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QLK3-2000-01528
Biodiversity of methanotrophs and their bioremediation and biotechnological exploitation (BOMBBE)
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Contacts
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Biological Conversion
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Bulk Chemicals
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Quality of Life - 3. The Cell Factory
| Contract No: |
QLK3-2000-01528 |
| Project Type: |
RS (Research and Technological Development Project)
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| Start Date: |
01-10-2000
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| Duration: |
36 months
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| Total Cost: |
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| EC Contribution: |
933 920 EUR
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| Scientific Officer: |
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Abstract
Methanotrophic bacteria can grow in large scale on cheap substrates such as methane
and methanol. They contain unique metabolically versatile enzymes offering enormous potential
for clean biotechnology. However, organisms currently in culture are not sufficiently robust to clean
up organic pollution in bioremediation processes or versatile enough for the production of fine
chemicals. We propose to examine the biodiversity of methanotrophs in more extreme/unexplored
environments. We will use conventional and state-of-the-art molecular ecology techniques to
isolate/characterise novel psychrophilic, thermotolerant, acidophilic, alkaliphilic salt and solvent
resistant methanotrophs containing catalytically versatile methane monooxygenase creating a portfolio
of bacteria for exploitation in biotransformations and bioremediation processes. We will also modify
methanotrophs by mutation/metabolic engineering to create new versatile biological catalysts based
on methane monooxygenase.
Objectives
- To examine the biodiversity of methane oxidising bacteria (methanotrophs) in more
extreme environments.
- To isolate and characterise novel methanotrophs that are able to grow at more extreme
temperatures, pH ranges and that are resistant to salt and solvent stress and can grow in low
nutrient environments.
- To create a portfolio of novel methanotrophs which can be exploited as cell factories in
biotechnology.
- To investigate the potential of these bacteria in bioremediation and biotransformation
processes and for the production of bulk chemicals using clean technology and cheap substrates.
- To improve existing strains of methanotrophs for biotransformation processes by metabolic
engineering.
Description of the work
- A portfolio of novel methane oxidising bacteria will be isolated from a wide range
of environments, including extremes of pH, temperature and salinity by conventional and new
molecular ecological techniques based on isotope probing, the polymerase chain reaction (PCR),
denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridisation (FISH). New
strains will include psychrophiles, thermophiles, alkalophiles and acidophiles and salt and
solvent-resistant strains, more robust than existing methanotrophs with wider applicability
in biotechnology.
- A detailed characterisation of novel methanotrophs at the physiological, biochemical and
molecular level will be carried out to assess their abilities to carry out key co-metabolic processes
under a wide range of growth conditions more relevant for bioremediation and biotransformation
processes. They will also be screened for the production of potentially useful biopolymers and
secondary metabolites.
- Laboratory culture, bioreactor and mesocosm studies will be used to assess the applicability
of new strains in commercially viable biotransformation/co-oxidation reactions and in bioremediation
processes.
- Existing strains of methanotrophs will be improved by metabolic engineering in order to improve
the catalytic utility and performance of these strains in key bioremediation processes for the production
of important metabolites. For example, construction of strains with constitutive production of a
thermostable broad substrate specificity enzyme, soluble methane monooxygenase, in thermophilic
methanotrophs will be carried out. Key strains will also be improved by cloning into them the
bacterial haemoglobin gene to improve their performance in the microaerophilic conditions
encountered in large-scale bioreactors and in field situations during bioremediation
processes.
Deliverables
- Knowledge of the biodiversity of methanotrophs in more extreme environments.
- A culture collection of novel and versatile methanotrophs, highly active in bioremediation
and biotransformation processes which are resistant to more extreme growth conditions in the
environment.
- Exploitation of thermophilic methanotrophs for the production of bulk chemicals from
methane.
- Improved methanotroph strains containing bacterial haemoglobin which grow better
under microaerophilic conditions.
Contacts
Coordinator
Participant
