Balancing the intracellular redox status in engineered autotrophic E. coli
(2023) KMBM01 20231Applied Microbiology
Biotechnology
- Abstract
- Establishing artificial carbon fixation in Escherichia coli as a model organism can open the door to utilization of carbon dioxide as a sustainable feedstock in industrial bioprocesses. Gleizer et al. (2019) developed an autotrophic E. coli strain which combines native and non-native enzymes to create an artificial Calvin Cycle. One of the native enzymes used is the NADH-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In natural autotrophs, the reduction reaction catalyzed by GAPDH is dependent on NADPH. In this study, I tried to establish integration and expression of an NADPH-dependent GAPDH into E. coli with the final goal of creating an NADPH-dependent Calvin Cycle in autotrophic E. coli. Several strategies were used but... (More)
- Establishing artificial carbon fixation in Escherichia coli as a model organism can open the door to utilization of carbon dioxide as a sustainable feedstock in industrial bioprocesses. Gleizer et al. (2019) developed an autotrophic E. coli strain which combines native and non-native enzymes to create an artificial Calvin Cycle. One of the native enzymes used is the NADH-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In natural autotrophs, the reduction reaction catalyzed by GAPDH is dependent on NADPH. In this study, I tried to establish integration and expression of an NADPH-dependent GAPDH into E. coli with the final goal of creating an NADPH-dependent Calvin Cycle in autotrophic E. coli. Several strategies were used but were unsuccessful in creating the desired strains and it remains open whether changing the cofactor dependency can affect growth of autotrophic E. coli. (Less)
- Popular Abstract
- Wherever we look, we are surrounded by things that are made from fossil fuels – the dish soap in our kitchens, the tires on our cars, the sport leggings we wear for the gym. All contain chemicals which are carbon-based. This carbon mostly comes from oil reservoirs hundreds, if not thousands, of meters under the earth’s surface. But what if we didn’t have to use fossil fuels to make carbon-based compounds?
Our cells (and those of every other organism) use and produce a variety of compounds every millisecond – all carbon-based. And we don’t need to drink old dinosaur oil for that (if you want to, that’s your personal choice, but it doesn’t sound very tasty to me). We get everything we need by eating other organisms or their products. The... (More) - Wherever we look, we are surrounded by things that are made from fossil fuels – the dish soap in our kitchens, the tires on our cars, the sport leggings we wear for the gym. All contain chemicals which are carbon-based. This carbon mostly comes from oil reservoirs hundreds, if not thousands, of meters under the earth’s surface. But what if we didn’t have to use fossil fuels to make carbon-based compounds?
Our cells (and those of every other organism) use and produce a variety of compounds every millisecond – all carbon-based. And we don’t need to drink old dinosaur oil for that (if you want to, that’s your personal choice, but it doesn’t sound very tasty to me). We get everything we need by eating other organisms or their products. The same goes for all other animals, fungi and many microorganisms. But plants and some microorganisms go one step further and use carbon dioxide to make the compounds they need. The gas is all around us and doesn’t run out when the harvest was bad or a boat is stuck and blocks international shipping for weeks. Pretty neat, right?
The good news is that we already know how to use cells like bacteria or yeasts to produce a variety of chemicals without fossil fuels. The bad news is that the majority of those cells cannot use carbon dioxide. They eat sugars which need to come from plants which need to grow somewhere, which need to be fertilized, which need to be processed – it’s just an overall needy process. It would make everyone’s life just a bit easier if the cells could consume carbon dioxide directly. And that’s exactly what researchers have been trying to do and have actually achieved – Eureka!
The researchers looked at how plants fix carbon dioxide and imitated the strategy in the bacterium Escherichia coli. It’s functional, but the bacterium is growing so slowly now that in order to work with it you need a patience which most scientists in my field (including me) simply do not have. My goal was to make it grow faster. I tried switching a genetic element, which is required to make sugars from carbon dioxide, with a better one – a process quite similar to tuning a sports car. I didn’t reach my goal in the given time. And maybe that’s an important lesson too.
The public only hears about scientific breakthroughs. We thought about something, we did something, it worked – Eureka! But just as in life, most of the time things in scientific research don’t work or at least not how we expected them to work. We still learn from those experiences and when eventually a little breakthrough does come, we can celebrate it even more. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9139103
- author
- Bohn, Chantal LU
- supervisor
-
- Ed van Niel LU
- organization
- alternative title
- Balansering av den intracellulära redoxstatusen i metaboliskt konstruerad autotrofisk E. coli
- course
- KMBM01 20231
- year
- 2023
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Calvin Cycle, GAPDH, Escherichia coli, C1 Feedstocks, Applied microbiology
- language
- English
- id
- 9139103
- date added to LUP
- 2023-09-26 10:03:29
- date last changed
- 2023-09-26 10:03:29
@misc{9139103,
abstract = {{Establishing artificial carbon fixation in Escherichia coli as a model organism can open the door to utilization of carbon dioxide as a sustainable feedstock in industrial bioprocesses. Gleizer et al. (2019) developed an autotrophic E. coli strain which combines native and non-native enzymes to create an artificial Calvin Cycle. One of the native enzymes used is the NADH-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In natural autotrophs, the reduction reaction catalyzed by GAPDH is dependent on NADPH. In this study, I tried to establish integration and expression of an NADPH-dependent GAPDH into E. coli with the final goal of creating an NADPH-dependent Calvin Cycle in autotrophic E. coli. Several strategies were used but were unsuccessful in creating the desired strains and it remains open whether changing the cofactor dependency can affect growth of autotrophic E. coli.}},
author = {{Bohn, Chantal}},
language = {{eng}},
note = {{Student Paper}},
title = {{Balancing the intracellular redox status in engineered autotrophic E. coli}},
year = {{2023}},
}