A promising new method to deliver
hydrogen to bioreactors
Better hydrogen supply to methanogens
4 H2O H2 O2 Biogas (CH4 + CO2) manure biomass digestate digester
Principle of BioP2G
Diffuse rubber tubing:
best way for the supply of hydrogen
Fisherbrand® 4,0 x 1,0 mm
ml H2 / minute MF-controler H2-FLUX H2-flow AMPTSII Pressure,
Two reactor setups: in situ and ex situ
bioreactor
bioreactor 22nd reactor (ex situ)
nd
reactor (ex situ)
bioreactor (in situ)
bioreactor (in situ)
biomass biomass biogas biogas H2 H2 ex situ: in situ: biogas
control
in situ
H2 biogas CH4/CO2 CO2 + 4H2 → CH4 + 2 H2O + E biomass biomass biogas CH4/CO2sugar beet pulp / rabbit feed mix sugar beet pulp / rabbit feed mix in situ H2 CO2 CH4 gaschromatograph
∞
∞
H2 biogas CH4/CO2 bioreactor ex situ biomass biogas CH4/CO2
sugar beet pulp / rabbit feed mix ex situ H2 CO2 CH4 gaschromatograph CO2 + 4H2 → CH4 + 2 H2O + E
control
H2
in situ Labscale 10 liter reactor set-up: in situ
CH4 Fix-CO2
biomass biomass
AMPTSII
Relative vol.%-CH
4*: control
* H2, CH4 and CO2 = 100 vol.%
1 8 15 22 29 36 43 R e la tiv e v ol .% -C H 4 days
Relative vol.%-CH
4*: in situ
* H2, CH4 and CO2 = 100 vol.%
1 8 15 22 29 36 43
1 ml*min-1 2 ml*min-1 3 ml*min-1 4 ml*min-1 5 ml*min-1 No hydrogen No hydrogen days R el at iv e vo l.% -C H 4
CH4*- production: Control vs in situ
}
322 Nml CH4/gr biomass 445 Nml CH4/gr biomass
* CO2 is captured in 3M NaOH
days N m l C H 4
• Adding hydrogen in situ to an existing biogas
plant results in more methane per unit
biomass
• Use of silicone rubber tubing is attractive
H2 FischerbrandISO3302 E1 8.5 meter; 4.0x 1.0 mm LOW-ΔP-FLOW bioreactor ex situ CH4 Fix-CO2 biomass
Labscale 10 liter reactor set-up: ex situ
AMPTSII
Relative vol.%-CH
4*: bioreactor
* H2, CH4 and CO2 = 100 vol.%
R e la tiv e vo l.% -C H 4 days 1 8 15 22 29 36
Relative vol.%-CH
4*: ex situ
* H2, CH4 and CO2 = 100 vol.%
1 ml*min-1 2 ml*min -1 3 ml*min-1 4 ml*min-1 No hydrogen 1 8 15 22 29 36 R el a tiv e vo l.% -C H 4 No hydrogen days
Bioreactor LOW-ΔP-flow bioP2GasUnit
CH4+CO2 H2 + CH4 theoretical CH4 experimental CH4 experimental VS CH4 theoretical CH4 experimental VS CH4 theoretical Δ % = -7,2Issue:
Has the addition of hydrogen effect on the methanogenic community?
Approach:
Development of a new Taqman® assay to study the effects of hydrogen supply on the microbial community
Microbial-DNA isolation from
bioreactor samples
• UltraClean microbial DNA isolation Kit (DNAeasy,
Qiagen)
• Sampling @ different time points
• Filter samples
• ‘Control’ vs in situ or ex situ
• Addition of Internal PCR Control (IPC) primers
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Experimental design: Taqman ®
Internal Control Hydrogenotrophic Acetoclastic Ratio Data Analysis MC = Methanoculleus (H) MSL= Methanosarcinales (A)
Day = 0 Day = 28 1 2 3 MC = Methanoculleus (H) MSL= Methanosarcinales (A)
1. Is the initial situation equal?
2. Does the community change in time? 3. Does the community change after H2
addition? 2 2 2 Day = 21 control in situ Ratio: MC/MSL 2,0 2,7 1,4 40,1 3,5 48,7
Preliminary result:
Clear increase in the presence of hydrogenotrophic methanogens relative to the acetoclastic methanogens as a result of hydrogen supply
Conclusions
•
Hydrogen can be supplied by using diffuse rubber piping in an existing biogas reactor•
Adding hydrogen this way results in more methane per unit biomass•
Hydrogen supply resulted in an increase in the numbers of hydrogenotrophic methanogensFuture plans
• Sequencing MinIon® (work in progress)
• Optimization and extending Taqman® assay*
• Based on lab scale experiments building a pilot reactor (start september 2017)
• Start PhD september 2018 ‘waste to Fuel‘
#MinIon June 06 2017
*A (high) abundance of microorganisms as deduced from metagenome analysis does not necessarily indicate high transcriptional or metabolic activity, and vice versa
Acknowledgements
• Folkert FABER, Emile APOL, Jan Peter NAP (Co-authors) • Hans Banning (CEO Proces Groningen B.V) ✝
• Petr Pozděna (student Univerzita Pardubice, Czech republic) • Sean de Graaf (Onderzoeksmedewerker Alife)
• Linette Oosting and Martijn Boret (Students ILST Biotechnology) • Tom Baars and Henry Wiersma (Students ILST Bioinformatics) • Jan Pieter Thie (ILST Technician)
