SBS-713 Archaea, the third domain of life

Chitin and Chitin metabolism in Archaea

T. kodakarensis

Dr. Mehwish Aslam

Biorefinery based on renewable biomass
1) To avoid global warming and establish a sustainable
society, alternative resources are required to both
supplement and replace fossil fuels
2) Biomass is a promising candidate, as it is renewable
and prevalent over the Earth. Annual global biomass
production is ~220 billion dry tons or 45 x 1020 J, that is
equivalent to 8.3 times the world’s annual energy
consumption (5.43 x 1020 J, in 2014)
3) Biomass conversion into useful chemicals, biofuels
and materials is called “biorefinery processes”
expected to greatly expand in future economies
Chitin – an abundant, unused biomass

Chitin is a biomass of β-1,4-linked homopolymer of N-acetyl-

glucosamine (GlcNAc)
The 2nd most abundant natural polysaccharide after cellulose
(1010-1011 metric tons produced per year)
Structure of chitin

Chitin is mainly found in marine organisms such as shrimps,

crabs and cuttlefish. Other sources include bacteria,
arthropods, protozoans and fungi
Chitin biomass remains largely unused at present
Microbial chitinolytic pathways

GlcNAc : N-Acetylglucosamine GlcN6P : Glucosamine 6-phosphate

GlcN : Glucosamine Fru6P : Fructose 6-phosphate
GlcNAc6P : N-Acetylglucosamine 6-phosphate
Microbial conversion of biomass at high temperatures

Biomass degradation at high temperatures exhibits

Higher reaction rates (Higher efficiency)
Lower risk of contamination (Higher stability)

This will be possible by employing cells and/or

enzymes from (hyper)thermophiles

Hyperthermophiles
Organisms that show an optimum growth temperature of
over 80˚C
Found at hot springs, deep sea hydrothermal vents or
volcano areas
A hyperthermophilic archaeon, Thermococcus kodakarensis

Optimum growth temperature: 85˚C

Genome analysis completed
Gene manipulation techniques established 2,088,737 bp
2,306 ORFs

High potential to produce molecular hydrogen (H2)

One of few hyperthermophiles with a gene cluster for chitin
degradation
Gene cluster involved in chitin metabolism

GlmA GlmD ABC transporter Dac ChiA 1000 bp

TK1754 TK1765
Construction of T. kodakarensis KC01

No growth on chitin
 Catabolic pathway for chitin conversion in T. kodakarensis
Chitinolytic pathway
GlcNAc : N-Acetylglucosamine
Chitin GlcN : Glucosamine
ChiA GlcN6P : Glucosamine 6-phosphate
Fru6P : Fructose 6-phosphate
(GlcNAc)2 GAP : Glyceraldehyde 3-phosphate
AcOH GAPOR : GAP:ferredoxin oxidoreductase
Dac
POR : Pyruvate:ferredoxin oxidoreductase
Mbh : Membrane-bound [Ni-Fe]-hydrogenase
GlcN-GlcNAc ACS : Acetyl-CoA synthetase
GlmA

AcOH
H2-generation pathway
GlcNAc GlcN
Dac ADP
GlcNK 2H+ H2 x 2 2H+ H2 x2
AMP

GlcN6P Mbh Mbh

GlmD
NH3 Fdox Fdred Fdox Fdred
ADP + Pi ATP + CoA

Acetate
Glucose Fru6P GAP Pyruvate Acetyl-CoA (AcOH)
x2 +
GAPOR POR ACS
Glycolytic pathway CoA
CO2

Theoretical conversion from GlcNAc

C8H15NO6 (GlcNAc) + 4H2O = 3CH3COOH + NH3 + 2CO2 + 4H2