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Adaptation to environmental conditions Martin Könneke www.icbm.de

Adaptation to environmental conditionsTemperature adaptation of bacterial membrane fatty acids saturated fatty acids long chain fatty acids Desaturase De novo- synthesis De novo- synthesis

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Adaptation to environmental conditions

Martin Könneke www.icbm.de

Adaptation

•! What environmental factors affect growth?

•! Cell structure and growth

•! Adaptations to environmental factors •! Cold adaptation in SRB •! Growth above 100 ˚C

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Pressure •! Light and radiation •! (Energy & carbon source)

Extreme <-> Normal

Specialist <-> Generalist

Growth <-> Survival

Permanent <-> Transient

Obligate <-> Tolerant

Growth

Transport

Biosynthesis Maintenance of highly organized structure Regulation

Reproduction Detoxification

Macromolecules in the cell

Quelle: Brock Biology of Microorganisms

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Pressure

Quelle: Brock Biology of Microorganisms

Temperature limits for life

•! Freezing point in seawater around -2 ˚C

•! Partially liquid in antarctic sea ice at -15 ˚C •! Liquid up to 300 ˚C at high pressure •! Only Prokaryotes grow above 65 ˚C

•! Upper limit: strain 121 grows at 121 ˚C

Upper limit: strain 121 grows at 121 ˚C

Kashefi and Lovley 2003

The acido- and thermophilic algae Cyanidium caldarium

Yellowstone NP, (~50 ˚C, pH 2)

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Pressure

Quelle: Brock Biology of Microorganisms

pH and microbial growth

•! pH range for growth limited to 2 units

•! How can they generate proton motive force? •! Picrophilus oshimae (pH 0.7 - 2)

•! Sea water in general well buffered around pH 8

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Energy & carbon source

Water activity, osmosis, and halophiles

•! All organisms require water •! Water content and substances dissolved in water •! Sea water contains about 3% NaCl •! Halophiles: salt-loving organisms

Quelle: Brock Biology of Microorganisms

Quelle: Brock Biology of Microorganisms

Water activity, osmosis, and halophiles

•! All organisms require water •! Water content and substances dissolved in water •! Sea water contains about 3% NaCl •! Halophiles •! Osmophiles: live in environments high in sugars •! Xerophiles: live in very dry environemnts

Compatible solutes allow growth under conditions of low water activity.

Quelle: Brock Biology of Microorganisms

Compatible solutes increase the internal solute concentration.

Quelle: Brock Biology of Microorganisms

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Pressure

Toxic forms of oxygen (By-products during O2 reduction)

Quelle: Brock Biology of Microorganisms

What environmental factors affect growth?

•! Temperature

•! pH •! Water availability •! Oxygen

•! Pressure

Quelle: Brock Biology of Microorganisms

Does pressure affect microbial growth?

Pressure increase by 1 bar per 10 m water depth

How can they stand 100 bar in 1000m depth?

Bert Engelen

Temperature adaptation in SRB

Dissimilatory sulfate-reduction occurs between -1.8 ˚C (arctic sediment) and above 100 ˚C (Guaymas basin sediment)

High phylogenetic and physiological diversity

permantly < 5°C

SO42- H2S

SO42- H2S

SO42- H2S

SO42- SO4

2-

changing temperatures

Tropical and

moderate climate

Polar regions

More than 90 % of the ocean biosphere is permantly cold!

Cold adaptations of microorganisms

•! Effects on growth rate and growth yield

•! Metabolic activity •! Protein flexibility and cold-adapted

enzymes

•! Protein synthesis and cold-induced proteins •! Effects on membrane and lipid structure

Isfjorden (Station D)

Kongsfjorden (Station F)

Bremen

Smeerenburgfjorden (Station J)

Krossfjorden (Station G)

Ny Ålesund

Longyearbyen D

G

J

Spitzbergen

0 °C

10 °C 20 °C 30 °C 40 °C

Ice

Mesophilic Most isolated SRB (since Beijerink, 1895)

Psychrotolerant: e.g. Desulfobacter hydrogenophilus (Widdel, 1987)

Psychrophilic Isolates from Arctic sediments (Knoblauch et al., 1999)

Temperature range

Optimal temperature

Fluid-mosaic structure of the membrane

Quelle: Brock Biology of Microorganisms

saturated fatty acids

cis-unsaturated fatty acids

short-chain fatty acids

branched fatty acids

Low temperature High temperature

Temperature adaptation of bacterial membrane fatty acids

saturated fatty acids

long chain fatty acids

Desaturase De novo- synthesis

De novo- synthesis

De novo- synthesis

4 °C 12 °C 20 °C 28 °C

Desulfofaba gelida

Desulfofrigus oceanense

Desulfotalea psychrophila

Desulfotalea arctica

40%

73%

82%

85%

70%

78%

83%

contains high amounts(>70%) of short chain fatty acids

Effect of temperature on unsaturated fatty acids in psychrophilic SRB

Desulforhopalus vacuolatus

85% 81%

Total amount of unsaturated fatty acids

Desulfovibrio desulfuricans

Desulfococcus multivorans

Desulfosarcina variabilis

49%

25%

23%

23%

22%

Desulfobacter hydrogenophilus

65% 27% 43% 61%

50%

Desulfobacter postgatei

43% 11% 24%

4 °C 12 °C 20 °C 28 °C

Effect of temperature on unsaturated fatty acids in mesophilic SRB

Total amount of unsaturated fatty acids

0

10

20

30

40

50

0 5 10 15

cyc 17:0

c9 16:1

10Me 16:0

16:0

0,00

0,10

0,20

0,30

0,40

0 5 10 15

Time (days)

0,4

0,3

0,2

0,1

OD

660 n

m"

Fatty a

cid

conte

nt (%

)"

0"

0,1"

0,2"

0,3"

0,4"

0" 10" 20" 30" 40" 50

Time (days)

OD

66

0 n

m"

0"

10"

20"

30"

40"

50"

0 200 400 600 800 1000

Fatty a

cid

conte

nt (%

)"

16:0

c9 16:1

10Me 16:0

cyc 17:0

12 °C 28 °C Precultured at 28 °C

Time course of changes in cellualr fatty acids at different growth-temperatures in D. hydrogenophilus

Changes in the fatty acid composition of Desulfobacter hydrogenophilus

Saturated Fatty acids

16:0

Cyclopropane Fatty acids

cyc17:0

cis-unsaturated Fatty acids

c9 16:1

De novo synthesis

CFA-Synthase

Low Temperature High Temperature

(Stationary growth-phase)

Protein pattern of Desulfobacterium autotrophicum Grown at different temperatures

•! exhibited constant fatty acid composition with high amounts of unsaturated or short-chain fatty acids.

•! are “specialists“ in a small temperature range, in which they probably outcomplete mesophilic/ psychrotolerant species.

•! change the ratio of saturated, unsaturated and cyclopropane containg fatty acids with changing temperature via de novo synthesis.

•! are “generalists“, which propably outcomplete psychrophiles in environments with changing tempeartures.

Psychrophiles (permantly cold sediments)

Psychrotolerants (moderate climate)

Quelle: Brock Biology of Microorganisms

Black Smoker (hydrothermal vent)

C. Cary

So-called "black smokers" are the hottest of the vents. They spew mostly iron and sulfide, which combine to form iron monosulfide. This compound gives the smoker its black color.

Endospores are differentiated cells that very resistant to heat and harsh chemicals.

Quelle: Brock Biology of Microorganisms

Lipid bonds in Bacteria, Eukarya and Archaea

Quelle: Brock Biology of Microorganisms

Etherlipids in Archaea

Side chains consist of repeating isoprene units!

Quelle: Brock Biology of Microorganisms

Structure of archaeal membranes

Glycerol-diether Glycerol-tetraether

Quelle: Brock Biology of Microorganisms

In general,temperature is the most important environmental factor in marine systems.

…and easy to reproduce in the lab!