Parameters of biogas production:
the microbial connection Gert J.H.Hofstede, Expertise Centre ALIFE, Institute for Life Sciences and Technology, HanzeUniversity of Applied Sciences, Groningen, the Netherlands
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Parameters of biogas production: the microbial connection
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Dr. Ir. Jan Peter Nap
(lector)
Dr. Folkert Faber
(senior lecturer)
Brian Wouterse
(trainee)
Parameters of biogas production:
the microbial connection
In the Netherlands:
About ~180 biogas installations.
In these installations manure and residual materials are (co-)digested.
0,5 billion (=10
9) m
3 biogas is produced
Green electricity can be generated.
Current situation:
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Parameters of biogas production:
the microbial connection
Current situation:
Relatively limited knowledge on the complex community of microorganisms
The economy of biogas plants does not allow failure (cleaning and restarting)
Fairly conservative enterprise
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Parameters of biogas production:
the microbial connection
Why this research ?
The use of defined digestate of microorganisms targeted towards the feedstock might
speed up biogas production.
Opening the microbial black box is supposed to increase biogas yield and the overall
process efficiency.
The overall objective of this research is to describe the microbial community that
participates in the AD
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Parameters of biogas production:
the microbial connection
Have different biomasses distinct biomethane potentials?
Are different methanogenic communities, responsible for biogas production?
Is the methanogenic community influenced by different environments and
different process parameters?
Research question
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Parameters of biogas production: the microbial connection
Experimental tools
Automatic Methane Potential Test System (AMPTSII) batch
2 Labscale bioreactor (10 l, Infors) continuous
Molecular techniques (qPCR )
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Parameters of biogas production: the microbial connection
AMPTSII-experiments: Reproducibility 3 different digestates 2 types of biomass
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Parameters of biogas production: the microbial connection
General setup: 15 bottles, 1 type of biomass (maize), 1 type of digestate (Proces B.V) , 30 days
Digestate to biomass ratio of 1:1 (based on % Organic Dry Matter [ODM]) was used.
General set up of
AMPTSII assays
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Parameters of biogas production: the microbial connection
Conclusion
The AMPTSII yields reproducible results, both in terms of total
gas volume and in terms of methane production
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Parameters of biogas production: the microbial connection
Conclusion:
No significant difference between total biogas productions from
maize by different types of digestates ( Donderen, Lutjegast en
Tolbert).
(ANOVA, one-tail, p=0.05).
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Parameters of biogas production: the microbial connection
Future-plans:
In future experiments, we will look the biogas flow-rate with for
example ‘biomass adapted digestates’ the digestate (Proces
B.V) was not adapted to maize.
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Parameters of biogas production: the microbial connection
The overall objective of this research is to determine the relative changes in
microbial community that participates in the anaerobic digestion on a
Labscale Infors bioreactor
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Parameters of biogas production: the microbial connection
Methanogens, the biological generators of methane
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Parameters of biogas production: the microbial connection
FastRNA® SPIN KIT for SOIL
Total DNA and RNA isolation
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Parameters of biogas production: the microbial connection
Primer combinations used in this study for monitoring the population
dynamics of methanogens in biogas plants
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Parameters of biogas production: the microbial connection
This study revealed the limitation of the qPCR due to the primers. in silico by
the RDP-site
Experimental circumstances probably influence the ‘reference’ Archaea
population
Conclusion/discussion
Commonly used primers in literature should be reconsidered in the light of
expanding data in RDP
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Parameters of biogas production: the microbial connection
Since available primers may not be applicable to all samples,there is a
requirement to design more primers to ensure adequate coverage and
amplification of methanogens.
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Conclusion/discussion
Parameters of biogas production: the microbial connection
Therefore, we also look to at another approach: mRNA of (key-) enzymes that
play a role in the methanogenesis :
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Together with Marina Gil Lopez from the Universitat de Barcelona we did the
first experiments on RNA
Parameters of biogas production: the microbial connection
Identification of Enzymes Involved in synthesis of biogas
The challenge is to find (key-)enzymes that can be linked to (specific
methanogenic) micro-organisms.
as key enzymes can be linked to micro-organisms, relative amounts of micro
organisms can be calculated.
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Parameters of biogas production: the microbial connection
The future for me? I will end up with a bucket full of digestate (or micro-
organisms) optimized for every biomass. That is applied science in my
opinion
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