culture collection yerseke (ccy) -...
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Culture Collection Yerseke (CCY)
A collection of microalgae and cyanobacteria, for strain
selection, growth optimization, and product maximization
6/5/2015
www.intesusal.eu 1
Lucas J. Stal
Department of Marine Microbiology
Royal Netherlands Institute for Sea Research
Professor of Marine Microbiology
University of Amsterdam
6/5/2015
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Mission Statement
The Culture Collection Yerseke (CCY) is a non-profit facility that is specialized
in the cultivation of (marine) phototrophic microorganisms. The collection
comprises unicellular algae (green algae and diatoms) and cyanobacteria. The
collection now also extends to anoxygenic phototrophs and other bacteria that
live in marine phototrophic communities such as microbial mats and biofilms or
planktonic assemblages. CCY was established in 1997 by the Department of
Marine Microbiology of the Netherlands Institute of Ecology (NIOO-KNAW),
now the Royal Netherlands Institute for Sea Research (NIOZ), in Yerseke.
CCY provides a platform for maintaining novel strains of microorganisms that
have been isolated in the course of scientific research projects. These strains
are maintained alive as cultures or cryo-preserved. CCY provides expertise on
isolation, cultivation, and identification of novel strains and on their cryo-
preservation. The collection is available to academic and commercial parties,
except when agreements about specific strains prevent this.
CCY Collection Media How to Order Projects News Contact
CCY.NIOZ.NL
Mission Statement
The Culture Collection Yerseke (CCY) is a non-profit facility that is specialized in
the cultivation of (marine) phototrophic microorganisms. The collection
comprises unicellular algae (green algae and diatoms) and cyanobacteria. The
collection now also extends to anoxygenic phototrophs and other bacteria that
live in marine phototrophic communities such as microbial mats and biofilms or
planktonic assemblages. CCY was established in 1997 by the Department of
Marine Microbiology of the Netherlands Institute of Ecology (NIOO-KNAW), now
the Royal Netherlands Institute for Sea Research (NIOZ), in Yerseke. CCY
provides a platform for maintaining novel strains of microorganisms that have
been isolated in the course of scientific research projects. These strains are
maintained alive as cultures or cryo-preserved. CCY provides expertise on
isolation, cultivation, and identification of novel strains and on their cryo-
preservation. The collection is available to academic and commercial parties,
except when agreements about specific strains prevent this.
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Culture collections are not stamp or coin
collections
Why collect cultures?
Cultures are genetic repositories
Isolation, purification and cultivation
Characterization and identification
Optimization of growth media and cultivation conditions
Screening for bioactives and other valuable compounds
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Collection of ~700 microorganisms:
mainly photoautotrophs
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Prorocentrum lima Green algae
Tetraselmis striata Phaeodactylum tricornutum
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CO2
O2
Fatty acid,
sugars,
proteins..
Pediastrum CCY0386
Photoautotrophs
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Cryopreservation
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Cryopreservation
Necessary because:
• Guarantee the stability of the genetic composition
• To avoid human errors (e.g. mislabeling and contamination)
• Decrease maintenance costs and labor-intensive activities (e.g. sub-
culturing, medium preparation, glassware cleaning)
• Allows the collection of many more cultures
• Needs minimum storage space
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Controlling the rates of cooling:
• Controlled-cooling canister as Mr. Frosty (approx. -1C per min -1)
• Controlled Rate Freezer (-XC per min -1)
Mr. Frosty
Isopropanol resevoir
Cryovial holder
Cryomed1010
Controlled rate freezer
Cryopreservation
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plasmolysis Intracellular ice
Cryopreservation: cooling rate
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CryoProtective Agents (CPAs):
• Penetrating Agents (MeOH,
DMSO, Glycerol)
• Lower the intracellular water
freezing-point
• Decrease plasmolysis
Cryopreservation
Phormidium
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Lipid droplets in Phaeodactylum
stained with Bodipy. The chloroplast
fluoresces red.
Lipid droplets in Phaeodactylum
stained with Nile Red
Lipids
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408 strains of the Culture Collection Yerseke (CCY) were tested
The BIODIPY test revealed 220 positive strains (in which green
fluorescence was detected), of which 165 showed strong
fluorescence or many fluorescing lipid bodies.
CCY0216 (Nitzschia sp., diatom) CCY0224 (Surirella brebisonii, diatom) CCY0305 (Odontella sp., diatom)
CCY0399 (Navicula sp., diatom) CCY0348 (Phormidium sp., cyanobacteria) CCY9611 (Gloeothece sp. cyanobacteria)
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100% N 50% N 10% N 0% N
BODIPY labeling of CCY0033 after nitrogen starvation
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CCY0033
CCY0438
CCY0913
+glycerol
+glycerol
+glycerol
Lipids droplets in Phaeodactylum CCY0033
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The cells are almost completely filled with two lipid droplets
Effect of glycerol on lipid accumulation
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6/5/2015 InteSusAl - confidential 20
Culture fed with glycerol (50 g/l) Culture did not receive glycerol
Effect of glycerol on cell size
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Growth condition Cell size (μm)
14⁰C-100%N 4.7
14⁰C, 100%N + 50g/l glycerol 5.3
23⁰C, 100%N 4.4
23⁰C, 100%N + 50 g/l glycerol 5.2
23⁰C, 10%N 4.4
23⁰C, 10%N + 50g/l glycerol 5.4
Cells 13% (14oC), 18% (23oC, 100% N) or 23% (23oC, 10% N) bigger when fed with glycerol
Lipid content (% of dry weight)
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Growth condition % lipids C:N Pigments
pg/cell
POC
mmol/l
PON
mmol/l
14⁰C, 100%N 7.1 9 0.51 23.9 2.68
14⁰C, 100%N + 50g/l glycerol 28.8 25 0.36 72.1 2.87
23⁰C, 100%N 11.4 8 0.63 15.3 2.03
23⁰C, 100%N + 50 g/l glycerol 33.0 23 0.42 79.5 3.43
23⁰C, 10%N 12.2 13 0.45 18.1 1.44
23⁰C, 10%N + 50g/l glycerol 47.5 78 0.15 66.5 0.86
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The Marine Microbiome Applied (MaMA)
‘Where Industry meets Academia’
An Industry Conference back-to-back to an Open Science
Meeting on the biotechnical application of marine microbes
organized by MaCuMBA
Berlin-Adlershof
27-30 June 2016
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www.macumbaproject.eu
ANNOUNCING: