Enhanced 5-Hydroxymethylfurfural (HMF) Conversion to 2,5-Furandicarboxylic Acid (FDCA): Cloning and Expression of FAD-dependent oxidoreductase (HmfH) in Escherichia coli and Gluconobacter oxydans
(2024) KBTM01 20241Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
- Abstract
- The annual plastic production was over 400 million tonnes in 2023 and it is expected to keep growing over the years. The increasing use of fossil-based materials and problems regarding the environmental impact of plastic waste motivate the transition towards a circular economy. Interest in 5-hydroxymethylfurfural (HMF) has increased due to the possibility of being a biobased building block in the synthesis of analogous polyesters like Polyethylene Furanoate (PEF), which is synthesized from ethylene glycol and 2,5-furan dicarboxylic acid (FDCA).
FDCA is derived from HMF through a sequence of oxidation steps through chemical or biological methods. Different enzymes have been used for this oxidation. However, these are limited to only... (More) - The annual plastic production was over 400 million tonnes in 2023 and it is expected to keep growing over the years. The increasing use of fossil-based materials and problems regarding the environmental impact of plastic waste motivate the transition towards a circular economy. Interest in 5-hydroxymethylfurfural (HMF) has increased due to the possibility of being a biobased building block in the synthesis of analogous polyesters like Polyethylene Furanoate (PEF), which is synthesized from ethylene glycol and 2,5-furan dicarboxylic acid (FDCA).
FDCA is derived from HMF through a sequence of oxidation steps through chemical or biological methods. Different enzymes have been used for this oxidation. However, these are limited to only aldehyde or alcohol oxidation, leading to multi-enzymatic cascades to achieve a complete conversion which can affect the process yield.
This study addresses these limitations by exploring the biotransformation efficiencies of HMF into FDCA using recombinant Gluconobacter oxydans DSM 50049 and Escherichia coli. Notable findings were achieved through the expression of FAD-dependent oxidoreductase (HmfH) in E. coli BL21(DE3) with the fusion of maltose binding protein, achieving an 84% conversion of 2 mg mL-1 HMFCA to FDCA. Additionally, the incorporation of the HmfH enzyme into G. oxydans DSM 50049 showed promising results for HMF and HMFCA oxidation, where a 100% and 90% conversion were achieved, respectively. Studies of the incorporation of a coculture system involving the recombinant strains of E. coli and G. oxydans showed a potential strategy to enhance HMf and HMFCA conversion.
This study provides valuable insights into the optimization of enzyme expression and substrate conversion processes for HMF and HMFCA oxidation. Future investigations could further explore alternative enzyme engineering strategies and optimize process parameters for scale-up bioprocesses. (Less) - Popular Abstract
- As the world struggles with the environmental impacts of the plastic industry, finding sustainable alternatives is crucial. In 2023, over 400 million tonnes of plastic were produced, a number that is only expected to increase. Traditional plastics, derived from fossil fuels, contribute significantly to greenhouse gas emissions, making the shift towards a circular and sustainable economy. One promising solution is the production of biobased plastics, which are made from renewable resources like the biomass of plants.
An important chemical derived from plant biomass is 5-hydroxymethylfurfural (HMF), which is used as a platform to produce valuable compounds. This project explores the potential of HMF as a precursor of 2,5-furandicarboxylic... (More) - As the world struggles with the environmental impacts of the plastic industry, finding sustainable alternatives is crucial. In 2023, over 400 million tonnes of plastic were produced, a number that is only expected to increase. Traditional plastics, derived from fossil fuels, contribute significantly to greenhouse gas emissions, making the shift towards a circular and sustainable economy. One promising solution is the production of biobased plastics, which are made from renewable resources like the biomass of plants.
An important chemical derived from plant biomass is 5-hydroxymethylfurfural (HMF), which is used as a platform to produce valuable compounds. This project explores the potential of HMF as a precursor of 2,5-furandicarboxylic acid (FDCA), a biobased compound that can be used to create more sustainable plastics.
FDCA is particularly promising because it is a fundamental ingredient in producing polyethylene furanoate (PEF), a sustainable alternative to conventional plastics like PET. The process of converting HMF to FDCA involves different reaction steps that can be done by chemical (involving high energy consumption) or biological methods, which can sometimes be inefficient.
The research aimed to improve this conversion process by using genetically modified bacteria. Gluconobacter oxydans is a recognized bacteria used in the biotechnology industry to produce valuable compounds. This bacterium is capable do most of the conversion steps fron HMF to FDCA, except for one intermediate (HMFCA). By modifying G. oxydans and another bacterium, Escherichia coli, it was expected to achieve a full conversion of HMF into FDCA.
The study showed that by using only E. coli, an 84% conversion of HMFCA to FDCA was achieved. On the other hand, using G. oxydans led to complete conversion of HMF, and a 90% conversion of HMFCA to FDCA. The most exciting result of the study was using both bacteria together, which made the process more efficient and showed potential to scale up the production of FDCA.
This research contributes to the field of biotechnology aimed at creating sustainable materials. By enhancing the efficiency of HMF conversion through genetic engineering, the future to an increase in the production of eco-friendly plastics is coming closer, benefiting both society and the environment. Future studies will focus on optimizing these processes and scaling up production to meet global demands. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9159880
- author
- Peña Carranza, Leonardo LU
- supervisor
- organization
- course
- KBTM01 20241
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- biotechnology, gluconobacter oxydans, escherichia coli, bioplastics, 5-Hydroxymethyl furfural (HMF), 2, 5-furandicarboxylic acid (FDCA), protein expression
- language
- English
- id
- 9159880
- date added to LUP
- 2024-06-28 09:09:25
- date last changed
- 2024-06-28 09:09:25
@misc{9159880,
abstract = {{The annual plastic production was over 400 million tonnes in 2023 and it is expected to keep growing over the years. The increasing use of fossil-based materials and problems regarding the environmental impact of plastic waste motivate the transition towards a circular economy. Interest in 5-hydroxymethylfurfural (HMF) has increased due to the possibility of being a biobased building block in the synthesis of analogous polyesters like Polyethylene Furanoate (PEF), which is synthesized from ethylene glycol and 2,5-furan dicarboxylic acid (FDCA).
FDCA is derived from HMF through a sequence of oxidation steps through chemical or biological methods. Different enzymes have been used for this oxidation. However, these are limited to only aldehyde or alcohol oxidation, leading to multi-enzymatic cascades to achieve a complete conversion which can affect the process yield.
This study addresses these limitations by exploring the biotransformation efficiencies of HMF into FDCA using recombinant Gluconobacter oxydans DSM 50049 and Escherichia coli. Notable findings were achieved through the expression of FAD-dependent oxidoreductase (HmfH) in E. coli BL21(DE3) with the fusion of maltose binding protein, achieving an 84% conversion of 2 mg mL-1 HMFCA to FDCA. Additionally, the incorporation of the HmfH enzyme into G. oxydans DSM 50049 showed promising results for HMF and HMFCA oxidation, where a 100% and 90% conversion were achieved, respectively. Studies of the incorporation of a coculture system involving the recombinant strains of E. coli and G. oxydans showed a potential strategy to enhance HMf and HMFCA conversion.
This study provides valuable insights into the optimization of enzyme expression and substrate conversion processes for HMF and HMFCA oxidation. Future investigations could further explore alternative enzyme engineering strategies and optimize process parameters for scale-up bioprocesses.}},
author = {{Peña Carranza, Leonardo}},
language = {{eng}},
note = {{Student Paper}},
title = {{Enhanced 5-Hydroxymethylfurfural (HMF) Conversion to 2,5-Furandicarboxylic Acid (FDCA): Cloning and Expression of FAD-dependent oxidoreductase (HmfH) in Escherichia coli and Gluconobacter oxydans}},
year = {{2024}},
}