Synthetic Biology and Industrial Biotechnology
Jens NielsenSystems and Synthetic Biology
ChalmersGothenburg, Sweden
Volume of Product
Valu
e of
Pro
duct
Pharmaceuticals
Fine Chemicals
Food Ingredients
Bulk Chemicals
Product value vs. volume
AntibioticsChiral building blocksEnzymes
VitaminsAntioxidantsFlavours
EthanolSolventsFeed additives (amino acids)Polymer building blocks
Over the whole range of products it is relevant to develop efficient cell factories for industrial production
Recombinant proteinsStatins and other natural products
Biotech Products
2
Chemical sales(100%=€1.383 billion)
Biotech sales(100%=€48 billion)
Between 2007 and 2012 chemical sales is expected to grow about 25% (to €1.748 billion) whereas biotech sales is expected to grow by more than 180% (to €135 billion)
Festel (2010) Ind. Biotech. 6:88-94
The Chemical Market
3
Enzymes
Polylactate
Sorona
Cephalexin
Vitamin B2
Move towards a biobased economy
Many success stories already implemented
Many novel processes are in the development phase
Time
White Biotechnology
Bio-feedstocks• Replacement of fossil fuel by sugars and starch
Bio-processes• Production of chemicalsusing bioprocesses, e.g. vitamins
Bio-products• polymers• enzymes• food ingredients
Today
Marked
Thepotential
Industrial Biotechnology
4
Pre‐treatment
Fermen‐tation
Purifi‐cation
Formu‐lation
RawMaterial
Chemicals&
Fuels
The research intensive partDesire to develop plug and play
solutions, i.e. common cell factory platforms for many
products
The Value Chain in Industrial Biotechnology
Biobutanol
2. Generationbioethanol
There is a trend towards partnerships – sharing competences, risks, and capital investments5
Yeast is a widely used cell factoryEthanolBaker’s yeastWine & BeerResveratrolInsulin precursorsVaccines (HPV, Hepatatis)
Many ongoing attempts to develop novel cell factoriesProduction of fuels (butanol, biodiesel), commodity chemicals (malate, succinate, 3-OH propionic acid), fine chemicals (isoprenoids), food ingredients (PUFAs) and protein drugs
Platform Cell Factories
6
Metabolic Engineering
The enabling technology that will allow for design of novel cell factories that can be constructed through the use of genetic engineering
The genomics revolution has resulted in a technology pushMetabolic Engineering:• bioanalytics• genomics• quantitative physiology
Xylitol
Phosphate Glu NAD
Xylitol
Phosphate Glu NADPhosphate Glu NAD
Enabling Technologies
7
Industrial Systems Biology
B&B (2010) 105:439-4608
+
+
Components and their abundance
Interactions and information flow
SystemMecha
nical
system
Biological
system
Systems Biology
9FEMS Yeast Research (2008) 8:122-131
Ana Paula Oliveira, PhD thesis (2008) DTU
Can we expand in silico design and analysis of cell factories?
Yeast Platform for Succinate ProductionModel guided ME strategy
SerineGlycine Biomass Demand Succinate Biomass
Coupled
Reference (CEN.PK113‐5D)
Mutant 8D(Δsdh3 Δser3Δser33)
+ 500 mg/L glycine
Genetic engineering
Specific Growth Rate (1/h)
Maximum Titer (g/L)
Maximum Yield (g/g‐biomass)
Maximum Yield (g/g‐glucose)
REF 0.33 0.03 0.01 0.008D 0.22 0.40 0.14 0.02
0
0.1
0.2
0.3
0.4
0.5
Proof of Concept
10
Evolved 8D(Δsdh3 Δser3 Δser33)
Mutant 8D(Δsdh3 Δser3Δser33) 500 375 250 125 25
+ mg/L glycine supplemented, Directed Evolution
0
Select for Faster Growth
0.09 0.100.13
0.10
0.39
0.69
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
YXSuc[g‐suc/g‐DCW]
500 mg/L375 mg/L250 mg/L125 mg/L25 mg/L0 mg/L
Glycine Supplemented
SF1 SF2 SF3
Growth Rate 0.03 0.08 0.14Yield 0.69 0.27 0.27
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
YXSuc[g‐suc/g‐DCW]
Growth Rate [1/h]
Growth Rate
Yield
Error bars represent ± 99% CI (n=3)
Yeast Platform for Succinate ProductionThrough evolution a glycine-prototroph could be evolved
11
Substrate A B
C D E
Product
By-product
X Y
Substrate A B
C D E
Product
By-product
X Y
Substrate A B ProductX Y
Metabolic Engineering
Synthetic Biology
Metabolic Engineering vs.Synthetic Biology
12
Glucose
Sucrose
Xylose
Arabinose
Fine chemicals
Biofuels
Commodity chemicals
Protein drugs
Volume
Value
Butanol
Biodiesel
3-HPA
Succinic acid
Sesquiterpenes
Antibiotics
Human insulin
S. cerevisiae
Yeast as a Platform OrganismAn extensive technology base for wider use of yeast as platform cell factory has been established in our group
Metabolic Engineering
Systems Biology
13
Acknowledgements Funding
YSBN
14