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Engineering HMF oxidoreductase for efficient furan building block production

Gupta, Neha LU (2024) KBTM01 20241
Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
Abstract
As the demand for eco-friendly alternatives to traditional plastics grows, Polyethylene Furanoate (PEF) emerges as a promising solution. Derived from renewable biomass sources, PEF embodies sustainability principles by reducing reliance on finite resources and minimizing environmental impact. PEF is synthesized from 2,5-furandicarboxylic acid (FDCA), a key monomer which upon polymerization has the potential to form a variety of bioplastics. The main challenge faced is the low yield of FDCA. To cope with the increasing demand for biobased alternatives, the FDCA production pathway needs to be engineered. The synthesis of FDCA from 5-Hydroxymethyl-2-furancarboxylic acid (HMFCA), mediated by the HMF oxidoreductase (HMFH) gene presents a... (More)
As the demand for eco-friendly alternatives to traditional plastics grows, Polyethylene Furanoate (PEF) emerges as a promising solution. Derived from renewable biomass sources, PEF embodies sustainability principles by reducing reliance on finite resources and minimizing environmental impact. PEF is synthesized from 2,5-furandicarboxylic acid (FDCA), a key monomer which upon polymerization has the potential to form a variety of bioplastics. The main challenge faced is the low yield of FDCA. To cope with the increasing demand for biobased alternatives, the FDCA production pathway needs to be engineered. The synthesis of FDCA from 5-Hydroxymethyl-2-furancarboxylic acid (HMFCA), mediated by the HMF oxidoreductase (HMFH) gene presents a significant bottleneck. HMFCA successful transformation is critical for the overall process. This study aims to enhance the efficiency of HMFH by employing protein engineering techniques including docking, molecular dynamics simulations, and site-directed mutagenesis. Through mutagenesis, significant findings emerged regarding the Thr550Ser and Met58Pro mutants. The wild type showed higher activity than the mutants. Thr550Ser mutant showed nearly complete oxidation of HMFCA, indicating catalytic activity or substrate binding, but to a lesser extent. On the other hand, the Met58Pro mutant exhibited negligible HMFCA oxidation, suggesting a substantial disruption in catalytic behaviour. These observations underscore the impact of mutations on enzyme activity and substrate specificity, offering insights for enzyme optimization in biotechnological applications. To further improve the enzyme's catalytic efficiency and substrate affinity, additional mutation testing could target the D46G, S204G-Y209V, Y209V and V21C mutations. This research contributes to the advancement of sustainable methods for furan building block production, addressing the pressing need for environmentally friendly alternative to fossil-based plastics. (Less)
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author
Gupta, Neha LU
supervisor
organization
course
KBTM01 20241
year
type
H2 - Master's Degree (Two Years)
subject
keywords
Bioplastics, catalytic activity, enzyme optimization, environmentally friendly alternatives, FDCA, furan building block production, HMF oxidoreductase (HMFH) gene, polyethylene furanoate (PEF), protein engineering, substrate specificity, sustainable methods, biotechnology
language
English
id
9161412
date added to LUP
2024-06-12 09:28:39
date last changed
2024-06-12 09:28:39
@misc{9161412,
  abstract     = {{As the demand for eco-friendly alternatives to traditional plastics grows, Polyethylene Furanoate (PEF) emerges as a promising solution. Derived from renewable biomass sources, PEF embodies sustainability principles by reducing reliance on finite resources and minimizing environmental impact. PEF is synthesized from 2,5-furandicarboxylic acid (FDCA), a key monomer which upon polymerization has the potential to form a variety of bioplastics. The main challenge faced is the low yield of FDCA. To cope with the increasing demand for biobased alternatives, the FDCA production pathway needs to be engineered. The synthesis of FDCA from 5-Hydroxymethyl-2-furancarboxylic acid (HMFCA), mediated by the HMF oxidoreductase (HMFH) gene presents a significant bottleneck. HMFCA successful transformation is critical for the overall process. This study aims to enhance the efficiency of HMFH by employing protein engineering techniques including docking, molecular dynamics simulations, and site-directed mutagenesis. Through mutagenesis, significant findings emerged regarding the Thr550Ser and Met58Pro mutants. The wild type showed higher activity than the mutants. Thr550Ser mutant showed nearly complete oxidation of HMFCA, indicating catalytic activity or substrate binding, but to a lesser extent. On the other hand, the Met58Pro mutant exhibited negligible HMFCA oxidation, suggesting a substantial disruption in catalytic behaviour. These observations underscore the impact of mutations on enzyme activity and substrate specificity, offering insights for enzyme optimization in biotechnological applications. To further improve the enzyme's catalytic efficiency and substrate affinity, additional mutation testing could target the D46G, S204G-Y209V, Y209V and V21C mutations. This research contributes to the advancement of sustainable methods for furan building block production, addressing the pressing need for environmentally friendly alternative to fossil-based plastics.}},
  author       = {{Gupta, Neha}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Engineering HMF oxidoreductase for efficient furan building block production}},
  year         = {{2024}},
}