Download White Dog Labs Introduction

Maximum Yield and
Zero CO2 MixoFerm™
Shawn Jones
BIO World Congress 2016
“Those brave pioneers distilled whiskey and drank it right off the still.
They called it White Dog”
Confidential and Proprietary Materials of White Dog Labs, Inc
White Dog Labs
Micro &
Molecular Biology
Fermentation & Separation
Process Development
R12
Plant modeling
and design
Confidential and Proprietary Materials of White Dog Labs, Inc2
Biomass to Chemical/Fuel
Conundrum
FEEDSTOCK
Average degree of
reduction = 4 - 4.5
Currently:
Oxidize some biomass to CO2
and sacrifice mass yield
Our approach:
Provide some reductant
(H2 and/or H2 & CO) and
maximize mass yield
PRODUCTS
Average degree of
reduction = 5 - 6
More reduced than feedstock.
So requires ENERGY!
Confidential and Proprietary Materials of White Dog Labs, Inc
WDL’s MixoFerm™ Technology
Gaseous Feedstocks
Municipal
Solids
Waste
Biomass
Coal
Carbohydrate Feedstocks
Natural
gas
Gas and/or Methanol
1st
Gen Sugars
2nd
Gen Sugars
Glycerin
(biodiesel
waste)
Acid whey
Carbohydrate
Mixotrophic Fermentation
(i.e., simultaneous fermentation of gas with sugar)
Maximizes sugar yield, minimizes CO2 emissions
and synergizes fossil with bio.
R12
Confidential and Proprietary Materials of White Dog Labs, Inc4
MixoFerm™ at its core – at the
cellular level
CO2
H2
CO
CO2
Glycolysis
Pathway
Optional
Addition
Wood-Ljungdahl
Pathway
Biochemicals:
Acetone
Isopropanol
N-Butanol
2-Pentanol
N-Hexanol
Ethanol
Alkanes
Alkenes
Fatty Alcohols
Cell Environment
Acetyl-CoA
Cell Interior
Glucose
The novel WDL “mixotrophy” process is a method of
incorporating two separate pathways to produce
multiple biobased chemicals
Reducing gases can be renewable,
non-CO2 emitting and/or mitigating waste:
• H2 from electrolysis of water – solar,
wind or hydroelectricity input
• Biomass gasification
• Steel mill effluent gases
• Other waste gases
• Syngas from biogas
Confidential and Proprietary Materials of White Dog Labs, Inc
WDL’s MixoFerm™ Target Molecules
Mixotrophy
maximizes acetylCoA formation.
Acetyl-CoA is
building block of
nearly all
biochemicals.
R12
Confidential and Proprietary Materials of White Dog Labs, Inc6
Modes of MixoFerm™
Sugar
Traditional
Fermentation
Metabolite
(product)
Sugar
Sugar
CO2 H2
MixoFerm™
(no gas addition)
Metabolite
(product)
CO2
H2
MixoFerm Plus™
(with H2 addition)
Metabolite
(product)
Sugar
CO/H2
MixoFerm Plus™
(with syngas
addition)
Metabolite
(product)
Mixotrophy can be implemented through multiple routes
Confidential and Proprietary Materials of White Dog Labs, Inc
Potential Product Advantages through
MixoFerm™
20-65 kg H2/ton of product needs to be added
Mass yields can
be improved
50 – 100% for
essentially ANY
biochemical
Maximum Yield (g/g glucose)
0.8
Traditional
MixoFerm
MixoFerm Plus
0.6
0.4
0.2
0.0
R12
Confidential and Proprietary Materials of White Dog Labs, Inc8
Demonstration of MixoFerm Plus™
80%
8.0
60%
6.0
40%
4.0
20%
2.0
0.0
0%
0
24
48
72
96
120
144
168
Time (hr)
Acetate (%)
R12
% Acetate Labeled (13C)
Strain utilized gaseous
feedstock in the
presence of excess sugar
100%
10.0
Concentration (g/l)
C. ljungdahlii grown on:
• 12C-sugar substrate
• 13C-syngas substrate
12.0
Fructose
Acetate
Confidential and Proprietary Materials of White Dog Labs, Inc9
Broad host range for MixoFerm™
Yield (CM/CS)
Mixotrophy displays
broad host range over
acetogens, though
some perform better
than others
R12
CAC
CLJ CAU MTA ELM
CLJ CAU MTA ELM
Traditional
fermentation
MixoFerm™
MixoFerm Plus™
10
Confidential and Proprietary Materials of White Dog Labs, Inc
Demonstration with Acetone
Strain #1:
Expression of acetone pathway in Clj
101wt% yield total
Need to reduce side
products
Strain #2:
Deletion of SADH gene
Expression of acetone pathway in Clj
80wt% yield total
Need to increase
yield of acetone
Strain #3:
Deletion of SADH gene
Expression of optimized acetone pathway in Clj
55wt% yield total
Achieved >90% of
theoretical yield
Able to develop and demonstrate for non-native metabolites
R12
11
Confidential and Proprietary Materials of White Dog Labs, Inc
Continuous fermentation for acetone
Demonstrated:
• Yield: 45wt%
• Productivity: 5 g/L/hr
R12
12
Confidential and Proprietary Materials of White Dog Labs, Inc
Demonstration of MixoFerm Plus™
1.0
Carbon Fraction
Clj acetone strain grown on:
• MixoFerm™
sugar with N2 headspace
• MixoFerm Plus™
sugar with 20%v/v H2 headspace
Theoretical maximum
from traditional
fermentation
0.8
0.6
CO2
Metabolites
0.4
0.2
MixoFerm Plus™ fermentation
emitted zero CO2
R12
0.0
MixoFerm
MixoFerm Plus
13
Confidential and Proprietary Materials of White Dog Labs, Inc
Potential for MixoFerm™
•
Implementation of MixoFerm™ or MixoFerm Plus™
maximizes biogenic carbon conversion to product
•
Biogenic carbon is the most expensive input into the
process, even compared to gas supplements
•
Broad applicability both of hosts and of products that can be
produced (anything from acetyl-CoA)
Confidential and Proprietary Materials of White Dog Labs, Inc