Use of Microbial Consortia for Conversion of Biomass Pyrolysis - - PowerPoint PPT Presentation

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Use of Microbial Consortia for Conversion of Biomass Pyrolysis - - PowerPoint PPT Presentation

May 02, 2023 209 likes •451 views

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value- Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk Use of Microbial Consortia for Outline Conversion of Biomass Pyrolysis Liquids into Value-Added

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Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value- Added Products

Julian Pietrzyk julian.pietrzyk@ed.ac.uk

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Outline

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

  • Pyrolysis
  • Bio-oil
  • AD
  • Illumina Sequencing
  • Mass Spectrometry
  • Relatedness

Py-AD

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SLIDE 3
  • Biomass burned at high temperatures (300°C - 600°C) in the absence of oxygen
  • Thermal depolymerisation of lignocellulosic biomass
  • Products are char, bio-oil & syngas

Pyrolysis

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk Reactor Heat Trap 1 Heat Trap 2 Heat Trap 3 Cold Trap 1 Cold Trap 2 Receiver N2

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SLIDE 4
  • Product of pyrolysis of biomass
  • Dark brown organic liquid
  • High water content (~25 wt %)
  • Extremely high oxygen content
  • + 1000s other compounds
  • Ages instantly
  • Composition dependant
  • n feedstock
  • Low pH & biocatalyst inhibitors

catechol, guaiacol, syringol, isoeugenol, pyrones, vanillin, furans, acetic acid, formic acid, sugars, carboxylic acids, phenolics, hydroxyketones, hydroxyaldehydes

Bio-oil

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

Organic Aqueous

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Hydrolysis

  • High molecular weight organic polymers

split into smaller more bioavailable monomers.

  • Proteins > Amino acids | Carbohydrates >

Monosaccharides | Fats > Fatty acids | + H2

Acidogenesis

  • Acidogenic fermentation of hydrolysed

products to short chain…

  • volatile acids (propionic, butyric, acetic, formic,

lactic)

  • alcohols (ethanol, methanol)
  • H2 + CO2 + NH3 + H2S

Acetogenesis

  • Further digestion of acids by acetogens to

H2, CO2 and acetic acid.

Methanogenesis

  • Methanogenic archaea convert H2 and

acetic acid to CH4, CO2.

Anaerobic Digestion

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

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Vast range of microorganisms capable of bioconversion across a spectrum no single species could accomplish. An ideal platform for the detoxification of complex

  • rganic mixtures such as bio-
  • il.

Enables relevant primary energy savings of non- renewable sources without worsening abiotic resources depletion + a strong reduction of GHGs emissions.

(Fabbri & Torri, 2016)

Py-AD

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

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Hohenheim Biogas Yield Test

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

  • pening for

sampling stopcock gas compartment glass syringe substrate plunger scale silicone layer

  • 12 × 100 ml glass syringes
  • 30 ml Seafield water treatment plant anaerobic digestate
  • Supplemented with 10 g/l COD bio-oil, dried anaerobic digestate

(AD), wood pellets (WP) or seaweed (SW)

  • Mesophilic (~37°C) for 102 days

(adapted Mittweg et al., 2012)

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Biogas

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

50 100 150 200 250 10 20 30 40 50 60 70 80 90 100 Biogas Generated (ml) Time (days) DIGESTATE AD BO WP BO SW BO

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Illumina sequencing

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

Target gene 16S rRNA V4 hypervariable region Conserved region Conserved region 5’ Illumina adapter (adapted) Primer pad Primer linker Forward primer Read 1 Read 2 Index Reverse primer Primer linker Primer pad Golay barcode 3’ Illumina adapter

20 40 60 80 100 V3 V4 aV4 Coverage (%) Archaea Bacteria V4 aV4

Adapted V4 forward primer

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Illumina sequencing

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

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Illumina sequencing

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

10 20 30 40 50 60 70 80 90 100 d0 DIG1 d0 DIG2 d0 DIG3 d102 DIG1 d102 DIG2 d102 DIG3 d102 AD1 d102 AD2 d102 AD3 d102 MP1 d102 MP2 d102 MP3 d102 SWP1 d102 SWP2 d102 SWP3 Relative Abundance (%)

Phylum Class Order Family Genus Euryarchaeota Methanobacteria Methanobacteriales Methanobacteriaceae Methanobrevibacter Euryarchaeota Methanomicrobia Methanosarcinales Methanosaetaceae Methanosaeta Euryarchaeota Methanobacteria Methanobacteriales Methanobacteriaceae Methanobacterium Lokiarchaeota uncultured uncultured uncultured uncultured WSA2 WCHA1-57 uncultured uncultured uncultured Euryarchaeota Methanomicrobia Methanosarcinales Methanosarcinaceae Methanosarcina Lokiarchaeota uncultured uncultured uncultured uncultured Lokiarchaeota uncultured uncultured uncultured uncultured Euryarchaeota Methanomicrobia Methanomicrobiales Methanomicrobiaceae Methanoculleus Euryarchaeota Thermoplasmata Thermoplasmatales Marine Benthic Group D and DHVEG-1 uncultured

Archaeal top 10

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Illumina sequencing

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

10 20 30 40 50 60 70 80 90 100 d0 DIG1 d0 DIG2 d0 DIG3 d102 DIG1 d102 DIG2 d102 DIG3 d102 AD1 d102 AD2 d102 AD3 d102 MP1 d102 MP2 d102 MP3 d102 SWP1 d102 SWP2 d102 SWP3

Phylum Class Order Family Genus Thermotogae Thermotogae Petrotogales Petrotogaceae Defluviitoga Cloacimonetes W5 uncultured uncultured uncultured Bacteroidetes Sphingobacteriia Sphingobacteriales Lentimicrobiaceae uncultured Bacteroidetes Bacteroidia Bacteroidales Porphyromonadaceae Proteiniphilum Firmicutes Clostridia D8A-2 uncultured uncultured Firmicutes Clostridia Thermoanaerobacterales Thermoanaerobacteraceae Gelria Bacteroidetes Bacteroidia Bacteroidales Porphyromonadaceae uncultured Proteobacteria Gammaproteobacteria Pseudomonadales Pseudomonadaceae Pseudomonas Firmicutes Clostridia Clostridiales Caldicoprobacteraceae Caldicoprobacter Firmicutes BSA1B-03 uncultured uncultured uncultured

Relative Abundance (%)

Bacterial top 10

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Illumina sequencing

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk d0 DIG d102 DIG d102 AD d102 SW d102 WP

  • 1.2
  • 0.6

0.6 1.2

  • 2
  • 1

1 2 NMDS 2 NMDS 1 Archaea d0 DIG d102 DIG d102 AD d102 SW d102 WP

  • 1
  • 0.5

0.5 1

  • 2
  • 1.5
  • 1
  • 0.5

0.5 1 NMDS 2 NMDS 1 Bacteria

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ESI FT-ICR MS

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

ESI FT-ICR MS DCNC CJIℏ⊙Щ M CNIO RW∞

Van Krevelen diagram

1 2 0.0 0.5 1.0 H/C O/C carbohydrate lignin condensed hydrocarbon lipid protein 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 200 250 300 350 400 450 500 Intensity ×108 m/z 0.0 1.0 2.0 3.0 299.05 299.10 299.15 Intensity ×107 m/z

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ESI FT-ICR MS

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

d0 Digestate d102 Digestate d0 AD/BO d102 AD/BO d0 SW/BO d102 SW/BO d0 WP/BO d102 WP/BO

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ESI FT-ICR MS

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk d102 DIG1 d102 DIG2 d102 DIG3 d102 AD2 d102 AD1 d102 AD3 d102 SW1 d102 SW2 d102 SW3 d102 WP1 d102 WP2 d102 WP3

  • 1.5
  • 1
  • 0.5

0.5 1 1.5

  • 2
  • 1

1 2 NMDS 2 NMDS 1

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Relatedness

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

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Relatedness

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk d102 DIG1 d102 DIG2 d102 DIG3 d102 AD1 d102 AD2 d102 AD3 d102 SW1 d102 SW2 d102 SW3 d102 WP1 d102 WP2 d102 WP3

  • 30
  • 20
  • 10

10 20 30 40

  • 40
  • 30
  • 20
  • 10

10 20 30 40 Defluviitoga Abundance (24.4% of total variation) Cloacimonetes Abundance (38.2% of total variation)

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Relatedness

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

Analysis:

  • Distance-based linear model (DistLM).
  • Multivariate chemical data matrix using

the abundance profiles of Candidatus. Cloacimonetes and the Defluviitoga as the predictor variables.

  • The predictor variables are additionally

plotted as vectors (annotated arrows).

  • The abundance profiles of these two

microorganisms are cumulatively able to explain 62.61% of the chemical variation

  • bserved.
  • Candidatus Cloacimonetes phylum abundance correlates with the chemical

pattern separating reactor conditions – propionate degradation?

  • Defluviitoga suggests that increases in its abundance are related to the

chemistry observed for digestate-only control reactors – specific inhibition by bio-oil?

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Importance

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

  • Patterns in reactor chemistry can be correlated to

fluxes in microbial community

  • Understanding the microbial players involved at

each stage of AD

  • Longitudinal studies: continuous sampling of

both the chemical and biological species involved to identify process bottlenecks

  • Strategies to overcome inhibition
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What next?

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk

Scanning electron microscope image

  • f a biochar surface. From Schulz, H.

& Glaser, B. (2012).

Biochar

Biochar supplementation aids AD by the adsorption of inhibitory compounds and via the adherence of microbial cells in biofilms.

  • High surface area, biofilm formation
  • Biofilms show increased resistance to

environmental stresses

  • Partially conductive to the flow of electrons,

capable of supporting direct interspecies electron transfer (DIET)

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Thank you!

Use of Microbial Consortia for Conversion of Biomass Pyrolysis Liquids into Value-Added Products Julian Pietrzyk julian.pietrzyk@ed.ac.uk