Cyanobacteria

Blue-green algae
Microalgae
Unicellular Algae of the plant kingdom
Cyanobacteria (blue-green algae)
Prokaryotes
Oxygenic photosynthesis
On earth for at least 2.8 x 10
9
years
Architects of our atmosphere
Many fix N
2

Important primary producer
Wide variety of metabolic capacities
Wide variety of ecological environments
Cyanobacteria
The oldest known fossils, dated 2.8 - 3.5 billion years old
Source unknown
Cyanobacterial Diversity
Cyanobacterial Mass Culture
http://operationalgae.com/Bioreactors.html
http://www.lowtechmagazine.com/2008/04/algae-fuel-biof.html
Some Cyanobacteria
Fix nitrogen from the air
Produce hydrogen gas
Regulate flotation
Grow as mats
Make secondary products
Are naturally transformable
Approaches
Extract naturally occurring molecules
Sugars for fermentation
Lipids for biodiesel
Some natural hydrocarbons
Metabolic engineering to improve strains
Designer organisms
Make and secrete pure products
e.g., jet fuel!


Ducat D. C., J. C. Way and P. A. Silver. 2011. Engineering cyanobacteria to generate high-value products. Trends Biotechnol. 29:95-103.
Engineered biochemical pathways in model cyanobacteria
Anabaena variabilis ATCC 29413, Anabaena sp. PCC7120, Synechocystis
PCC6803, Synechococus sp. PCC7002 & Synechococcus elongatus PCC7942
Needed Skills & Technology
Microbial ecology
Identify good strains
Genetics, molecular biology
Improve strains
Aquaculture
Grow in large scale
Engineering
Harvest, extract, refine products
Two-pronged Approach
Use model organisms to
identify novel genes
Modify pathways

Identify candidate
production strains
Develop genetic tools
Engineer production strains
Modify traits
Import pathways
Genetic Tools for Model Strains

Unicellular and filamentous
models
Transformation, conjugation
Regulated promoters
Libraries
Gene knockout
Gene over-expression
Reporters
Fluorescent, bioluminescent,
chromogenic






Synechococcus elongatus PCC 7942



Unicellular cyanobacterium, obligate photoautotroph
2.7 Mb genome (approx 2700 genes)
Naturally transformable
Homologous recombination for gene replacement
Strains Engineered to Secrete Fatty Acids
F
A

c
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c
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(
m
M
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Time (days)
Two-pronged Approach
Use model organisms to
identify novel genes
Modify pathways

Identify candidate production
strains
Develop genetic tools
Engineer production strains
Modify traits
Import pathways
Finding Production Strains
Collect sample Isolate strains
Identify and characterize
Scale up & harvest Grow outdoors Choose & precondition
0%
20%
40%
60%
80%
100%
8
4
9
1
9
2
9
9
1
0
7
1
1
3
1
1
6
1
1
8
1
2
7
C18:3
C18:2
C18:1
C18:0
C16:2
Cultivation in 1 acre pond
BL0902 grown at production scale
Heterologous expression of GFP from ashuttle plasmid
Two-pronged Approach
Use model organisms to
identify novel genes
Modify pathways

Identify candidate
production strains
Develop genetic tools
Engineer production strains
Modify traits
Import pathways