Optimizing operational parameters for the enzymatic production of furandicarboxylic acid building block

Autor: Ángel T. Martínez, Ana Serrano, María Isabel Sánchez-Ruiz
Přispěvatelé: European Commission, Ministerio de Ciencia e Innovación (España), Consejo Superior de Investigaciones Científicas (España), Martínez, Ángel T. [0000-0002-1584-2863], Serrano, Ana [0000-0002-7057-0418], Martínez, Ángel T., Serrano, Ana
Rok vydání: 2021
Předmět:
5‑Formylfurancarboxylic acid intermediate
Bioengineering
Reaction intermediate
2
5-Furandicarboxylic acid product
010402 general chemistry
Microbiology
01 natural sciences
Applied Microbiology and Biotechnology
Peroxide
Enzyme engineering
Enzyme catalysis
Methylophilaceae
Enzymatic catalysis
chemistry.chemical_compound
Hydrogen peroxide by-product
Dicarboxylic Acids
Hydroxymethylfurfural oxidase
Hydrogen peroxide
Furans
Terephthalic acid
biology
010405 organic chemistry
2
5‑Furandicarboxylic acid product
Research
Reaction pH
Substrate (chemistry)
Catalase
Combinatorial chemistry
QR1-502
0104 chemical sciences
Enzyme inhibition
chemistry
Reaction optimization
biology.protein
5-Formylfurancarboxylic acid intermediate
Oxidoreductases
Oxidation-Reduction
Hydroxymethylfurfural
Biotechnology
Hydrogen peroxide by‑product
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
Microbial Cell Factories
Microbial Cell Factories, Vol 20, Iss 1, Pp 1-13 (2021)
Popis: 13 p.-6 fig.-4 tab.
Background: 2,5-Furandicarboxylic acid (FDCA) is a precursor for green plastics due to its structural similarity to terephthalic acid, a common precursor of oil-derived polymers, and its potential production from sugars obtained from plant biomass. Hydroxymethylfurfural oxidase (HMFO) has been reported as a promising biocatalyst for FDCA production since it can convert bio-based 5-hydroxymethylfurfural (HMF) into FDCA building block. This three-step oxidation reaction occurs through the diformylfuran and 2,5-formylfurancarboxylic acid (FFCA) intermediates. Several efforts have been made for the development of HMFO variants that increase FDCA yields by improving their activities over the reaction intermediates. However, there is still limited insight into how operational conditions can influence these enzymatic reactions. The setup of optimal reaction conditions would enable to understand potential problems hampering the effective industrial production of this bioplastic precursor using HMFO as biocatalyst.
Results: In this work, several parameters affecting the performance of Methylovorus sp HMFO oxidizing HMF have been analyzed for the wild-type enzyme, and its V367R and W466F single variants, V367R/W466F double variant, and I73V/H74Y/G356H/V367R/T414K/A419Y/A435E/W466F (8BxHMFO) octuple variant. Our results show how the oxidation of HMF by HMFO enzymes is highly influenced by pH, with different optimal pH values for the different improved variants. Moreover, the enzymes are not stable at high hydrogen peroxide concentrations and their activity is inhibited by the FFCA intermediate in a pH-dependent way. These limitations can be efficiently overcome with the addition of catalase to the reaction medium, which removes the hydrogen peroxide formed during the oxidations, and the controlled dosage of the substrate to limit the amount of FFCA accumulated in the reaction. The different behavior of wild-type HMFO and its variants against pH, hydrogen peroxide and FFCA highlights the importance of considering each variant as an individual enzyme with its own operational conditions for an eventual industrial FDCA production.
Conclusions: This work provides information of those parameters that condition a high production of FDCA by HMFO. Unraveling these factors allowed to increase the FDCA yields by using the most stable enzymes at their optimal pH for HMF oxidation, removing the peroxide with catalase, and avoiding FFCA accumulation by controlling substrate and/or enzyme concentration. These above findings will be useful when planning a future scale-up of these conversions and will provide new viewpoints for the design of HMFO variants that render a more effective performance during HMF conversion into FDCA.
This work has been funded by the H2020 BBI-JU (https://www.bbi-europe.eu) EnzOx2 (H2020-BBI-PPP-2015-2-720297; https://www.enzox2.eu) project, the GENOBIOREF (BIO2017-86559-R) project of the Spanish Ministry of Science & Innovation (co-financed by FEDER funds), the CSIC project PIE-201620E081, and the CSIC interdisciplinary platform for “Sustainable Plastics towards a Circular Economy” (SUSPLAST).
Databáze: OpenAIRE
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