4) Alkalinity & PH
5) Organic material addition -
introduce natural LAB for
fermentation
6) Heat
7) Feeding the vat
• Complex microbial community "indigo-reducing bacteria" Alkalibacterium and Amphibacillus,
• Reduce the insoluble indigo molecule into a soluble form (leuco-indigo) under anaerobic and
alkaline conditions
Compost and Agriculture

“Turning human urine into a

resource”
● Developing agricultural
fertilizer and growth culture
“Bokashi” medium for microorganisms
via fermenting urine
Anaerobic putrefaction of compostables
• 40-75% Moisture, low O2
• Can also use high moisture like our cabbage ferments
Energy Production Biogases
Ethanol
● Ferment corn and sugarcane and harvest ethanol = biofuel
● Utilize enzyme to break down the carbohydrate into simple sugar
● Boil the wort
● Add yeast to ferment sugar into alcohol
● Distill the alcohol to make ethanol
● Concentrated ethanol gets distilled
● 200 proof (100% alcohol)
● Mix ethanol with gasoline to burn as fuel
Methane
● “Swamp gas”, “landfill gas”
● Product of anaerobic fermentation of sewage treatment and composting
● Capturing and refining biogas via a “digester”
● Turning pollutant into fuel
Hydrogen
● Fermentative pathways for producing biohydrogen
Biohydrogen via fermentative pathway
Pharmaceuticals
Medicinal applications
● Holistic eastern medicine utilizes
many fermented herbal elixirs
and alcohols or teas
Pharmaceuticals
Antibiotics and Other drugs
● Our most significant
antibacterial antibiotics
have been derived from
fungi
● Many molds and
fermented mushrooms

Alexander Flemming &

penicillin
Pharmaceuticals
Other Drugs
● Fermented tobacco --
developing flavor profile
through fermenting bulks of
tobacco leaves (like
compost) -- heat and
nitrogen by products
Insulin
● Making insulin from
recombinant DNA -
harnessing fermentation
Household Products

Skin care
● Japanese Cedar Enzyme Bath
○ Fermented pile of sawdust and rice bran
● Kombucha mother and kraut facials
● Fermented herbal skin care products
● Fermented ingredients in skin care products
Aromatherapy
● French Potpourri - “rotten pot”
Cleaning products
● Detergents and cleaning products rely on bio-enzymes
● Enzymes created and captured through fermentation
● Novozyme
Intro: Fermentation for
Alternative Proteins
 The world will have to close a 56% food 16

production gap by 2050.

Total calorie consumption globally
(trillion calories per
Crop production

56% food gap

year)

Note: Includes all crops intended for direct human consumption, animal feed, industrial uses, seeds, and biofuels. Source: WR I analysis based on FAO (2019a); UNDESA (2017); and
Alexandratos and Bruinsma (2012)
 Alternative Proteins and their production
How can we feed 10 billion people by 2050 and make the global food system better for the planet, people, and animals?

The world has a lot of people in it, and all those

people need to eat. People need to eat protein.
Plant Based Meat
Right now lots of people eat meat to get their
protein. We will not be able to grow enough
meat to feed the future population. We need
another, more sustainable way to produce Cultivated Meat
protein, this is the alternative protein industry.
Goal:
Cultivate meat and protein alternatives that Fermentation
have fewer trade offs in taste and texture,
that are commercially viable and tasty
without sacrificing on protein.
 18

Animal product alternatives will occur along a spectrum

Alternative Proteins and their production

Plant Based Meat

Cultivated Meat

Fermentation
 The general method used to produce plant-based
meat involves three primary steps. First, we grow
Alternative Proteins and their production crops as a source of raw materials. Second, we
process these crops to get rid of the parts of the
plants we don’t want. At this stage, we end up with the
Plant Based Meat proteins, fats, and fiber ingredients that will become
our plant-based meat product. Finally, we put
together the desired mixture of ingredients. This
ingredient mixture then goes through a manufacturing
process to create the muscle-like texture needed for
meat.

Base (tofu, Plant oils

Binding agent
tempeh, soy) (canola/sunflower)
(gluten/aquafaba/bean
s)
Alternative Proteins and their production

Cultivated Meat

Lab Grown Meat

Isha Datar, CEO of New Harvest, reveals why we need a post-animal bio-economy, driven by lab grown protein (meat, eggs, milk).

http://xprize.org

---------------------------------------
ABOUT XPRIZE

XPRIZE is an educational (501c3) nonprofit organization whose mission is to bring about radical breakthroughs for the benefit of humanity, thereby inspiring the
formation of new industries and the revitalization of markets that are currently stuck due to existing failures or a commonly held belief that a solution is not possible.
XPRIZE addresses the world's Grand Challenges by creating and managing large-scale, high-profile, incentivized prize competitions that stimulate investment in
research and development worth far more than the prize itself. It motivates and inspires brilliant innovators from all disciplines to leverage their intellectual and financial
capital.
 The manufacturing process begins with
acquiring and banking stem cells from an
animal. These cells are then grown in
Alternative Proteins and their production bioreactors (known colloquially as
cultivators) at high densities and volumes.
Similar to what happens inside an animal’s
Cultivated Meat body, the cells are fed an oxygen-rich cell
culture medium made up of basic nutrients
such as amino acids, glucose, vitamins, and
inorganic salts, and supplemented with
proteins and other growth factors.

Changes in the medium composition, often

in tandem with cues from a scaffolding
structure, trigger immature cells to
differentiate into the skeletal muscle, fat,
and connective tissues that make up meat.
The differentiated cells are then harvested,
prepared, and packaged into final products.
This process is expected to take between 2-
8 weeks, depending on what kind of meat is
being cultivated. Some companies are
pursuing a similar strategy to create milk
and other dairy products.
Alternative Proteins and their production

Plant Based Meat

Cultivated Meat

Fermentation
The Stir Tank Bioreactor/Fermenter
24

Impeller

Control software

Sensors to monitor pH and DO

Sparge

Vessel Temperature control

Sparge
Harvest
Alternative Proteins and their production

Fermentation ● Traditional
○ Modify plant-derived ingredients
● Biomass
○ Use fermentation to efficiently
produce proteins
○ Produce large quantities of protein
quickly
● Precision
○ Use microbial host as cell factory for
producing specific ingredients

GFI SYMPOSIUM ON
FERMENTATION
 Microbial Fermentation - Precision Fermentation and 26

Single Cell Protein

Pichia/Komagaetella yeast are budding
Submerged Fermentation (SmF) yeast that can be induced by methanol
to produce large amounts of secreted
protein. Used to ferment lehemoglobin
by Impossible Foods.

T. reesei yeast are filamentous yeast that

naturally secrete large amounts of
protein when starved. Used by Perfect
Day to produce whey protein.

Post-biotic bacteria are

Yarrowia yeast synthesize
anerobes that Superbrewed
and store fatty acids in their
uses for SCP.
cells.

Wikipedia
Airlift Fermentor – often for fungal cultivations

● Low energy input compared to stirred tank

fermentors.
● Less heat generated by friction
● Has potential for PF by filamentous/biofilm
fungi.
● Design/rate process optimization is non-trivial
● May not effectively distribute depending on
cell/nutrient mix.
 Segments of Alternative Protein Fermentation

Target Selection and Strain Feedstock Bioprocess

Design Development Optimization Design

End-product formulation and

manufacturing
Much of the Cost and Effort of Fermentation is 29

Downstream Processing
Seed
Host organism inoculation growth
Fermentor

Inoculate

Production Production run with protein

Fermentor secretion to media

Dewatering Harvest protein rich supernatant:

Clarification Centrifugation, TFF microfiltration

pH
adjustment pH adjustment to concentrate and
intermediate filtration (if needed)
and filtration

Concentratio
TFF ultrafiltration to concentrate
n/
and diafilter the protein
Diafiltration

Spray dry the concentrated protein

Drying
to a powder.
Alt protein investment backdrop
30

Source: GFI analysis of Pitchbook data

Fermentation-powered alternative protein 31

investments surpass $1 billion

Total invested capital: $3.48 billion

Source: GFI analysis of PitchBook Data, Inc. Invested capital includes accelerator and incubator funding, angel funding, seed funding, equity and product crowdfunding,
early-stage venture capital, late-stage venture capital, private equity growth/expansion, capitalization, corporate venture, joint venture, convertible debt, and general
debt completed deals.
Note: Data has not been reviewed by PitchBook analysts.
 32

Disruptions happen quickly

RethinkX report: Rethinking Food & Agriculture, 2020-2030

Key opportunities for innovation 33

Target identification and selection

Strain development

Feedstock optimization

Process design & manufacturing

Formulation & product development